Solanaceae

Nicotiana rustica

Bibliography

1. C. Abdul Jaleel, G. M. A. Lakshmanan, M. Gomathinayagam, and R. Panneerselvam, “Triadimefon Induced Salt Stress Tolerance in Withania Somnifera and Its Relationship to Antioxidant Defense System,” South African Journal of Botany, vol. 74, no. 1, pp. 126–132, Jan. 2008. doi: 10.1016/j.sajb.2007.10.003. Abstract
   The mitigative effects of triadimefon (5 mg/L) on the germination, early seedling growth, photosynthetic pigments, non-enzymatic antioxidant contents and activities of antioxidant enzymes were studied in salt stressed (40 mM NaCl) Withania somnifera Dunal plants. Salinity stress decreased the germination percentage, early seedling growth and chlorophyll contents. Similarly it affected severely the antioxidants like ascorbic acid (AA), reduced glutathione (GSH) and α-tocopherol (α-toc) in all plant parts (root, stem and leaf). Moreover, it caused changes in the activities of antioxidant enzymes like superoxide dismutase (SOD), peroxidase (POX), polyphenol oxidase (PPO) and catalase (CAT). Triadimefon partially mitigated the salinity stress by enhancing all studied parameters to a considerable extent.
2. P. C. Abhilash and N. Singh, “Withania Somnifera Dunal-Mediated Dissipation of Lindane from Simulated Soil: Implications for Rhizoremediation of Contaminated Soil,” Journal of Soils and Sediments, vol. 10, no. 2, pp. 272–282, Mar. 2010. doi: 10.1007/s11368-009-0085-x. Abstract
   Lindane is an organochlorine chemical that has been used both as an agricultural insecticide and as a treatment for head lice and scabies. It is a neurotoxin that interferes with GABA neurotransmitter function. In humans, lindane primarily affects the nervous system, liver, and kidneys and may be a carcinogen and/or endocrine disruptor. Currently, India is the largest consumer and producer of lindane in the world. Due to its continuous use and indiscriminate industrial production, lindane-contaminated soils are widespread in the country. Apart from India, historical lindane production sites were found in Austria, France, Spain, Bulgaria and in China, Turkey, and the former USSR. Before 1984, lindane was also manufactured in the German Democratic Republic, Poland, Yugoslavia, Romania, and Hungary; since then, all production has been stopped in Germany, Japan, The Netherlands, the UK, and the USA. Because of its worldwide use for more than 50 years, lindane-contaminated soils can be found in most countries of the world. Although many countries have restricted or eliminated its usage, obsolete stock piles continue to pose a threat to various ecosystems and human health. Physical, chemical, and biological methods can all be used for the remediation of contaminated sites, but phytoremediation is now recognized as a cost-effective method for the decontamination of soil sites. The present study examines the potential of Withania somnifera Dunal (previously shown to accumulate lindane from contaminated industrial area; Abhilash et al., Chemosphere 72:79–86, 2008) to take up lindane (γ-HCH) and the subsequent plant-mediated dissipation of lindane from an artificially contaminated soil.
3. A. M. Abou-Douh, “New Withanolides and Other Constituents from the Fruit of Withania Somnifera,” Archiv der Pharmazie, vol. 335, no. 6, pp. 267–276, 2002. doi: 10.1002/1521-4184(200208)335:6<267::AID-ARDP267>3.0.CO;2-E. Abstract
   Two new steroidal lactones of the withanolide-type, 5β, 6α, 14α, 17β, 20β-pentahydroxy-1-oxo-20 S, 22R-witha-2, 24-dienolide (1)and 6α, 7α-epoxy-5α, 14α, 17α, 23β-tetrahydroxy-1-oxo-22R-witha-2, 24-dienolide (2), were isolated from the fruit of Withania somnifera (L.) Dunal (Solanaceae) growing in Southern Egypt, together with two known coumarins, scopoletin (3)and aesculetin (4), a known triterpene, viz. β-amyrin (5), and two known phytosterols, viz. stigmasterol (6) and sitosterol (7). The structures of the isolated compounds have been elucidated by spectroscopic methods including UV, IR, 1H-, 13C-NMR, DEPT experiments, HRFAB-, FAB-, and EI-mass spectrometry. The in vitro antimicrobial activity of the new withanolide (1) and the alcoholic fruit extract are discussed.
4. A. Abraham, I. Kirson, E. Glotter, and D. Lavie, “A Chemotaxonomic Study of Withania Somnifera (L.) Dun.,” Phytochemistry, vol. 7, no. 6, pp. 957–962, Jun. 1968. doi: 10.1016/S0031-9422(00)82182-2. Abstract
   Three chemical types of Wittania somnifera (L.) Dun., Solanaceae, each containing different steroidal lactones of the withanolide type, have been found to occur in Israel; they have been called types I, II and III. Morphological differences could not be detected between the three types, although each of them has a definite and separate area of distribution. No qualitative ontogenetic changes in the withanolide content could be observed.
5. S. R. Adhikari and B. Pant, “Induction and Proliferation of in Vitro Mass of Callus of Withania Somnifera (L.) Dunal,” Research in Plant Sciences, p. 5. Abstract
   Withania somnifera (L.) Dunal. commonly known as “Aswhoganda”, belongs to the family Solanaceae is an important medicinal plant and a major source of alkaloids and steroids. Induction of callus was observed from stem explants on Murashige and Skoog (MS) medium supplemented with various concentrations and combinations of growth hormones viz, 6-Benzylaminopurine (BAP), α-Napthalene acetic acid (NAA), 2,4-Dichlorophenoxy acetic acid (2, 4-D) and kinetin. All the hormone supplemented media gave response for callus growth. The callus was compact and yellowish brown color in all culture condition. The best growth of callus was observed in the MS medium supplemented with 0.5 mgL-1BAP + 1.5 mgL-1 NAA; followed by MS medium supplement with 0.5 mgL-1 BAP + 0.5 mgL-1 NAA, 0.5 mgL-1 BAP + 2.0 mgL-1NAA and 1.0 mgL-1 BAP + 1.5 mgL-1NAA at 8 weeks. Mass production of callus might be used for production and isolation of secondary metabolites for medicinal propose in W. somnifera.
6. M. Adil, B. H. Abbasi, and T. Khan, “Interactive Effects of Melatonin and Light on Growth Parameters and Biochemical Markers in Adventitious Roots of Withania Somnifera L.,” Plant Cell, Tissue and Organ Culture (PCTOC), vol. 123, no. 2, pp. 405–412, Nov. 2015. doi: 10.1007/s11240-015-0844-x. Abstract
   Light plays a pertinent role in plant photo-morphogenesis and it is believed to have an impact on the melatonin-induced physiological functions. In the current study, different light regimes were employed with varying levels of melatonin, either singly or in combination with auxins for the growth and development of adventitious roots in Withania somnifera L. It was observed that 600 µM melatonin favored maximum adventitious root induction frequency (58 %) in cultures incubated under continuous dark conditions. However, adequate root growth (number and length of roots) was observed under 16 h light/8 h dark at 600 µM melatonin. Nevertheless, the interactive effect of light and melatonin was found stimulating for profound production of commercially important secondary metabolites. Correlation among growth parameters and biochemical markers was also observed in the current report. Data on total phenolic content and total flavonoid content were found at higher coincidence with each other and with DPPH antioxidant activity. In conclusion, exogenously applied melatonin mimics IAA activity in root growth and regulates well in 16-h light/8-h dark, thereby giving protection to plant system against light stress.
7. M. Adil, B. Haider Abbasi, and I. ul Haq, “Red Light Controlled Callus Morphogenetic Patterns and Secondary Metabolites Production in Withania Somnifera L.,” Biotechnology Reports, vol. 24, p. e00380, Dec. 2019. doi: 10.1016/j.btre.2019.e00380. Abstract
   Withania somnifera L. is an endangered medicinal plant of higher market value. The in vitro callus cultures were established on Murashige and Skoog (MS) media augmented with different plant growth regulators. The MS medium containing 0.5 mg∙L−1 of each TDZ and NAA was found to be optimal for callus formation and growth. Further, callus cultures were raised in different light wavelengths to find the right wavelength carrying the photons for the ideal cell growth of W. somnifera. Among the different wavelengths, red light was best for maximum biomass accumulation in callus culture. However, violet light condition was proven to be favouring the phenols and flavonoids synthesis in the callus cultures. Compared to other wavelengths, red light grown callus extract showed significantly higher content of chlorogenic acid, and withaferin A. This study concludes that red light treatment was optimum for maximum biomass accumulation and anti-oxidant activity in calli of W. somnifera.
8. A. V. Agarwal, P. Gupta, D. Singh, Y. V. Dhar, D. Chandra, and P. K. Trivedi, “Comprehensive Assessment of the Genes Involved in Withanolide Biosynthesis from Withania Somnifera: Chemotype-Specific and Elicitor-Responsive Expression,” Functional & Integrative Genomics, vol. 17, no. 4, pp. 477–490, Jul. 2017. doi: 10.1007/s10142-017-0548-x. Abstract
   Withania somnifera (L.) Dunal (Family, Solanaceae), is among the most valuable medicinal plants used in Ayurveda owing to its rich reservoir of pharmaceutically active secondary metabolites known as withanolides. Withanolides are C28-steroidal lactones having a triterpenoidal metabolic origin synthesised via mevalonate (MVA) pathway and methyl-D-erythritol-4-phosphate (MEP) pathway involving metabolic intermediacy of 24-methylene (C30-terpenoid) cholesterol. Phytochemical studies suggest differences in the content and/or nature of withanolides in different tissues of different chemotypes. Though development of genomic resources has provided information about putative genes encoding enzymes for biosynthesis of intermediate steps of terpenoid backbone, not much is known about their regulation and response to elicitation. In this study, we generated detailed molecular information about genes catalysing key regulatory steps of withanolide biosynthetic pathway. The full-length sequences of genes encoding enzymes for intermediate steps of terpenoid backbone biosynthesis and their paralogs have been characterized for their functional and structural properties as well as phylogeny using bioinformatics approach. The expression analysis suggests that these genes are differentially expressed in different tissues (with maximal expression in young leaf), chemotypes and in response to salicylic acid (SA) and methyl jasmonate (MJ) treatments. Sub-cellular localization studies suggest that both paralogs of sterol ∆-7 reductase (WsDWF5-1 and WsDWF5-2) are localized in the endoplasmic reticulum (ER) thus supporting their indispensible role in withanolide biosynthesis. Comprehensive information developed, in this study, will lead to elucidation of chemotype- as well as tissue-specific withanolide biosynthesis and development of new tools for functional genomics in this important medicinal plant.
9. A. V. Agarwal et al., “Virus-Induced Silencing of Key Genes Leads to Differential Impact on Withanolide Biosynthesis in the Medicinal Plant, Withania Somnifera,” Plant and Cell Physiology, vol. 59, no. 2, pp. 262–274, Feb. 2018. doi: 10.1093/pcp/pcx179.
10. S. K. Ahirwar, K. K. Agrawal, and H. S. Kushwaha, “Growth and Yield of Ashwagandha [Withania Somnifera (L.)] as Influenced by Different Intercropping System in Kymore Plateau of Madhya Pradesh,” International Journal of Current Microbiology and Applied Sciences, vol. 8, no. 04, pp. 513–524, Apr. 2019. doi: 10.20546/ijcmas.2019.804.056.
11. Ajay, P. Ramesh, K. S. Reddy, S. Ramana, and B. Maji, “Effect of Nitrogen and Farm Yard Manure on Physiological Parameters in Ashwagandha (Withania Somnifera) under Vertisol Soil Type,” Indian Journal of Plant Physiology, vol. 10, no. 4, pp. 389–393, Jul. 2005. https://eurekamag.com/research/004/421/004421036.php. Abstract
    Indian Journal of Plant Physiology 10(4): 389-393
12. R. Akhtar and A. Javaid, “Biological Management of Basal Rot of Onion by Trichoderma Harzianum and Withania Somnifera,” Planta Daninha, vol. 36, Mar. 2018. doi: 10.1590/S0100-83582018360100009. Abstract
    ABSTRACT: Onion is attacked by destructive soil-borne fungal plant pathogen Fusarium oxysporum f. sp. cepae, resulting in basal rot disease. In the present study, three Trichoderma species (T. pseudokoningii, T. harzianum and T. reesei) and leaves of solanaceous weed Withania somnifera were used for management of this disease. The in vitro interaction study revealed T. harzianum as the most effective biocontrol agent against the pathogen. In a pot trial, dried leaf material of W. somnifera (1%, 2% and 3% w/w) and inoculum of T. harzianum were mixed in the pot soil previously inoculated with the pathogen. The highest incidence of the disease (87%) was found in positive control (pathogen inoculation without any amendment). Different rates of dry leaf material reduced the incidence of the disease to 41-66%. T. harzianum in combination with leaf material reduced the incidence of the disease to 20-53%. In a laboratory bioassay, the dry leaf extract of W. somnifera was prepared in methanol and partitioned with n-hexane, chloroform, ethyl acetate and n-butanol. The highest concentration (200 mg mL-1) of all except for the n-butanol fraction significantly decreased fungal biomass over control. This study concludes that basal rot of onion can be controlled by combined application of W. somnifera dry leaf material and biological control agent T. harzianum.
13. N. Akın and İ. Duman, “Improvement of tobacco (Nicotiana tabacum L.) seeds germination properties.,” Ege Üniversitesi Ziraat Fakültesi Dergisi, vol. 55, no. 3, pp. 327–334, 2018. https://www.cabdirect.org/cabdirect/abstract/20183361566. Abstract
    Objective: In the study, provide rapid and homogenous germination of tobacco seeds which are difficult and irregular. Material ve Methods: Varieties of Birlik-125 and Özbaș, tobacco seeds has been used in the study. Effective material, dosage, time and temperature has been determined for improve performance of tobacco germination. For this purpose, were carried out for -6, -12 and -18 MPa doses...
14. H. Al-Ahmad, “In Vitro Decoated Seed Germination and Seedling Development for Propagation of Wild Mandrake (Mandragora Autumnalis Bertol.),” Plants, vol. 9, no. 10, p. 1339, Oct. 2020. doi: 10.3390/plants9101339. Abstract
    The establishment of an efficient in vitro propagation system for the conservation of the Mediterranean Mandragora autumnalis is highly desirable due to its scarcity, besides its potential medicinal and pharmacological properties. In a separate unpublished study, this species has proved to be resistant to laboratory plant regeneration from vegetative tissue cultures; therefore, an alternative decoated seed (i.e., endosperm enclosed the zygotic embryo) germination approach was conducted in this study. Pre-cold treatment of M. autumnalis seeds, removal of seed coats, and exogenous application of gibberellic acid (GA3) promoted in vitro seed germination and seedling emergence. In two separate experiments, approximately 10–27% of the germinated decoated seeds developed healthy seedlings within two weeks, compared to the non-germinated intact seeds of the potting soil controls. After 72 days, the highest rates of healthy seedlings development (67.4 and 69.4%) achieved in the in vitro decoated seed cultures supplemented with 60 and 100 mg/L GA3, respectively, compared to only 25% seedlings emergence rate of the in vitro cultures devoid of GA3, and 44.2% of the soil controls. The in vitro developed plants were healthy, survived transplantation conditions, and, significantly, grew faster, formed on average more than the double number of true leaves and shoot fresh weight (p ≤ 0.05), 90% more fresh weight of root system (p ≤ 0.05), and ultimately more than the double gross fresh weight (p ≤ 0.05) than that of the in vivo developed plants of the soil controls. Such in vitro seed germination approaches would be favorable due to the higher capacity of uniform seedling establishment year-round under lab-controlled conditions, facilitating proliferation and conservation of rare and threatened species, and providing fresh and axenic plant materials required for downstream studies such as those associated with leaf-derived protoplasts and genetic transformations.
15. M. K. S. Al-Banna and J. L. Jinks, “Indirect Selection for Environmental Sensitivity in Nicotiana Rustica,” Heredity, vol. 52, no. 2, pp. 297–301, Apr. 1984. doi: 10.1038/hdy.1984.31. Abstract
    Indirect selection for high and low sensitivity to macroenvironmental differences for final height (FH) has been practised among F2 individuals of the cross of varieties 2 and 12 of Nicotiana rustica using a selection index (R7) proposed by Boughey and Jinks (1978), which is the proportion of FH achieved by the middle of the growing season. The F3 progenies of the selections and of unselected controls were assessed in the range of environments in which the selection index was both measured and originally validated (2 sowing dates ×2 planting densities at the University site), as well as in a similar set of environments at the poorer Avoncroft site. A correlation analysis combined with a significant response to selection confirms that pattern of growth as summarised by R7 is a major determinant of environmental sensitivity of FH to the environmental treatments at the University site and that it has a high repeatability and heritability. The same analysis and the absence of a response to selection at the Avoncroft site show that the usefulness of the selection index is limited to the University site.
16. E. H. A. Al-Doghachi, A.-R. O. Hassan, and N. N. Faris, “Response of Ashwagandha (Withania Somnifera L.) to Sowing Dates and Organic Liquid Fertilizer (Vit-ORG),” Asian Journal of Plant Science and Research, vol. 6, no. 1, pp. 1–7, 2016. Abstract
    The experiment was conducted to study the response of Ashwagandha to sowing dates and organic liquid fertilizer ( Vit-ORG) on chemical components of Ashwagandha sown at Medicinal and Aromatic Plants fields of the College of Agriculture , Basra University ,Iraq, during the growing season of 2013/2014 and 2014/2015. The experiment was laid out in Randomized Complete Block Design in a factorial experiment having three replications. The experiment involved two factors: sowing dates at three levels of 1 or 20 September and 10 October and organic liquid fertilizer ( Vit-ORG) (0 and 2) ml.l-1.Theresults showed that the nutritional contents were significantly affected by sowing dates and organic liquid fertilizer ( Vit-ORG).Percentage of fixed oil and total alkaloid were not significantly affected by sowing dates during both growing seasons. The organic liquid fertilizer ( Vit-ORG) caused increase the percentage of fixed oil in leaves and roots. The interactions between sowing dates and organic liquid fertilizer ( VitORG)gave a significant increase in most studied parameters during both seasons.
17. J. A. Aleksoski and A. T. Korubin-Aleksoska, “Degree of Inheritance and Heritability of Yield in Parental Genotypes and F1 Hybrids of Tobacco,” Journal of Agricultural Sciences, Belgrade, vol. 56, no. 3, pp. 165–172, 2011. doi: 10.2298/JAS1103165A. Abstract
    The mode and level of inheritance of green and dry mass yield per stalk were investigated in four parental genotypes (Burley - B 2/93, Suchum - S1, Suchum - S2 and Prilep - P-84) and in their six diallel F1 hybrids. The trial was set up in 2007, 2008 and 2009 in the field of Tobacco Institute-Prilep in a randomized block design with four replications. The aim of the investigation was to estimate the heritability as an indicator of the inheritance of the yield as one of the most important quantitative characters of tobacco, in order to give suggestions for the selection of parental genotypes and directions for the creation of new varieties. The mode of inheritance was estimated according to the test - significance of the mean value of F1 progeny compared to the parental average. Narrow-sense heritability was estimated after Allard (1960), while broad-sense heritability and genetic components were estimated after Mather and Jinks (1974). The mode of inheritance in the hybrids was different. Positive heterosis for green and dry mass yields per stalk was recorded in S1 x S2. Negative heterosis for green mass yields per stalk was recorded in S1 x P-84 and S2 x P-84, while for dry mass yield it was recorded in S1 x P-84. Inheritance of the characters during the three years of investigation was identical. The higher heritability index of both types was recorded for dry mass yield. As regards inheritance of the yield, the values of broad-sense heritability were higher than those of narrow-sense heritability.
18. J. Ali Lakho, A. Taj, N. Ali Rind, and S. Habib Ahmed Naqvi, “Phytochemical Analysis of <I>Salvadora Oleoides</I> and <I>Withania Somnifera</I>: An Insight into Their Antioxidant and Antimicrobial Capabilities,” Journal of Drug Design and Medicinal Chemistry, vol. 7, no. 1, p. 5, 2021. doi: 10.11648/j.jddmc.20210701.12. Abstract
    Salvadora oleoides and Withania somnifera are the commonly found plants of Pakistan that are known to possess various medicinal properties. Present study was designed to evaluate the presence of various phytochemicals in these plants both qualitatively and quantitatively. This study was concluded with accessing the antioxidant and antimicrobial potentials of these phytochemicals against E. coli, Shigella Spp, Aspergillus terreus and Aspergillus niger. In addition qualitative analysis of phytochemical constituents i.e. alkaloids, glycosides, flavonoids, saponins, steroids, tannins and terpanoids and quantitative analysis of total proteins, sugars, reducing sugars, phenolics and flavonoids was performed. Plant extracts were also checked for the existence of antioxidant and antimicrobial activities. Results revealed the presence of a wide range of phytoconstituents including alkaloids, glycosides, flavonoids, saponins, steroids, tannins and terpenoids in both the plants. Quantification of few pharmaceutically important phytoconstituents showed the diversified response. Presence of antioxidants was confirmed in S. oleoides (2.14±SD mg/mL) in methanol extracts of leaves and water extract of leaves of W. somnifera (1.97±SD mg/mL). Methanolic extracts of leaves, bark and roots of both the medicinal plants showed inhibitory effects against both fungal and bacterial strains used. Our findings provide strong evidence that these medicinal plants possess phytoconstituents of pharmaceutical importance and thus may serve as an effective alternative to routine therapeutics.
19. N. W. Ali, S. Abouzid, A. Nasib, S. Khan, J. Qureshi, and M. I. Choudhary, “RP-HPLC Analysis of Withanolides in the Flowers, Leaves, and Roots of Withania Somnifera,” Acta Chromatographica, vol. 22, no. 3, pp. 473–480, Sep. 2010. doi: 10.1556/achrom.22.2010.3.10. Abstract
    Summary Ashwaghanda, Withania somnifera, is one of the most widely used herbs in Ayurvedic medicine. Leaves and roots are the traditionally used parts of the plant. An RP-HPLC method using a binary acetonitrile-water gradient containing 0.1% acetic acid has been developed for analysis of withaferin A. The method was validated in accordance with ICH guidelines and used for analysis of the withanolide content of the flowers, leaves, and roots of W. somnifera. The withanolide content was highest in the flowers.
20. M. Ali, M. Shuaib, and S. H. Ansari, “Withanolides from the Stem Bark of Withania Somnifera,” Phytochemistry, vol. 44, no. 6, pp. 1163–1168, Mar. 1997. doi: 10.1016/S0031-9422(96)00656-5. Abstract
    Phytochemical studies on the stem bark of Withania somnifera, collected from the southern region of New Delhi, resulted in the isolation of five new withanolides, namely withasomnilide, withasomniferanolide, somniferanolide, somniferawithanolide and somniwithanolide. Their structures have been established as (20R, 22R)-1-oxo-5α, 8β-dihydroxywitha-6α, 7β-epoxide-2,24-dienolide, (20R, 22R)-1-oxo-8β, 11β, 16β-trihydroxywitha-2,5,24-trienolide, (20R, 22R)-1-oxo-8β, 11β-dihydroxywitha-16α, 17α-epoxy-2,5,24-trienolide, (20R, 22R)-1-oxo-8α,18,20β-trihydroxywitha-2,5,24-trienolide and (20R, 22R)-1-oxo-7β, 18,20β,27-tetrahydroxywitha-2,4,24-trienolide, respectively, on the basis of spectroscopic techniques and chemical means.
21. I. Alvarez-López, O. Llanes-Santiago, and J. L. Verdegay, “Drying Process of Tobacco Leaves by Using a Fuzzy Controller,” Fuzzy Sets and Systems, vol. 150, no. 3, pp. 493–506, Mar. 2005. doi: 10.1016/j.fss.2004.07.019. Abstract
    This paper presents the first results from a research project devoted to the application of the fuzzy control based technology to the drying process of tobacco leaves, which the authors are carrying out in the province of Pinar del Rio (Cuba). The fuzzy controller here reported has been designed and implemented by means of a fuzzy rule base constructed from the knowledge of the expert curers taking part in the natural drying process. The preliminary results obtained are very promising.
22. H. M. Alwadi and Z. A. M. Baka, “Microorganisms Asssociated with Withania Somnifera Leaves,” Microbiological Research, vol. 156, no. 4, pp. 303–309, Jan. 2001. doi: 10.1078/0944-5013-00094. Abstract
    Microorganisms including bacteria, actinomycetes and fungi were recovered from the leaves of Withania somnifera, which were collected from two altitudinal ranges (0–300 m and 1700–2000 m) in the Asir region, Saudi Arabia. Types and numbers of microorganisms varied according to the altitude and the month of collection. The number of microorganisms was higher on old leaves than that on young ones in most cases. Low altitude exhibited more microorganisms than high altitude. The relationship between meteorological factors and type and number of the recovered microorganisms is discussed. Inoculation of detached healthy leaves of Withania by all recovered fungal species revealed only Alternaria solani as a pathogen of this plant. To confirm pathogenicity, scanning and transmission electron microscopic examination revealed the colonization of this pathogen inside the leaf tissue. Penetration of Withania leaves by the fungus occurred only through stomata, and the invading hyphae were located in the intercellular spaces of leaf tissues. Ultrastructural changes noted in infected cells included changes in chloroplasts and the invagination of the host plasma membrane.
23. T. M. Anandakumar, D. Kumar, B. Shivanna, and R. Kumar, “Physical Properties of Ashwagandha Seeds (Withania Somnifera L.) – A Medicinal Crop,” Industrial Crops and Products, vol. 186, p. 115233, Oct. 2022. doi: 10.1016/j.indcrop.2022.115233. Abstract
    Ashwagandha (Withania somnifera L.) is a potential medicinal plant belongs to the Solanaceae family, offers various health benefits, such as enhancement of memory, improved blood sugar, and inhibition of inflammation, stress and anxiety, and boosted in muscle strength and fertility. Hence, the demand for the roots of ashwagandha is very high in the Indian market, thus the ashwagandha crop needs to be cultivated in a larger area to mitigate the demand of roots in Indian as well as foreign markets. To cultivate this crop in larger areas with less time and manpower consumption, mechanization of sowing operation is very much essential. In this regard, this study has been conducted to determine the physical properties of ashwagandha seeds (Five cultivars, namely NMITLI-101, NMITLI-118, CIM-Pushti, Pratap and Poshitha) for designing and developing of seed sowing equipment. The data collected were analyzed on R software based web application developed by department of agricultural statistics, college of agriculture, Vellayani, Keral Agricultural University, Vellanikkara, Kerala. Results revealed that cv. CIM-Pushti seed has significantly higher values as compared with other cultivars in major, minor, intermediate diameter, arithmetic mean diameter, and geometric mean diameters. Similarly, in the case of physical properties like sphericity, surface area, sample volume, and elongation ratio, cv. CIM-Pushti showed significantly higher values as compared with other cultivars whereas in the case of aspect ratio and flakiness ratio, cv. Poshita showed significantly higher values. In the case of gravimetric properties, cv. Pratap and NMITLI-118 resulted in higher values of test weight (1000 seeds weight) as compared with other cultivars. Further, cv. Pratap and CIM-Pushti have significantly higher values of bulk density and true density over other cultivars, respectively. These results are likely to be very much helpful for agricultural engineers to design various seed handling tools/machines like conveying machines, storage containers, processing machines and sowing equipment for Ashwagandha seeds.
24. R. A. Andersen, H. R. Burton, P. D. Fleming, T. R. Hamilton-Kemp, and S. L. Gay, “Effects of Air-Curing Environment on Alkaloid-Derived Nitrosamines in Burley Tobacco,” IARC scientific publications, no. 84, pp. 451–455, Jan. 1987. Abstract
    Levels of nitrite and pyridine alkaloid-derived total tobacco-specific nitrosamines (TSNA) were significantly higher in tobacco leaf (normal or late harvest) air-cured at 32 degrees C/83% relative humidity (RH) than in more moderate environments, i.e., 15 degrees C/50% RH and 24 degrees C/70% RH. These constituents increased appreciably from day 10 to day 21 of the cure. The near-concurrent appearances of maximal total contents of TSNA [sum of N’-nitrosonornicotine (NNN), N’-nitrosoanatabine (NAT), N’-nitrosoanabasine (NAB) and 4-(N-nitrosomethylamino)-1-(3-pyridyl)-1-butanone) (NNK)] and nitrite supports the concept that nitrite is a limiting and proximal precursor of total TSNA during the curing of tobacco. During a long curing period (50 days) at 32 degrees C/83% RH, nicotine and anatabine contents decreased, but TSNA contents increased in leaves of all harvest dates and stalk positions. These results support the view that nicotine and anatabine are precursors of TSNA. Measurement of nitrite and individual alkaloids during post-harvest processing of tobacco leaf may provide an index of the potential accumulation of alkaloid-derived nitrosamines.
25. G. J. Anderson, G. Bernardello, M. R. Opel, A. Santos-Guerra, and M. Anderson, “Reproductive Biology of the Dioecious Canary Islands Endemic Withania Aristata (Solanaceae),” American Journal of Botany, vol. 93, no. 9, pp. 1295–1305, 2006. doi: 10.3732/ajb.93.9.1295. Abstract
    We confirmed functional dioecy of Withania aristata via field and greenhouse studies. Male flowers are significantly larger. Female flowers bear stamens with no pollen; males bear 220 000 grains. Stigmata of male flowers senesce in buds. Anatomical observations confirm more ovules in females and an ovarian nectary in both sexes. We detected nectar in female flowers in the greenhouse but found no nectar in males. Thus, males offer pollen and females nectar. Females bear large numbers of fruits and, infrequently, male plants bear few significantly smaller fruits with few seeds. Outcrosses of females (self crosses impossible without pollen) yielded fruits in young buds, older buds, and open flowers. Self crosses of male flowers succeeded only with very young buds. Although functionally dioecious, this species manifests self-compatibility; however, no fruits are produced autonomously. Bee species (Lassioglossum, Amegilla, Apis) visit flowers and mature buds. Bud visits in which bees force petal tips apart, coupled with self-compatibility, may explain infrequent fruit on males. Thus, dioecy in W. aristata seems to have evolved from self-compatible ancestors, that leaky dioecy may have been favored during colonization, and, that despite autogamy and a low floral visition rate, this endemic enjoys a high rate of reproductive success.
26. A. S. R. Anjaneyulyu and D. S. Rao, “A New Withanolide from the Leaves of Withania Somnifera,” Indian Journal of Chemistry, vol. 36B, pp. 161–165, Feb. 1997. Abstract
    From the leaves of Withania sonmifera (Solanaceae ),collected from Tamil Nadu, anew withanolide, 3a~ethoxy-2,3-dihydro-27-deoxywithaferin A(1) inaddition tothe known derivatives withaferin A(4), 2,3dihydrowithaferin A(5) 3p-methoxy-2,3-dihydrowithaferin A(2) and 27-deoxywithaferin A(3) have been isolated, and their structures elucidated onthe basis ofphysical, chemical and spectral methods.
27. G. M. Ankad, J. Hiremath, R. T. Patil, H. J. Pramod, and H. V. Hegde, “Evaluation of Kunapa Jala and Pancha Gavya on Plant Quality, Physiology, Biochemistry, Yield and Growth Attributes – A Case Study of Withania Somnifera Dun.,” Journal of Ayurveda and Integrative Medicine, vol. 8, no. 4, pp. 247–251, Oct. 2017. doi: 10.1016/j.jaim.2017.01.004. Abstract
    Background ‘Vrikshayurveda’, an ancient science of plant life described by Surapala, has clearly outlined a systematized agricultural practice that insisted on the use of Kunapa jala (KJ) and Pancha gavya (PG) to enhance the yield and quality of plants. Objective An experiment was conducted to evaluate the effect of KJ and PG on growth, physiological, biochemical, quality attributes and yield of Withania somnifera Dun. Materials and methods The effect of KJ and PG was evaluated in comparison with control, organic (farmyard manure and humic acid) and inorganic (NPK) fertilizer at 60, 90, 120 and 150 days after sowing (DAS). The study was conducted in randomized complete block design method. Results KJ group were higher in, total leaf area (1707.89 cm2) at 120 DAS, leaf area index (3.795) at 120 DAS, crop growth rate (0.256 g m−2 day−1) at 60–90 DAS, leaf area duration (101.909) at 120–150 DAS, relative growth rate (0.0170 g g−1 day−1) at 60–90 DAS, net assimilation rate (0.0537 g m−2 day−1) at 60–90 DAS. Leaf area ratio was higher in PG group (37.937 m2 g−1) at 60 DAS. The higher levels of chlorophyll a, b and carotenoids were in KJ group (1.877, 0.745 and 1318.14 mg g−1 respectively) at 90 DAS. Yield and quality attributes at harvest (150 DAS) indicated, higher dry root yield (5.93 quintal hectare−1), root length (15.66 cm) were higher in KJ group and root diameter was higher in PG group (1.36 cm). Conclusion Vrikshayurveda practices viz. Kunapa jala and Pancha gavya were effective on studied parameters of W. somnifera. Economical and eco-friendly Vrikhayurveda practices can be initiated for sustainable agriculture.
28. M.-M. Antofie and C. Sava Sand, “Drought Stress Study on Nicotiana Tabacum L., ‘Baladi’, an In Vitro Experimental Model,” Agriculture, vol. 11, no. 9, p. 845, Sep. 2021. doi: 10.3390/agriculture11090845. Abstract
    Crops drought tolerance is a trait of outmost importance for agriculture especially today when climate change is affecting more the production for food and feed. The scope of this article is to evaluate in vitro drought stress response of Nicotiana tabacum L., “Baladi”. The experiment was set up for four successive stages starting with in vitro seedling development, hypocotyl cultivation, three generations of micropropagation, pre-acclimatization and acclimatization. The effect of abscisic acid (ABA) and/or polyethylene-glycol 6000 (PEG) on tobacco hypocotyl caulogenesis and micropropagation were investigated. Superoxide-dismutases (SODs) and peroxidases (POXs) are more active and different isoforms patterns have been identified compared to the control for cualogenesis. A decrease of internodes length and a higher shoots multiplication rate were observed. However, under PEG treatment plantlets expressed hyperhydration and ceased rooting. Pre-treatments effects study of ABA and/or PEG were finalized in acclimatization phase for 18 tobacco clones. A summary of our results revealed that ABA and/or PEG induce among others a higher oxidative stress compared to the control in the first stage that is not maintained for all clones until acclimatization. Certain clones expressed a lower SOD activity compared to the control during acclimatization but maintaining higher POX activity.
29. N. Anuroopa and D. J. Bagyaraj, “Influence of Different AM Fungi on the Growth, Nutrition and Withanolide Concentration of Withania Somnifera,” Medicinal Plants - International Journal of Phytomedicines and Related Industries, vol. 7, no. 4, p. 272, 2015. doi: 10.5958/0975-6892.2015.00040.4.
30. N. Anuroopa, “Inoculation with Selected Microbial Consortia Not Only Enhances Growth and Yield of Withania Somnifera but Also Reduces Fertilizer Application by 25% under Field Conditions,” Proceedings of the Indian National Science Academy, vol. 93, no. INVALID_SCITE_VALUE, Aug. 2017. doi: 10.16943/ptinsa/2017/49127.
31. R. L. Arya, P. H. Kumar, S. Roy, C. Chandrasekhararao, S. Amarnath, and S. Chanda, “Effect of Integrated Nutrient Management on Yield, Quality, and Economics of Motihari I Tobacco (Nicotiana Rustica) in Terai Region of North Bengal,” Jun. 2011. http://krishi.icar.gov.in/jspui/handle/123456789/50097. Abstract
    Tobacco is one of the important cash crops grown particularly in northern part of West Bengal. Two cultivated species of tobacco viz., Nicotiana tabacum and Nicotiana rustica locally known as Jati and Motihari tobacco, respectively are grown in Cooch Behar and Jalpaiguri districts of West Bengal. Tobacco area in West Bengal is 18,000 ha of which about 14,000 ha is under Motihari tobacco which holds traditional reputation for its quality and strong flavour and is used mainly for hookah as well as chewing purposes. In Motihari tobacco, Hemti and Bitri types are characterized by late and early maturing genotypes/land-races, respectively. Judicious use of organic manures with inorganic fertilizers not only increases the yield of tobacco but also makes the tobacco farming sustainable under North Bengal conditions (Krishnamurthy et al., 1990). The phosphate solubilizing bacteria play a significant role in dissolving interlocked phosphates in the soil into available form (Sharma, 2003). The role of Azotobacter in fixing atmospheric nitrogen by using organic carbon and phosphate has been established (Lehri and Mehrotra, 1972; Shinde et al., 1977; Harishu Kumar et al., 1991). Keeping in view the adverse effect of chemical fertilizers on soil and crop health and also on beneficial effects of microorganisms, the present study was conducted to investigate the role of chemical fertilizers along with promising biofertilizers on yield and quality of Motihari tobacco.
32. Asha Rani N.S and Prasad M.P, “In-Vitro Studies on the Germination of Atropa Belladonna Seeds under Different Conditions,” International Journal of Science and Research, vol. 3, no. 10, 2012. https://www.ijsr.net/archive/v3i10/T0NUMTQxODQ=.pdf. Abstract
    Atropa belladonna contains tropane alkaloids and a raw material for pharmaceutics industry. Large scale cultivation of Belladonna is carried both by bulb and seeds. Seeds are usually sown during the first half of March and it take almost 3 months for germination and hence farmers prefer bulbs of belladonna for cultivation. Prolong germination of seed is due to seed Dormancy. Dormancy is a mechanism to prevent germination during unsuitable ecological conditions, when the probability of seedling survival is low. The present investigation was carried out to improve germination percentage of Atropa belladonna seeds by different Scarification methods. Mechanical and Chemical Scarifications were carried on seeds of belladonna. Both mechanical and chemical methods significantly stimulated seed germination in varying percentage. Seeds were germinated on full strength M S media with or without GA3 under invitro condition. Maximum germination was obtained by Acid treatments and also by boiling water treatments. Hard impermeable Testa of the seeds was successfully broken by above Scarification treatment and even insufficient germinating hormones were also supplied for good germination percentage in present investigation.
33. U. S. E. P. A. N. C. for E. Assessment, “Large Scale Production of Withania Somnifera (L.) Dunal Using in Vitro Techniques.” Mar-2009. https://hero.epa.gov/hero/index.cfm/reference/details/reference_id/8450350.
34. U. S. E. P. A. N. C. for E. Assessment, “Novel Method to Isolate Withaferin A from Withania Somnifera Roots and Its Bioactivity.” Mar-2009. https://hero.epa.gov/hero/index.cfm/reference/details/reference_id/7738795.
35. N. Atri et al., “Screening for Endophytic Fungi with Antibacterial Efficiency from Moringa Oleifera and Withania Somnifera,” Journal of scientific research, vol. 64, no. 01, pp. 127–133, 2020. doi: 10.37398/JSR.2020.640118. Abstract
    Plants have a great potential to grow while carrying inside them fungi and bacteria known as endophytes. These endophytes have a potential to produce bioactive molecules which are of immense pharmaceutical importance. In the current study, plants having medicinal properties such as Moringa oleifera and Withania somnifera were explored for studying endophytic fungi associated with them. Moringa oleifera and Withania somnifera are widely known for their nutritional and medicinal values. The endophytic fungal associations of these plants have revealed their efficacy in therapeutic field. In the current study, endophytic fungal strains were extracted by incubating the dried plant samples on potato dextrose agar media, supplemented with streptomycin at 28 °C for 5-10 days. A total of 21 and 24 endophytes were isolated from herbal plant Moringa oleifera and Withania somnifera, respectively. Microscopic study revealed four active strains of M. oleifera i.e. Colletotrichum sp. (MO-S2), Cladosporium sp. (MO-S4, MO-L3) and Fusarium sp. (MO-R1), and two active strains of W. somnifera i.e. Alternaria sp. (WS-S8) and Fusarium sp. (WS-R5). Active strains of endophytic fungi were further screened for their antibacterial activity against pathogenic gram negative bacterial strain of Escherichia coli (ATCC 25922) and gram positive strain of Staphylococcus aureus (ATCC25323). Our results showed that fungal isolates of M. oleifera (MO-S2) and W. somnifera (WS-S8) had antibacterial activity against both the bacterial strains. However, three fungal strains of M. oleifera (MO-L3, MO-S4, and MO-R1) and one of W. somnifera (WS-R5) showed antibacterial activity against gram negative bacterial strain of E. coli.
36. R. H. Autade, S. A. Fargade, A. R. Savant, S. S. Gangurde, R. S. Choudhary, and S. S. Dighe, “Micropropagation of Ashwagandha (Withania Somnifera),” BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS, p. 7. Abstract
    In the present work in vitro propagation of a multipurpose medicinal plant, Withania somnifera was done. Direct regeneration of nodal explants and their multiplication have been optimized using cytokinin BAP (0.5-4.0 mg/l) and combination of BAP (0.5 mg/l) + NAA (0.5-3.0 mg/l) respectively. MS media with nodal explants supplemented with BAP (2.0 mg/l) produced maximum average number of shoots (2±0.37) and average shoot length was found to be 2.8±0.15 cm. Best initiated shoots then sub cultured for shoot multiplication, an improved shoot multiplication in terms of average number of shoots (5.3±0.41) and average shoot length 6.5±0.12 cm was observed on MS media in combination with BAP (0.5 mg/l) + NAA (1.5 mg/l). Maximum average number (12± 0.20) and average length 9.8±0.26cm of roots were observed on MS media supplemented with IBA (2.0 mg/l) out of different IBA (1-5 mg/l) concentrations were taken in to consideration during the study. Regenerated plantlets were successfully transferred to greenhouse condition.
37. M. F. Azhar et al., “Antioxidant and Phytochemical Composition of Leaves, Stem and Root Extracts of Withania Coagulans and Withania Somnifera,” p. 4, 2020. Abstract
    Different plant parts of Withania coagulans and Withania somnifera belonging to family Solanaceae were exploited for biochemical studies. These are wild plants and mostly considered as poisonous plants, although limitedly used in folk medicines in Pakistan. The extraction yield in menthol was also calculated for root, stem and leaves of both plants. The\,% of extract yield in leaves of W. coagulans was recorded high (7.6\,%) than stem (6.3\,%) and root (6.3\,%). The\,% extract yield of all parts of W. coagulans was greater than the same parts of W. somnifera. The pattern of\,% extract yield is similar for W. somnifera as in W. coagulans. In leaves its higher (5.5\,%) and a decreases from stem (5.2\,%) to roots (4.7\,%). Flavonoids and total phenolics were determined by using Folin-Ciocalteu reagent and aluminum chloride reagent through spectrophotometric assay. Maximum value of total phenolics and flavonoids were observed in W. coagulans leaves (58.21 mg GEA/g and 47 mg RE/g). There is a similar decreasing pattern in total phenolics and flavonoids contents from leaves to stem and root extracts. Antioxidant activities in different parts extracts of both plants were calculated at different levels. All parts of W. coagulans exhibits more antioxidant activity than W. somnifera. These results reveal the presence of useful biomedical components in both species which could be used in formulation of modern medicines.
38. G. Baghel, Q. A. Naqvi, S. K. Snehi, M. S. Khan, and S. K. Raj, “Molecular Identification of Three Isolates of Jatropha Mosaic India Virus Associated with Mosaic Disease of Withania Somnifera in India,” Archives of Phytopathology and Plant Protection, vol. 45, no. 17, pp. 2114–2119, Oct. 2012. doi: 10.1080/03235408.2012.721683. Abstract
    The association of begomovirus with mosaic disease of Withania somnifera was detected at three locations in India by PCR using begomovirus-specific degenerate primers. The resulting amplicons of ∼1.2 kb from three locations (Aligarh, Lucknow and Hindaun City) were sequenced. The begomovirus isolates of three locations shared 96–97% identities among them and 91% identities with Jatropha mosaic India virus (JMIV). Based on highest sequence identities and close phylogenetic relationships with JMIV, these begomovirus isolates associated with mosaic disease of W. somnifera were identified as isolates of JMIV.
39. S. Bahrami-Rad and R. Hajiboland, “Effect of Potassium Application in Drought-Stressed Tobacco (Nicotiana Rustica L.) Plants: Comparison of Root with Foliar Application,” Annals of Agricultural Sciences, vol. 62, no. 2, pp. 121–130, Dec. 2017. doi: 10.1016/j.aoas.2017.08.001. Abstract
    Effect of potassium (K) application through leaves (LA) or roots (RA) was studied in tobacco plants grown under K deficiency and drought stress conditions. Application of K was effective in improving the shoot growth only under drought conditions, whereas root biomass and length responded under both watering regimes. Under drought conditions, photosynthesis and transpiration activities increased upon K application leading to a reduced water use efficiency. Both RA and LA increased the leaf water potential, relative water content and turgor under both well-watered and drought conditions; RA was more effective than LA in the recovery of leaf turgor. Analyses of water relation parameters in different aged leaves showed lower susceptibility of the middle-aged leaves to both K deficiency and drought stresses than the upper and lower leaves; this phenomenon was accompanied by a more conservative control of water loss in the middle-aged leaves. In contrast, proline was accumulated in the young leaves, and K application increased it further. Although various organic osmolytes were accumulated under the combinative effect of K deficiency and drought stress, they did not exceed the amounts found in the control (well-watered +K) plants and were merely a result of the concentration effect. Collectively, our results revealed that the majority of leaf biochemical responses to drought stress are developmentally regulated processes. In addition, the alleviating effect of both RA and LA despite higher water loss indicated that an improved stomatal function upon K application allowed carbohydrates synthesis, thus, enhancing plant growth under water stress.
40. A. Baldi, D. Singh, and V. K. Dixit, “Dual Elicitation for Improved Production of Withaferin A by Cell Suspension Cultures of Withania Somnifera,” Applied Biochemistry and Biotechnology, vol. 151, no. 2, p. 556, May 2008. doi: 10.1007/s12010-008-8231-2. Abstract
    High yielding transformed callus culture of W. somnifera was established by infecting hypocotyls with Agrobacterium tumefaciens MTCC-2250. Maximum withaferin A content of 0.0875 mg/g dry cell weight and transformation efficiency of 80% were obtained. Confirmation of transformation was done on the basis of the presence of the ags gene by using polymerase chain reaction. Various abiotic elicitors (arachidonic acid, methyl jasmonate, calcium chloride, and copper sulfate) and biotic elicitors (cell extracts and culture filtrates of Alternia alternata, Fusarium solani, and Verticilium dahaliae) were tested at different concentrations to enhance withaferin A production in suspension culture of transformed cells. Maximum enhancements of 5.4 times and 9.7 times, respectively, were obtained when copper sulfate (100 μM) and the cell extract of V. dahaliae (5% v/v) were added separately to suspension cultures. The dual elicitation strategy by the combined addition of these two elicitors resulted in 13.8-fold enhancement of withaferin A content in comparison to control cultures (2.65 mg/L). The present study indicates the potential of this biotechnology-based methodology for the large-scale production of withaferin A.
41. I. T. Baldwin, “Damage-Induced Alkaloids in Tobacco: Pot-Bound Plants Are Not Inducible,” Journal of Chemical Ecology, vol. 14, no. 4, pp. 1113–1120, Apr. 1988. doi: 10.1007/BF01019339. Abstract
    Field-grown wild tobacco plants (Nicotiana sylvestris) were subjected to a defoliation regime designed to mimic the rate and amount of leaf mass removed by one tobacco hornworm per plant. Undamaged leaves on these plants undergo a dramatic (457% for leaf position 5, 410% for leaf position 8) increase in total leaf alkaloids compared to same-age and positioned control leaves on undamaged control plants. However, potted greenhouse-grown plants fail to exhibit the same damage-induced increase in alkaloid content. The greenhouse environment differs from the field environment in factors known to influence leaf alkaloid content, particularly soil N, P, K, near-UV radiation, and relative humidity. However, altering these environmental factors does not make potted plants able to increase their leaf alkaloid levels in response to defoliation. Transplanting plants into larger pots with more soil does allow the plants to respond to defoliation. Thirty days after transplanting, the plants are again unresponsive to damage, probably as a result of becoming “pot-bound.” This result suggests a mechanism for the induction response, specifically that leaf damage triggers synthesis of these alkaloids in the roots, and offers a potentially valuable experimental tool for the study of induced-plant defenses in tobacco and other plants that synthesize alkaloids in their root tissues.
42. I. T. Baldwin, R. C. Oesch, P. M. Merhige, and K. Hayes, “Damage-Induced Root Nitrogen Metabolism in Nicotiana Sylvestris: Testing C/N Predictions for Alkaloid Production,” Journal of Chemical Ecology, vol. 19, no. 12, pp. 3029–3043, Dec. 1993. doi: 10.1007/BF00980600. Abstract
    “Nitrogen surplus” models for nicotine production induced by leaf damage predict that the observed increase in root nicotine synthesis after leaf damage results from “overflow” metabolism; reduced nitrogen existing in excess of growth requirements is shunted into nicotine biosynthesis. To test the nitrogen surplus model for induced nicotine production, we measured the concentrations of the majorN-containing metabolites exported from the roots and the nitrate reductase activity (NRA) of roots and shoots in damaged and undamagedNicotiana sylvestris plants. Leaf damage: (1) had no significant effect on root or shoot NRA, (2) increased nicotine export and decreased amino-acid and amide export from the roots of NO3-fertilized plants, and (3) had no significant effect on the export of anyN-containing metabolite from the roots of NH4-fertilized plants. These results are not consistent with the nitrogen surplus model and indicate that leaf damage has a direct influence on root alkaloid metabolism.
43. I. T. Baldwin, “An Ecologically Motivated Analysis of Plant-Herbivore Interactions in Native Tobacco,” Plant Physiology, vol. 127, no. 4, pp. 1449–1458, Dec. 2001. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1540177/.
44. I. T. Baldwin, “Inducible Nicotine Production in Native Nicotiana as an Example of Adaptive Phenotypic Plasticity,” Journal of Chemical Ecology, vol. 25, no. 1, pp. 3–30, Jan. 1999. doi: 10.1023/A:1020880931488. Abstract
    Nicotine, an inducible defense in a number of Nicotiana species, exemplifies adaptive phenotypic plasticity. The mechanisms responsible for its production are reviewed, and the induced character states are characterized allometrically in order to understand how inducibility changes over ontogeny responds to environmental variables that influence plant growth, and to relate inducible production to plant fitness correlates. The empirical evidence for fitness costs and benefits of inducible nicotine production are considered, and the physiological and ecological mechanisms potentially responsible for the costs are considered. An intimate understanding of the plant’s natural history is an essential prerequisite to understanding these costs and benefits. Inducible nicotine production is just one of many traits that are altered after herbivore attack, and the cost–benefit model provides a valuable heuristic framework in which to understand the selective factors responsible for the maintenance of inducibly expressed traits.
45. I. T. Baldwin, “Methyl Jasmonate-Induced Nicotine Production in Nicotiana Attenuata: Inducing Defenses in the Field without Wounding,” in Proceedings of the 9th International Symposium on Insect-Plant Relationships, E. Städler, M. Rowell-Rahier, and R. Bauer, Eds. Dordrecht: Springer Netherlands, 1996, pp. 213–220. doi: 10.1007/978-94-009-1720-0_49. Abstract
    The functional significance of herbivore-induced plant traits known to directly or indirectly influence herbivore performance remains largely untested under field conditions due to the difficulty of uncoupling the response to herbivory from the act of herbivory. The signals that activate many of the induced responses in plants are endogenously produced in response to wounding, unlike many of the predator-induced responses found in aquatic invertebrates (which are activated by exogenous cues derived from predators). Jasmonates, endogenously-produced damage signals, activate diverse wound-induced responses in plants including induced nicotine production in Nicotiana sylvestris. The results presented here are from two experiments which illustrate the use of jasmonates to uncouple induced nicotine production in Nicotiana attenuata (Torrey ex. Watson) from wounding. The exogenous addition of methyl jasmonate (MJ) in small quantities (11 βg for a 1.4 g dry mass plant) to roots of hydroponically-grown plants induces de novo nicotine synthesis and increases whole-plant nicotine concentrations just as wounding does. The MJ-induced changes were proportional to the quantity of MJ given. Moreover, the effects of MJ were additive to the effects of damage. Applications of MJ to shoots were less effective. Root treatments also worked with plants growing in a field plot. The application of MJ represents a promising tool for examining the functional significance of induced nicotine responses in plants growing in their native environments.
46. I. T. Baldwin and L. Morse, “Up in Smoke: II. Germination ofNicotiana Attenuata in Response to Smoke-Derived Cues and Nutrients in Burned and Unburned Soils,” Journal of Chemical Ecology, vol. 20, no. 9, pp. 2373–2391, Sep. 1994. doi: 10.1007/BF02033208. Abstract
    Nicotiana attenuata is a native tobacco that is commonly found usually one growing season after fires in the blackbrush, sagebrush and pinyon-juniper forests of the Great Basin desert of North America. This plant also occurs in isolated dry washes and roadsides for many consecutive seasons. Postfire annuals are thought to synchronize their germination from the seed bank with the postfire environment in response to increases in (1) fire-related cues or (2) nutrient supply rates resulting from the mineralization of nutrients by fire, or (3) the removal of allelochemicals produced by the dominant vegetation occupying the sites before the burn or the microbial community associated with the dominant vegetation. We examine the effect of these three changes on the germination ofN. attenuata seed from artificial seed banks made with burned and unburned soil taken in 1993 from under four dominant shrub species (Coleogyne ramosissima, Yucca baccata, Lycium andersonnii, Purshia tridentata) of an area that burned in 1992 and from two dry washes in whichN. attenuata populations have persisted since at least 1988. We utilize our recent discovery that aqueous extracts of wood smoke contain potent germination cue(s) for this species and the established observation that nitrate stimulates germination in manyNicotiana species. In two experiments, we added smoke-derived germination cues and nutrients separately and in combinations to the artificial seed banks, measured germination rates, and inferred the effect of burning by the response of the seed banks to these additions. Germination rates of seed in burned soil were consistently higher than those in unburned soil collected from under all species tested; concentrations of nitrate, P, Mn, and Ca were also higher in burned than unburned soils. Because the addition of more cue and nitrate to burned soil increased germination rates, these soil components may not be at concentrations sufficient to saturate the germination response one year after a fire. The germination of seeds in soil collected from beneath unburnedYucca bacatta plants increased to the same level as that found under burned plants of the same species with the addition of cue and nitrate. Similarly, unburned bitterbrush soil attained the same germination potential with the addition of cue and a complete nutrient solution as burned bitterbrush soil. We conclude that the effect of fire on the germination potential of bitterbrush and yucca soil is due to increases in germination cue and nutrients. However, since the addition of cue and nutrients to the unburned soils under blackbrush and wolfberry shrubs did not elevate the germination potential of these soils to that found in burned soils, we conclude that these species alter the soil so as to inhibit germination and burning reverses this alteration. The presence of persistentN. attenuata populations in washes could not be attributed to particular chemical characteristics of these soils. Additions of cue dramatically increased germination potential of these soils, whereas the addition of nitrate did not. The concentrations of most mineral nutrients resembled those found in the unburned sites with the exception of Mn, Cu, and Zn, which were higher. However, these cations do not influence germination rates. Treatment of soil taken from these washes with cue resulted in significant increases in germination ofN. attenuata seeds in the natural seed bank compared to water-treated controls, demonstrating that the plants growing in washes also produce dormant seeds that require the smoke cue for germination.
47. A. Bano, N. Sharma, H. Dhaliwal, and V. Sharma, “A Systematic and Comprehensive Review on Withania Somnifera (L.) Dunal- An Indian Ginseng,” British Journal of Pharmaceutical Research, vol. 7, no. 2, pp. 63–75, Jan. 2015. doi: 10.9734/BJPR/2015/17102. Abstract
    Present review article reveals the importance of species Withania somnifera (L.) Dunal, distributed in India and other parts of the world, this extensive research information on this species is highly significant for future researchers worldwide. In this article cytomorphological, phytochemical and biological activities inputs have been extensively recorded and discussed. As a part of our investigation on cytomorphological and phytochemical aspects for important medicinal plants from India, the aim of this pioneer attempt is to provide precise, truthful and detailed information of W. somnifera (L.). As per our knowledge, there is not even a single, combined, constructive review report available about this species, evaluated by using cytomorphological, phytochemical and biological activities based aspects.
48. Banyan Botanicals, “Ashwagandha Root | Growing & Harvesting on Our Farm | Ayurvedic Herbs.” Nov-2019. https://www.youtube.com/watch?v=xu_3C5ImOg4.
49. S. S. Bargale, T. B. Tripathy, and H. K. Shashirekha, “Phyto Physicochemical Profile of Withania Somnifera Dunal (Solanaceae),” Journal of Drug Delivery and Therapeutics, vol. 9, no. 3-s, pp. 263–268, Jun. 2019. doi: 10.22270/jddt.v9i3-s.3008. Abstract
    Introduction- The present article deals with study of phytochemical analysis of Withania somnifera Dunal roots. Withania somnifera also known as Ashwagandha or winter cherry. Various preparations of Ashwagandha (WS) are available in the market used in the treatment of many clinical conditions in India. Objective- Evolution of Physico-chemical values and phytochemical analysis of Ashwagandha Churna.  Materials and Methods- The current investigation deals with extraction and detection or screening of active phytochemical compounds from different extracts of Withania somnifera root. Pharmacognostic studies, Physicochemical studies, Preliminary phytochemical studies was carried out. Result and conclusion - The result drown were 2% foreign mater was determined. Loss on drying 1.6%, total ash obtained was 9 %, acid insoluble ash was 1% and water soluble extractive was 12 % and Alcohol soluble extractive was 13 %. The phytochemical investigation revealed the presence of various phytochemical constituents such as alkaloids, flavonoids, carbohydrate, Steroids and Saponin Glycoside. Keywords: Ashwagandha, Withania Somnifera, Phytochemical.
50. D. Baricevic, A. Umek, S. Kreft, B. Maticic, and A. Zupancic, “Effect of Water Stress and Nitrogen Fertilization on the Content of Hyoscyamine and Scopolamine in the Roots of Deadly Nightshade (Atropa Belladonna),” Environmental and Experimental Botany, vol. 42, no. 1, pp. 17–24, Aug. 1999. doi: 10.1016/S0098-8472(99)00014-3. Abstract
    The study intended to elaborate the optimal environmental conditions of water supply and nitrogen fertilization for maximum content of hyoscyamine (% dw) and scopolamine (% dw). Plants grown from seeds of Slovene autochthonous population of deadly nightshade (Atropa belladonna), were treated with different water regimes (35–95% depletion of available soil water) together with enhanced nitrogen supply (0.37–1.60 g/pot N) in a greenhouse experiment. Dry plant extracts from 32-week old roots were analysed with capillary electrophoresis (CE) for the presence of tropane alkaloids (hyosciamyne, scopolamine). The results of the plant treatment responses showed that the maximal yield of tropane alkaloids (hyoscyamine: 54 mg/plant; scopolamine: 7 mg/plant) was achieved in plants grown under an optimal irrigation regime (35% depletion of available soil water) accompanied with total nitrogen supply of 0.37 g/pot. By contrast, the maximal content of alkaloids was achieved with 95% depletion of available soil water and a nitrogen supply of 1.60 g/pot.
51. D. A. Barnes, R. Barlow, P. S. Nigam, and R. K. Owusu-Apenten, “Antioxidant, Anticancer and Antibacterial Activity of Withania Somnifera Aqueous Root Extract,” Nov. 2015. doi: 10.9734/JABB/2016/22523. Abstract
    Aims: To evaluate total antioxidant capacity, anticancer activity and antibacterial effects Withania somnifera aqueous-root extracts. Study Design: In vitro study. Place of Study: School of Biomedical Sciences, Ulster University, UK. Methodology: Total antioxidant capacity (TAC) of whole powder and freeze dried W. somnifera aqueous-root extracts was determined using FRAP, DPPH, Folin and ABTS assays. Anticancer activity was accessed using MDA-MB-231 breast cells and Sulforhodamine B staining for cell viability. Antibacterial activity was by disk diffusion assay with penicillin, amoxicillin and streptomycin as positive controls. Results: The TAC for W. somnifera extract was 86, 47, 195,or 443 gallic acid equivalents per 100g dry basis (mgGAE/ 100 g) using FRAP, DPPH, Folin or ABTS assays, respectively. Corresponding TAC values for freeze dried W. somnifera aqueous-root extract were, 418, 553, 1898 or, 1770 (mgGAE/100 g). W. somnifera aqueous-root extract inhibited MDA-MB-231 cell proliferation in a dose-dependent manner with IC50 = 0.19 mg/ml (21 µM GAE). Nil antibacterial effects were detected for freeze dried W. somnifera extract (0-1 mg/ml) across six species of bacteria tested. Conclusion: Withania somnifera root water extract showed significant antioxidant and anticancer activity for MDA-MB-231 breast cancer cells but no antibacterial activity under the conditions of this study.
52. B. B. Basak, A. Saha, N. A. Gajbhiye, and P. Manivel, “Potential of Organic Nutrient Sources for Improving Yield and Bioactive Principle of Ashwagandha (Withania Somnifera) through Enhanced Soil Fertility and Biological Functions,” Communications in Soil Science and Plant Analysis, vol. 51, no. 6, pp. 779–793, Mar. 2020. doi: 10.1080/00103624.2020.1729368. Abstract
    Ashwagandha (Withania somnifera L. Dunal) is an industrially important medicinal herb and root and is a rich source of bioactive withanolides. To investigate the impacts of different organic nutrient management practices on ashwagandha yield and quality, a field experiment was conducted for 2 consecutive years. The experiment was carried out in randomized bock designed (RBD) with treatments comprised of different organic manures (farmyard manure, vermicompost, and castor cake) and microbial consortium along with recommended doses of fertilizer and the control. The root yield parameters (root girth, length, and fresh and dry root weight) of ashwagandha were influenced significantly by the treatments containing organic manures and microbial consortium alone or in combination. The highest fresh (1505 kg ha−1) and dry (767 kg ha−1) root yields as well as total withanolide contents (0.947 mg g−1) were recorded under the treatment receiving castor cake + microbial consortium (T7) followed by application of vermicompost + microbial consortium (T8). Significant improvements in soil fertility status (mineral N, available P and S) and soil biochemical parameters (microbial biomass and enzymes) were observed under the treatments containing castor cake and vermicompost. A positive correlation coefficient (< 0.01) was found between dry root yield, withanolide content and soil properties (r = 0.34–0.64). Thus, the results indicate that castor cake and vermicompost could be promising nutrient source for production of quality medicinal herb by improving soil properties.
53. H. S. Bashir, A. M. Mohammed, A. S. Magsoud, and A. M. Shaoub, “Isolation of Three Flavonoids from Withania Somnifera Leaves (Solanaceae) and Their Antimicrobial Activities,” Journal of Frest Products and Industries, vol. 2, no. 5, pp. 39–45, 2013. https://www.academia.edu/7216518/Isolation_of_Three_Flavonoids_from_Withania_Somnifera_Leaves_Solanaceae_and_their_Antimicrobial_Activities. Abstract
    Isolation of Three Flavonoids from Withania Somnifera Leaves (Solanaceae) and their Antimicrobial Activities
54. C. Baskaran and S. Velu., “Phytochemical Analysis and In-Vitro Antimicrobial Activity of Withania Somnifera (Ashwagandha),” Journal of Natural Products and Plant Resources, vol. 2, no. 6, pp. 711–716, 2012. https://www.scholarsresearchlibrary.com/abstract/phytochemical-analysis-and-invitro-antimicrobial-activity-ofrnwithania-somnifera-ashwagandha-6252.html. Abstract
    To investigate the antimicrobial activity and phytochemical screening Ethanol, methanol, Ethyl acetate, acetone, chloroform, Petroleum ether, hexane, hot water, and extracts of Withania somnifera. The aim of the present study was to evaluate the qualitative analysis of phytochemicals and antimicrobial activity of various solvent extracts of Withania somnifera. The antimicrobial activities of different solvent extracts of Withania somnifera were tested against the bacterial strains and fungus by observing the zone of inhibition. The Gram-positive bacteria used in the test were Staphylococcus aureus, Bacillus cereus and Micrococcus luteus, and the Gram-negative bacteria were Escherichia coli, Pseudomonas aeruginosa and Klebsiellapneumoniae, fungus like Aspergillusniger, Aspergillusflavus, Candida albicans, Candida tropicalis, Cryptococcus neoformans, and Candida kefyr. It was observed that ethanol; methanol, ethyl acetate, aceton, chloroform, petroleum ether, hexane and aquas extract showed activity against bacteria and fungus. The Ethanol extract of Withania somnifera showed more activity against Staphylococcus aureus, zone of diameter 20.10±0.17 mm and Methanol extract of Withania somnifera showed more activity against Candida albicans, zone of diameter 14.20±0.40 mm compared to other solvent extracts. In this study Ethanol extract in bacteria and methanol extract in fungus showed a varying degree of inhibition to the growth of tested organism, than Ethyl acetate, chloroform, aceton, Petroleum ether, hexane and hot water extracts.
55. S. Bastani et al., “Nano Iron (Fe) Complex Is an Effective Source of Fe for Tobacco Plants Grown under Low Fe Supply,” Journal of soil science and plant nutrition, vol. 18, no. 2, pp. 524–541, Jun. 2018. doi: 10.4067/S0718-95162018005001602. Abstract
    Abstract:In order to compare the uptake, utilization and distribution of iron (Fe) as nano and bulk Fe complex (Fe(III)-EDTA), an experiment was conducted using hydroponically-grown Fe-deficient tobacco (Nicotiana rustica L.) plants. Plants were treated with Fe either through roots (root application, RA) or leaves (foliar application, FA). Leaf chlorophyll concentration and plants biomass responded to the Fe re-supply; this response was quicker for the nano Fe complex than the bulk Fe complex, in particular, in the RA plants. Plants re-supplied with the nano Fe complex had lower Fe content but higher Fe use efficiency than plants re-supplied with the bulk Fe complex. Analysis of different plant fractions at two subsequent weeks revealed that the nano Fe complex had higher mobility than the bulk Fe complex, both in the xylem and the phloem. Comparison of RA and FA revealed that the re-supply of Fe through roots was more efficient in retrieving the whole plant growth than the foliar Fe spray. RA, in contrast to FA plants, profited from an enhanced Fe uptake capacity induced under Fe starvation in the roots. Alternatively, FA was effective in extending the green leaf area duration. Our data suggested that nano Fe complex is advantageous as both leaf spray and a long-term feeding of plant through roots. Keywords: Utilization; efficiency; partitioning; retranslocation; leaf age
56. A. Bennici, “Stimulation of Germination, Callus Growth and Shoot Regeneration of Nicotiana Tabacum L. by Pulsing Electromagnetic Fields (PEMF),” Stimulation of germination, callus growth and shoot regeneration of Nicotiana tabacum L. by Pulsing Electromagnetic Fields (PEMF), pp. 1000–1004, 2000. doi: 10.1400/14034. Abstract
    Purchase online the PDF of Stimulation of germination, callus growth and shoot regeneration of Nicotiana tabacum L. by Pulsing Electromagnetic Fields (PEMF), Bennici, A.,Bovelli, R. - Firenze University Press - Article
57. R. Bessalle and D. Lavie, “Semi-Quantitative Reversed-Phase High-Performance Liquid Chromatographic Analysis of the Ecotypes of Withania Somnifera Chemotype III,” Journal of Chromatography A, vol. 389, pp. 195–210, Jan. 1987. doi: 10.1016/S0021-9673(01)94423-1. Abstract
    Eleven withanolides isolated from Withania somniferal. (Dun.) were analyzed on a C18 reversed-phase column. Good separation factors were obtained, especially when analysing the major component of the plant. The semi-quantitative determination of the main withanolides of chemotype III collected from four different locations (Yavne, Gedera, Farm-1 and Farm-2) was carried out in order to confirm the presence of ecotypes. The standards were obtained by isolation and purification of the withanolides from the ecotype identified as Farm-2 and our own collection. As a result of the semi-quantitative analysis, three ecotypes were found. They were identified according to the differences in concentration of their major component, and were related to the various locations.
58. R. Bessalle, D. Lavie, and F. Frolow, “Withanolide Y, a Withanolide from a Hybrid of Withania Somnifera,” Phytochemistry, vol. 26, no. 6, pp. 1797–1800, Jan. 1987. doi: 10.1016/S0031-9422(00)82291-8. Abstract
    The structure of a withanolide isolated from the hybrid of Withania somnifera chemotype III (Israel) by Indian I (Delhi) has been elucidated as (20R,22R)-5α,6α-epoxy-7α-17α,20-trihydroxy-1-oxo-witha-2,24-dienolide by X-ray single crystal analysis. This compound is the first example of a 5α,6α-epoxy-7α-hydroxy system among the withanolides, and thus provides an additional step in the biosynthetic pathway. Its reaction with trichloroacetyl isocyanate has been followed by 1H NMR and compared with withanolide T.
59. R. Bessalle and D. Lavie, “Withanolide C, A Chlorinated Withanolide from Withania Somnifera,” Phytochemistry, vol. 31, no. 10, pp. 3648–3651, Oct. 1992. doi: 10.1016/0031-9422(92)83749-O. Abstract
    The structure of a new chlorinated withanolide isolated from Withania somnifera chemotype III has been elucidated as 5α-chloro,6β,14α,17β,2OαF-tetrahydroxy-l-oxo-22R-witha-2,24-dienolide by 1H and 13CNMR and chemical ionization mass spectroscopy methods.
60. T. Bharathi, S. Gnanamurthy, D. Dhanavel, and M. Ariraman, “Induced Physical Mutagenesis and Its Effect in Cytological Behavior of Ashwagandha ( Withania Somnifera (L.) Dunal),” International Letters of Natural Sciences, vol. 17, pp. 152–161, Jun. 2014. doi: 10.18052/www.scipress.com/ILNS.17.152. Abstract
    The mitotic effect of physical mutagen gamma rays was observed in the root tip cells of Ashwagandha. The Chromosome analysis has been showed as an important tool for establish variability of the plant seed by use of physical mutagen gamma rays. The gamma rays have of low wavelength and high penetrable power. The plant has tremendous medicinal values and it is known from ancient times. The dry and well matured seeds of ashwagandha were irradiated with different doses of gamma rays viz., 5, 10, 15, 20, 25, 30, 35, 40, 45 and 50 KR respectively. The chromosome number of control plant is 2n = 48. The gamma rays affect the normal cytological behavior of ashwagandha species. The chromosomal aberrations increase with increase in the doses of gamma rays to optimum level of 30KR, because it causes changes in the chromosome structure, cellular structure and metabolism of plants. The chromosome aberration like, Sticky metaphase, Precocious moment chromosome, Fragments, Anaphasic bridge, Anaphasic laggard, Telophasic laggard. The present investigation was carried out to study the cytogenetic analysis of the species Withania somnifera. The chromosomal aberration increases with increase in the doses to optimum level (50 KR) of physical mutagen gamma rays.
61. S. K. Bharti, A. Bhatia, S. K. Tewari, O. P. Sidhu, and R. Roy, “Application of HR-MAS NMR Spectroscopy for Studying Chemotype Variations of Withania Somnifera (L.) Dunal,” Magnetic Resonance in Chemistry, vol. 49, no. 10, pp. 659–667, 2011. doi: 10.1002/mrc.2817. Abstract
    Withania somnifera (L.) Dunal (Solanaceae), commonly known as Ashwagandha, is one of the most valued Indian medicinal plants with a number of pharmaceutical and nutraceutical applications. Metabolic profiling has been performed by HR-MAS NMR spectroscopy on fresh leaf and root tissue specimens from four chemotypes of W. somnifera. The HR-MAS NMR spectroscopy of lyophilized defatted leaf tissue specimens clearly distinguishes resonances of medicinally important secondary metabolites (withaferin A and withanone) and its distinctive quantitative variability among the chemotypes. A total of 41 metabolites were identified from both the leaf and root tissues of the chemotypes. The presence of methanol in leaf and root tissues of W. somnifera was detected by HR-MAS NMR spectroscopy. Multivariate principal component analysis (PCA) on HR-MAS 1H NMR spectra of leaves revealed clear variations in primary metabolites among the chemotypes. The results of the present study demonstrated an efficient method, which can be utilized for metabolite profiling of primary and secondary metabolites in medicinally important plants. Copyright © 2011 John Wiley & Sons, Ltd.
62. S. Bhasin, M. Singh, and D. Singh, “Review on Bioactive Metabolites of Withania Somnifera. (L.) Dunal and Its Pharmacological Significance,” Journal of Pharmacognosy and Phytochemistry, vol. 8, no. 3, pp. 3906–3909, 2019. https://www.researchgate.net/publication/344189197_Review_on_bioactive_metabolites_of_Withania_somnifera_L_Dunal_and_its_pharmacological_significance. Abstract
    Naturally active compounds from the medicinal plants are used by tribal people for many disorders. Ayurvedic medicines are most commonly used. Withania somnifera (L.) is an important medicinal plant which is widely used for the treatment of various disorders like bronchitis, ulcer, inflammatory diseases, and various stomach problems. It is a challenge to extract the bioactive plant constituent from the different parts of a plant. W. somnifera and its constituent have a remarkable biological activity which is appropriate to develop medicine and can be chemically modified and biotransform to make a more potent medicine. Numbers of in vitro and in vivo experiments have been demonstrated to find the ability of W. somnifera. This plant is a good source of bioactive constituents like withanolides and withaferin which are used for the treatment of cancer. The major phytoconstituents of this species are steroidal lactones.
63. A. Bhatia, S. K. Bharti, S. K. Tewari, O. P. Sidhu, and R. Roy, “Metabolic Profiling for Studying Chemotype Variations in Withania Somnifera (L.) Dunal Fruits Using GC–MS and NMR Spectroscopy,” Phytochemistry, vol. 93, pp. 105–115, Sep. 2013. doi: 10.1016/j.phytochem.2013.03.013. Abstract
    Withania somnifera (L.) Dunal (Solanaceae), commonly known as Ashwagandha, is one of the most valued Indian medicinal plant with several pharmaceutical and nutraceutical applications. Metabolic profiling was performed by GC–MS and NMR spectroscopy on the fruits obtained from four chemotypes of W. somnifera. A combination of 1H NMR spectroscopy and GC–MS identified 82 chemically diverse metabolites consisting of organic acids, fatty acids, aliphatic and aromatic amino acids, polyols, sugars, sterols, tocopherols, phenolic acids and withanamides in the fruits of W. somnifera. The range of metabolites identified by GC–MS and NMR of W. somnifera fruits showed various known and unknown metabolites. The primary and secondary metabolites observed in this study represent MVA, DOXP, shikimic acid and phenylpropanoid biosynthetic metabolic pathways. Squalene and tocopherol have been rated as the most potent naturally occurring compounds with antioxidant properties. These compounds have been identified by us for the first time in the fruits of W. somnifera. Multivariate principal component analysis (PCA) on GC–MS and NMR data revealed clear distinctions in the primary and secondary metabolites among the chemotypes. The variation in the metabolite concentration among different chemotypes of the fruits of W. somnifera suggest that specific chemovars can be used to obtain substantial amounts of bioactive ingredients for use as potential pharmacological and nutraceuticals agents.
64. R. S. Bhosale and A. D. More, “Response of Withania Somnifera, (L.) Dunal. to Soils from Different Locations in Satara District w.r.t Germination and Vegetative Growth,” International Journal of Life Sciences, vol. 1, no. 1, pp. 76–78, 2013. Abstract
    Withania somnifera, Dunal. belongs to family Solanaceae. It is also known as Ashwagandha or Indian Genseng. Withanolides are constituents of roots of Ashwagandha. Withanolide is the major constituent with appreciable quantity of Withanone and traces of Withaferin-A. In Ayurvedic literature, is a “Rasayana” or rejuvenating drug and hence also traditionally known as Avarada which suggests the application of this plant for enhancing longevity. The seeds of Withania somnifera, Dunal. var somnifera were selected and used for following investigations. Different locations from Junnar taluka showed different response to germination and vegetative growth. From present investigation it can be concluded that soil with alkaline properties was more efficient for germination and growth of Withania somnifera, Dunal. than acidic soil.
65. P. von Bieberstein, Y.-ming Xu, A. A. L. Gunatilaka, and R. Gruener, “Biomass Production and Withaferin A Synthesis by Withania Somnifera Grown in Aeroponics and Hydroponics,” HortScience, vol. 49, no. 12, pp. 1506–1509, Dec. 2014. doi: 10.21273/HORTSCI.49.12.1506. Abstract
    The medicinal herb Withania somnifera (L.) Dunal (Solanaceae) was grown in two soilless systems to determine optimal conditions for production of biomass and withaferin A, the major secondary metabolite responsible for its claimed medicinal properties. Withaferin A content was analyzed using high-performance liquid chromatography (HPLC). The results show that there was no statistically significant difference (P > 0.05; t test) in biomass production between the plants grown aeroponically and hydroponically. Aeroponically grown plants produced an average of 49.8 g dried aerial plant material (DW) (sd 20.7) per plant, whereas hydroponically grown plants produced an average of 57.6 g DW (sd 16.0). In contrast, withaferin A content was statistically higher in plants grown hydroponically. These plants contained an average of 7.8 mg·g−1 DW (sd 0.3), whereas the aeroponically grown plants contained an average of 5.9 mg·g−1 DW (sd 0.6). These results demonstrate that hydroponic techniques are optimal in reproducibly and efficiently generating withaferin A. These findings may be of importance to the natural products industry in seeking to maximize production of biologically active compounds from medicinal plants.
66. Bilal Ahmad Mir, “Reproductive Behaviour and Breeding System of Wild and Cultivated Types of Withania Somnifera (L.) Dunal,” Journal of Medicinal Plants Research, vol. 6, no. 5, Feb. 2012. doi: 10.5897/JMPR11.1303. Abstract
    The Indian germplasm of Withania somnifera (L.) Dunal shows remarkable genetic variability both in the cultivated and the wild populations. The utilization of this variability through conventional breeding requires a clear understanding of its reproductive biology and breeding system. It is an amphimictic species practicing open pollination. However, a proximal placement of the stigma and the anther and a synchrony between the receptivity and dehiscence of anthers strongly predispose the species to self pollination and selfing. However open pollination results in equally high percentage of fruit and seed set as on controlled selfing indicating that the species shows facultative autogamy. This raises the probability of genetic improvement through hybridization. The somatic complements revealed a diploid number of 2n=48 in all the accessions tested. The absence of karyomorphological differences indicated that numerical and structural changes do not have a role in controlling the genetic variability of the species. Experimental crosses between the cultivated and the wild accessions produced viable seeds. A significantly higher fruit set and seed germ inability in crosses involving the cultivated types as the seed parent point to the existence of maternal effect.
67. R. Bimal, “In Vitro Induction of Germination in Ovules of Nicotiana Plumbaginifolia Viv.,” Proceedings of the international congress of plant physiology, New Dehli, India, 15-20 February 1988. Volume 2., pp. 1366–1368, 1990. https://www.cabdirect.org/cabdirect/abstract/19920316059. Abstract
    The dependence of fertilized ovule growth on maternal tissues (placentas and ovary wall) was studied in vitro in N. plumbaginifolia. A precise developmental sequence of ovules, placentas and ovary wall was essential for the uninterrupted growth of attached ovules on MS medium in vitro. The ovules attached to maternal tissues matured into seeds and germinated on MS basal medium. However, addition...
68. A. D. Blonstein, P. Stirnberg, and P. J. King, “Mutants of Nicotiana Plumbaginifolia with Specific Resistance to Auxin,” Molecular and General Genetics MGG, vol. 228, no. 3, pp. 361–371, Sep. 1991. doi: 10.1007/BF00260628. Abstract
    We have isolated nine independent auxin-resistant mutants of Nicotiana plumbaginifolia by culturing M2 seedlings in the presence of indole-3-acetic acid ethyl ester or 1-naphthaleneacetic acid at concentrations which significantly inhibit hypocotyl elongation of the wild type. The mutations were induced by treating seed with ethyl methanesulphonate and were found in the course of screening 10 000 individual M2 families. Auxin resistance was in all cases the result of a mutation at a single, nuclear locus. The dominance relationships of two of the mutants could be defined as recessive or dominant; all other mutants showed partial dominance. In contrast to previously described mutants of Arabidopsis and N. plumbaginifolia, all of the present mutants were specifically resistant to auxin; the mutants were cross-resistant to several auxins, but showed no increased resistance to cytokinin, abscisic acid, ethylene or 1-amino-cyclopropane-1-carboxylic acid. The importance of the choice of the selection criterion for the isolation of specific resistance traits is discussed.
69. D. Bocciarelli, “Medium Optimization for the Rhizosecretion of Three Monoclonal Antibodies from Nicotiana Tabacum Hydroponic Culture,” Other, Université de Lorraine, 2014. https://hal.univ-lorraine.fr/hal-01835626. Abstract
    Plants are being developed as an inexpensive alternative platform for the production of recombinant pharmaceuticals, including monoclonal antibodies (mAbs). Transgenic plants were previously demonstrated to secrete the recombinant proteins from their roots in a process called rhizosecretion. Thus hydroponic cultivation of transgenic plants that allows harvesting of recombinant proteins from the hydroponic medium has been investigated for some years, but has not yet provided sufficient yields of target proteins for commercial viability. Scaling up the current in vitro production system could consist of the elaboration of an appropriate media for Nutrient Film Technique (NFT) ex vitro system. Medium supplemented with gelatin has previously shown 20 times-improvement of rhizosecreted antibody yields but is incompatible with NFT. In this project we have assessed loss in yields in the absence of gelatin of three monoclonal antibodies: M12 (anti-vitronectin), VRC01 (anti- HIV) and ?E559 (anti-rabies). Consistently, a 50% yield decrease occurred without gelatin, for each antibody. The NFT system has so far been recalcitrant to rhizosecretion with no detectable yields. Nitrate enriched hydroponic medium has previously been found to increase antibody secretion, by a mechanism currently thought to be osmolarity-related. A similar enrichment was applied to NFT medium in vitro in this project, and for the first time M12 antibody secretion could be detected and quantified. These results are encouraging for the scaling up of rhizosecretion to the NFT system as a recombinant protein production platform. Presently, plant yields remain dramatically lower than mammalian-cell production and plant molecular farming has not been embraced by pharmaceutical companies. However, such plant manufacturing systems could be of particular interest for the development of small scale, low-technology recombinant therapeutics production systems, highly needed to fight neglected diseases (e.g. rabies, tuberculosis) occurring in developing countries.
70. J. Bolleddula, W. Fitch, S. K. Vareed, and M. G. Nair, “Identification of Metabolites in Withania Sominfera Fruits by Liquid Chromatography and High-Resolution Mass Spectrometry,” Rapid Communications in Mass Spectrometry, vol. 26, no. 11, pp. 1277–1290, 2012. doi: 10.1002/rcm.6221. Abstract
    RATIONALE Withania somnifera is a rich source of biologically active secondary metabolites. Our earlier investigations on the fruits of this plant have yielded novel compounds, withanamides, with potent antioxidant activity and protective effect on β-amyloid-induced cytotoxicity in vitro. However, several minor compounds present in the fruits have not been characterized previously which may contribute to the observed biological activities. METHODS Liquid chromatography (LC) coupled with high-resolution mass spectrometry (HRMS) with data-dependent and targeted MS/MS experiments were conducted to elucidate the structure of observed metabolites. RESULTS A total of 62 metabolites identified included 32 withanamides, 22 withanolides, 3 steroidal saponins, 2 lignanamides, feruloyl tyramine, methoxy feruloyl tyramine and a diglucoside of hydroxyl palmitic acid. The structures of these compounds were proposed based on accurate masses of the molecular and fragment ions. Several of these new compounds identified from the profile were derived from withanamides with variations in aliphatic and/or glycosyl moieties. In addition, six new withanolides, a new hydroxy fatty acid diglucoside and several known compounds in the extract were identified. CONCLUSIONS The current study revealed the presence of several new and known compounds in Withania somnifera fruits. High-resolution MS and MS/MS experiments provide an extremely effective approach to derive the structures of plant secondary metabolites including isomeric compounds. Copyright © 2012 John Wiley & Sons, Ltd.
71. D. A. Bonilla, Y. Moreno, C. Gho, J. L. Petro, A. Odriozola-Martínez, and R. B. Kreider, “Effects of Ashwagandha (Withania Somnifera) on Physical Performance: Systematic Review and Bayesian Meta-Analysis,” Journal of Functional Morphology and Kinesiology, vol. 6, no. 1, p. 20, Mar. 2021. doi: 10.3390/jfmk6010020. Abstract
    Ashwagandha (Withania somnifera) is considered a potent adaptogen and anti-stress agent that could have some potential to improve physical performance. This preferred reporting items for systematic reviews and meta-analyses (PRISMA)-based comprehensive systematic review and Bayesian meta-analysis aimed to evaluate clinical trials up to 2020 from PubMed, ScienceDirect, and Google Scholar databases regarding the effect of Ashwagandha supplementation on physical performance in healthy individuals. Besides implementing estimation statistics analysis, we developed Bayesian hierarchical models for a pre-specified subgroup meta-analysis on strength/power, cardiorespiratory fitness and fatigue/recovery variables. A total of 13 studies met the requirements of this systematic review, although only 12 were included in the quantitative analysis. A low-to-moderate overall risk of bias of the trials included in this study was detected. All Bayesian hierarchical models converged to a target distribution (Ȓ = 1) for both meta-analytic effect size (μ) and between-study standard deviation (τ). The meta-analytic approaches of the included studies revealed that Ashwagandha supplementation was more efficacious than placebo for improving variables related to physical performance in healthy men and female. In fact, the Bayesian models showed that future interventions might be at least in some way beneficial on the analyzed outcomes considering the 95% credible intervals for the meta-analytic effect size. Several practical applications and future directions are discussed, although more comparable studies are needed in exercise training, and athletic populations are needed to derive a more stable estimate of the true underlying effect.
72. H. D. Boswell, A. B. Watson, N. J. Walton, and D. J. Robins, “Formation of N′-Ethyl-S-Nornicotine by Transformed Root Cultures of Nicotiana Rustica,” Phytochemistry, vol. 34, no. 1, pp. 153–155, Aug. 1993. doi: 10.1016/S0031-9422(00)90797-0. Abstract
    N-Ethylputrescine dihydrochloride has been synthesized by an improved procedure and it is converted by transformed root cultures of Nicotiana rustica into the nicotine analogue, N′-ethyl-S-nornicotine, preferentially in the optically active S-form, with an efficiency similar to that of the corresponding natural process.
73. H. D. Boswell et al., “Metabolism of N-Alkyldiamines and N-Alkylnortropinones by Transformed Root Cultures of Nicotiana and Brugmansia,” Phytochemistry, vol. 52, no. 5, pp. 855–869, Nov. 1999. doi: 10.1016/S0031-9422(99)00292-7. Abstract
    A range of analogues of N-methylputrescine and tropinone were fed to transformed root cultures of Nicotiana rustica and/or a Brugmansia candida×aurea hybrid. These cultures were made by the transformation of the relevant plant species with Agrobacterium rhizogenes. A number of the metabolites, notably those showing a relatively modest alteration in the N-alkyl substituent, were metabolized in vivo to form homologues of the normal alkaloids biosynthesized by these roots. These products were identified by GC/MS and comparison with some synthetic reference materials. Analogues with major alterations in the size of the N-alkyl substituent were not metabolized at all. In the N. rustica cultures, the analogues fed at 1 mM significantly affected the profile of normal alkaloids, with up to a 4-fold diminution in nicotine being found in the presence of N-n-propylputrescine. The ratio between alkaloids of the pyrrolidine series and the piperideine series was also affected. In contrast, the presence of the analogues in the B. candida×aurea hybrid culture at 1 mM did not inhibit or substantially interfere with the accumulation of the normal spectrum of alkaloids. The potential for using these cultures to make complex novel products from simple precursors is discussed.
74. H. J. Boughey, “Competitive Ability and Environmental Sensitivity in Nicotiana Rustica.,” PhD thesis, University of Birmingham, 1978. https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.479642. Abstract
    In the first part of this thesis the effects of natural selection on two contrasting synthetic populations, of known initial genetical composition, after successive generations of mating in isolated, competitive conditions, are investigated for a number of quantitatively inherited characters. Predictions of the magnitude and direction of the changes expected are made from a knowledge of the genetical architecture of the traits, from the relative fitnesses of the genotypes of the initial population and from estimates of the breedirgsystem operating in successive generations of the population. The accuracy of these predictions, and the validity of, the underlying genetical theory and assumptions, are tested by monitoring the observed changes occurring in the two experimental, synthetic populations. Sensitivity to macro-environmental variation between seasons may be a major determinant of the competitive ability of the competing genotypes, so in the second part the measurement and prediction of sensitivity and its determinants arestudied. Joint selection is performed for both extremes of mean performance and sensitivity to macro-environmental differences in several seasons, using family selection, and in a number of environments within the same, season, using lines derived by single seed descent. Indirect selection, as a practical alternative to direct selection, through the correlation between sensitivity and pattern of growth, as summarised by the quadratic component of the growth curve and the ratio of height in the middle of the season to final height, is investigated.
75. D. Bouthour, A. Hajjaji-Nasraoui, L. Saafi, H. Gouia, and C. Chaffei-Haouari, “Effects of NaCl on Growth and Activity of Enzymes Involved in Carbon Metabolism in Leaves of Tobacco (Nicotiana Rustica),” African Journal of Biotechnology, vol. 11, no. 63, pp. 12619–12629, 2012. doi: 10.4314/ajb.v11i63. Abstract
    The adverse effects of salt should not be the same in tobacco plants exposed to a permanent and transient high concentration of NaCl in its environment. Experiments were conducted in order to verify the hypothesis of reversibility of NaCl effects. The study of this reversibility is checked by monitoring a number of parameters in pre-stressed plants and then, replaced in normal conditions. Plants previously grown for 30 days on basic medium were treated for 7 days with 200 mM NaCl and then placed back on the basic culture without NaCl for 10 days. The results show that NaCl suppression leads to a resumption of growth with a decrease in the concentration of sodium (Na+) and chloride ions (Cl-). Hence, potassium content (K+) increases gradually in the leaves to reach the level obtained with unstressed plants. At the same time, there is a stimulation of the activities of phosphoenolpyruvate carboxylase (PEPC), malate dehydrogenase isoenzymes (NAD, NADP, NADH and NADPH-MDH) and isocitrate dehydrogenase (ICDH) after NaCl had been removed. Along with the boosting of the activity of these enzymes involved in the process of carbon assimilation, there is a gradual decrease in soluble sugars content, suggesting a resumption of the normal activity of photosynthetic assimilation process. All these results verify our hypothesis and can be explained by the ability of the plant to dilute the effects of Na+ and Cl- during the recovering period. An important result of this study is that a transient salinity is not necessarily followed by a significant depreciation in product yield or quality.Keywords: Tobacco, NaCl, reversibility, phosphoenolpyruvate carboxylase (PEPC), malate dehydrogenase (MDH), isocitrate dehydrogenase (ICDH)
76. E. L. Breese, “Selection for Differing Degrees of Out-Breeding in Nicotiana Rustica,” Annals of Botany, vol. 23, no. 2, pp. 331–344, Apr. 1959. doi: 10.1093/oxfordjournals.aob.a083659. Abstract
    Initial observations on Nicotiana rustica suggested that the percentage of seed set by outcrossing might vary considerably in a segregating population and that this might be due to variation in the position of the stigma relative to the anthers in the corolla-tube. This, measured as the difference in level between the stigma and the ring of anthers, is termed heterostathmy and the average expression of this character could be significantly altered by a short programme of selection. These structural modifications were accompanied by developmental modifications affecting the timing of anther dehiscence relative to the opening of the flower. Tests indicated that these changes significantly affected the rate of outbreeding.
77. W. R. Brewer and L. D. Hiner, “Cultivation Studies of the Solanaceous Drugs. II. The Effect of Nutritional and Soil Reaction Fertilizers on the Production Yields and Total Alkaloidal Content of Atropa Belladonna and Hyoscyamus Niger,” Journal of the American Pharmaceutical Association, vol. 39, no. 10, pp. 586–591, 1950. doi: 10.1002/jps.3030391015. Abstract
    In order to clarify some of the contradiction and confusion existing in the literature concerning the value of adjusting pH, and nitrogen, phosphorus, and potassium levels in solanaceous plants and their alkaloid production, a study was begun with belladonna and hyoscyamus plants growing in controlled variations of these elements and their concentrations. The results of these initial studies were subjected to statistical interpretation to determine their significance.
78. M. P. Bridgen and R. E. Veilleux, “A Comparison of in Vitro Flowers to in Vivo Flowers of Haploid and diploidNicotiana Tabacum L.,” Plant Cell, Tissue and Organ Culture, vol. 13, no. 1, pp. 3–13, Jan. 1988. doi: 10.1007/BF00043042. Abstract
    Thin cell layers (TCLs) were cultured from inflorescences of diploid (2n=4x=48) and haploid (2n=2x=24)Nicotiana tabacum L. "Samsun" and the subsequent flowers formed in vitro were then compared to in vivo flowers. Plants derived from TCLs possessed flowers that were typical of their seed or androgenetically-derived counterparts, whereas de novo flowers from TCLs were abnormal when compared to their counterparts. The TCLs of haploid plants produced more flower buds than diploid TCLs, and did so in a shorter period of time. In vitro flowers and anthers at both ploidy levels were considerably smaller than the in vivo flowers; in vitro flowers also had variable numbers of anthers and pistils. The embryogenic capacity of anthers taken from in vivo diploid flowers was 5 times greater than that of in vitro diploid or haploid anthers. In vivo haploid anthers produced no embryoids, whereas in vitro haploid anthers did produce embryoids. Observations of mitotic cells in root tips of plants derived from anther cultures of in vitro haploid flowers revealed a mixoploid nature. Diploid meiosis was regular and haploid meiosis was irregular regardless of the origin (in vitro or in vivo) of the flowers.
79. M. P. Bridgen, “Studies of in Vitro Flowering and de Novo Flowers of Nicotiana Tabacum,” 1984. https://vtechworks.lib.vt.edu/handle/10919/76061. Abstract
    The objectives of this research were to examine factors influencing de novo flowering of Nicotiana on 2-3 x 10mm explants consisting of epidermal and 3-6 layers of subjacent cells (thin cell layers, TCLs) and to compare de novo to in vivo flowers. TCLs from short-day and long-day tobacco plants were compared with TCLs from day-neutral species to examine in vitro floral photoinduction and graft transmissibility of floral promoters and inhibitors. TCLs from photoperiodic species of tobacco did not form flowers de novo , whereas TCLs from day-neutral plants did flower. When TCLs were removed from photoperiodic plants and grafted in vitro to TCLs from day-neutral plants, there was no indication that a floral-promoter or inhibitor was transported through the non-vascular graft union. In vitro photoinduction of TCLs removed from photoperiodic plants was not possible under conditions conducive to in vitro flowering of TCLs from day-neutral species. TCLs taken from intraspecific F₁ and F₂ hybrids between short-day and day-neutral cultivars of N. tabacum were examined to assess the importance of genotype and photoperiod to de novo flowering. Flowering of the F₂ population occurred over a 9 week period under naturally decreasing photoperiod. Photoperiodic response and in vitro flowering were correlated in the F₂ population with fewer flowers produced per TCL with increasing short-day reaction. F₂ segregates whose TCLs did not yield de novo flowers were found among both day-neutral and short-day phenotypes. When de nova flowers were compared to in vivo flowers of diploid (2n=4x=48) N. tabacum ’Samsun’ and haploid (2n=2x=24) plants derived from ’Samsun’ anther culture, major morphological differences were found. Flower and anther sizes were reduced in de novo flowers and the numbers of anthers and pistils produced per flower were variable. TCLs from haploid plants produced more flowers in a shorter period of time than TCLs from diploid plants. Anthers cultured from de novo haploid plants were embryogenetic resulting in mixoploid plants; anthers from in vivo haploid flowers were not embryogenetic. Anthers from in vivo diploid plants were five times more embryogenetic than anthers from either de novo haploid or diploid flowers. Meiotic analysis revealed similar abnormalities from both in vivo and de novo microsporogenesis of haploids.
80. J. L. Bronstein, T. Huxman, B. Horvath, M. Farabee, and G. Davidowitz, “Reproductive Biology of Datura Wrightii: The Benefits of a Herbivorous Pollinator,” Annals of Botany, vol. 103, no. 9, pp. 1435–1443, Jun. 2009. doi: 10.1093/aob/mcp053. Abstract
    A deeper understanding of mutualism can be reached by studying systems with measurable costs and benefits. Most studies of this type focus on an unusual class of obligate, species-specific pollination mutualisms. The interaction between Datura wrightii (Solanaceae) and the hawkmoth Manduca sexta offers similar advantages but greater generality. Adult moths both nectar at and deposit eggs on the same plant; larvae are herbivorous. The antagonistic component of this interaction has been well studied. Here the role of M. sexta as a pollinator of D. wrightii, particularly in the context of this moth’s frequent nectaring visits to the bat-pollinated plant Agave palmeri, is documented.Hand-pollinations were used to determine breeding system and the reproductive consequences of mixed loads of A. palmeri and D. wrightii pollen. Plants and moths were caged overnight to assess whether nectaring visits led to fruit and seed set. Finally, pollen deposited on field-collected stigmas was identified, with a particular focus on documenting the presence of D. wrightii and A. palmeri grains.Datura wrightii is highly self-compatible, and a visit that deposits either outcross or self pollen almost doubles fruit and seed set compared with unvisited flowers. Manduca sexta transferred enough pollen to produce fruit and seed sets comparable to hand-pollination treatments. Agave palmeri did not interfere with D. wrightii success: in the field, stigmas received almost pure D. wrightii pollen, and hand-addition of large quantities of A. palmeri pollen had no measurable effect on fruit and seed set.The floral visitation component of the D. wrightii–M. sexta interaction is indeed mutualistic. This finding is essential background to future development of this interaction as a model system for studying mutualism’s costs and benefits. It is already proving valuable for dissecting third-species effects on the outcome of mutualism. Results indicate that M. sexta’s heavy visitation to A. palmeri has no negative effect on the benefits conferred to D. wrightii. However, it can be predicted to augment M. sexta populations to the point where the costs of the interaction begin to exceed its benefits.
81. S. A. Brown and R. U. Byerrum, “The Origin of the Methyl Carbon of Nicotine Formed by Nicotiana Rustica L. ¹,” Journal of the American Chemical Society, vol. 74, no. 6, pp. 1523–1526, Mar. 1952. doi: 10.1021/ja01126a052.
82. J. A. Burcev, “Breeding Nicotiana rustica varieties producing fewer suckers.,” Agrobiologija (Agrobiology), no. 3, pp. 345–49, 1960. https://www.cabdirect.org/cabdirect/abstract/19611605161. Abstract
    An account is given of breeding varieties which develop few or no secondary suckers together with a description of Malopasynkovaja 2 [Few-suckered 2], bred for this character at Simferopol’. The method consisted of crossing malformed hybrids of the type known as Omega, in this instance a hybrid of Indusskaja 15 [Indian 15] x Hmelovka, with an Omega plant of the variety Pozdniki. The development...
83. L. G. Burk and H. E. Heggestad, “The genusNicotiana: A Source of Resistance to Diseases of Cultivated Tobacco,” Economic Botany, vol. 20, no. 1, pp. 76–88, Jan. 1966. doi: 10.1007/BF02861929. Abstract
    Nicotiana tabacum L., cultivated tobacco, is one of 65 species in the genusNicotiana. Over the past 30 years, varying numbers of the species have been included in tests of resistance to tobacco diseases by research workers in tobacco-producing areas throughout the world. This report summarizes these investigations in tabular form and discusses methods whereby interspecific transfer of disease resistance may be accomplished.
84. H. R. Burton, R. A. Andersen, P. D. Fleming, and L. R. Walton, “Changes in Chemical Composition of Burley Tobacco during Sensecence and Curing. 2. Acylated Pyridine Alkaloids,” Journal of Agricultural and Food Chemistry, vol. 36, no. 3, pp. 579–584, May 1988. doi: 10.1021/jf00081a043.
85. H. R. Burton, G. H. Childs, R. A. Andersen, and P. D. Fleming, “Changes in Chemical Composition of Burley Tobacco during Senescence and Curing. 3. Tobacco-Specific Nitrosamines,” Journal of Agricultural and Food Chemistry, vol. 37, no. 2, pp. 426–430, Mar. 1989. doi: 10.1021/jf00086a034.
86. H. R. Burton and M. J. Kasperbauer, “Changes in Chemical Composition of Tobacco Lamina during Senescence and Curing. 1. Plastid Pigments,” ACS Publications, May 2002. doi: 10.1021/jf00065a028.
87. H. R. Burton, G. H. C. Jr, R. A. Andersen, and P. D. Fleming, “Changes in Chemical Composition of Burley Tobacco during Senescence and Curing. 3. Tobacco-Specific Nitrosamines,” ACS Publications. American Chemical Society, May-2002. doi: 10.1021/jf00086a034.
88. H. R. Burton, L. P. Bush, and M. V. Djordjevic, “Influence of Temperature and Humidity on the Accumulation of Tobacco-Specific Nitrosamines in Stored Burley Tobacco,” ACS Publications. American Chemical Society, May-2002. doi: 10.1021/jf00089a036.
89. R. W. Bussmann et al., “Nicotiana Rustica L. Nicotiana Tabacum L. Solanaceae,” in Ethnobotany of the Mountain Regions of Far Eastern Europe : Ural, Northern Caucasus, Turkey, and Iran, K. Batsatsashvili, Z. Kikvidze, and R. W. Bussmann, Eds. Cham: Springer International Publishing, 2020, pp. 623–629. doi: 10.1007/978-3-030-28940-9_93. Abstract
    Nicotiana rustica L.: Nicotiana pavonii Dunal
90. F. A. Bynov, “The effect of topping and removing the side shoots on the formation of nicotine in Nicotiana rustica.,” Doklady Akademii nauk SSSR, vol. 73, pp. 833–6, 1950. https://www.cabdirect.org/cabdirect/abstract/19510303735. Abstract
    In trials involving topping the stems and removing the side shoots in Nicotiana rustica, these operations separately and together increased the nicotine content of the leaves.
91. M. C, S. Tm, P. Sr, and M. Dhavaleshvar, “Genetic Diversity Studies on Leaf Yield and Its Component Traits in FCV Tobacco (Nicotiana Tobacum L.) Germplasm Collection,” The Pharma Innovation, vol. 9, no. 11, pp. 346–348, Nov. 2020. doi: 10.22271/tpi.2020.v9.i11f.5377. Abstract
    The present study was aimed to assess the extent of genetic diversity with sixty-six genotypes of FCV tobacco (Nicotiana tobacum L.) by using Mahalanobis’ D² statistics at AINP (Tobacco), Zonal Agriculture and Horticultural Research Station, UAHS, Shivamogga, Karnataka during Kharif-2019-20 in RCBD design with two replications. The 66 genotypes were grouped into eight clusters based on D² analysis. The cluster II had maximum with 29 genotypes followed by clusters I, III, V, and IV had the minimum with 16, 16, 8 and 5 genotypes respectively and remaining clusters were solitary. The highest inter cluster distance was observed between cluster VI and VIII and the lowest between clusters II and VIII. Cluster V had exhibited highest intra cluster distance and the lowest was observed in cluster VI, VII and VII. The character days to flowering, top grade equivalent and green leaf yield showed maximum contribution towards total genetic divergence. On the basis of cluster mean, cluster VI was superior for plant height, number of leaves per plant and leaf length. The maximum leaf width was observed in cluster VIII, while cluster VII sowed superiority for days to flowering. The cluster IV showed highest green leaf yield, cured leaf yield and top grade equivalent. Thus, the genotypes involved in these clusters may be taken into consideration for better parents for generating variability for the respective characters and their rational improvement.
92. D. P. Carew, “Effects of Gibberellic Acid on Nicotiana Rustica Var. Brasilia Plants,” Journal of Pharmaceutical Sciences, vol. 50, no. 12, pp. 1045–1047, Dec. 1961. doi: 10.1002/jps.2600501209. Abstract
    A study was made of the effect of gibberellic acid on the growth of tobacco plants and the accumulation of nicotine. While gibberellic acid increased the rate of growth and the total chlorophyll content, a decrease in nicotine content was observed.
93. N. Chakraborty, M. Banerjee, and K. Acharya, “In Vitro Selection of Elite Clone of Withania Somnifera against Leaf Blight Disease Caused by Alternaria Alternata,” Physiological and Molecular Plant Pathology, vol. 112, p. 101560, Dec. 2020. doi: 10.1016/j.pmpp.2020.101560. Abstract
    In vitro selection of Withania somnifera has been incurred against Alternaria alternata by selection pressure of fungal toxin in an innovative way. Solid green callus was maintained in Murashige and Skoog’s medium supplemented with 6-Benzylaminopurine (2 mgL−1). Toxin was partially purified from A. alternata broth culture. Elite clone was generated by gradual intoxication of solid green callus in the toxin containing medium (5–400 mgL−1). Three highest dose of toxin tolerance was selected viz. 50 mgL−1, 100 mgL−1 and 200 mgL−1. Plants were successfully regenerated from those toxin selected callus. Eventually, various defense enzymes and total phenolics were measured in regenerated plants and toxin selected callus. Callus selected from 100 mgL−1 toxin dose and plants obtained from them give the best results among all and showed more than 2–5 fold increase of defense enzyme activities and total phenolics compared to control. Furthermore, those plants when challenge inoculated showed significant reduction (more than 70%) of leaf spot incidence. Augmentation of disease resistance persists for at least two successive generations. Furthermore, the increase in defense molecules also correlated with increased nitric oxide generation (73%). Presence of withaferin A and withanolide A has also been confirmed in the toxin regenerated plants by High-performance thin-layer chromatography (HPTLC) analysis. Over all observations suggests that fungal toxin at a concentration 100 mgL−1 and by habituation technique can be use as a selection method for the production of disease resistant medicinally important crop like W. somnifera without altering its secondary metabolite production.
94. J. Chandan, S. Gupta, V. Babu, D. Singh, and R. Singh, “Comprehensive Analysis of Codon Usage Pattern in Withania Somnifera and Its Associated Pathogens: Meloidogyne Incognita and Alternaria Alternata,” Genetica, vol. 150, no. 2, pp. 129–144, Apr. 2022. doi: 10.1007/s10709-022-00154-w. Abstract
    Meloidogyne incognita (Root-knot nematode) and Alternaria alternata (fungus) were among the dominant parasites of the medicinal plant Withania somnifera. Despite the fatal nature of their infection, a comprehensive study to explore their evolution and adaptation is lacking. The present study elucidates evolutionary and codon usage bias analysis of W. somnifera (host plant), M. incognita (root-knot nematode) and A. alternata (fungal parasite). The results of the present study revealed a weak codon usage bias prevalent in all the three organisms. Based on the nucleotide analysis, genome of W. somnifera and M. incognita was found to be A-T biased while A. alternata had GC biased genome. We found high similarity of CUB pattern between host and its nematode pathogen as compared to the fungal pathogen. Inclusively, both the evolutionary forces influenced the CUB in host and its associated pathogens. However, neutrality plot indicated the pervasiveness of natural selection on CUB of the host and its pathogens. Correspondence analysis revealed the dominant effect of mutation on CUB of W. somnifera and M. incognita while natural selection was the main force affecting CUB of A. alternata. Taken together the present study would provide some prolific insight into the role of codon usage bias in the adaptability of pathogens to the host’s environment for establishing parasitic relationship.
95. J. J. Chandnani and A. I. Thomas, “Preliminary Studies on Topping and Suckering in Hookah Tobacco (Nicotiana Rustica).,” Indian Journal of Agronomy, vol. 4, pp. 144–7, 1960. https://www.cabdirect.org/cabdirect/abstract/19610304760. Abstract
    An experiment is described which showed that topping and suckering significantly increased leaf area, green weight and weight of cured leaf per plant, and also weight per unit of leaf area. In a second experiment the application of coconut oil after topping to the dormant buds in the top 6 leaf axils suppressed the development of suckers both in number and weight without affecting leaf yield. A...
96. V. kumar Chaturvedi, “A New Method for Grading and Assessing Quality of Motihari Tobacco (Nicotiana Rustica),” Indian Journal of Agricultural Sciences, vol. 60, p. 715, Nov. 1990. Abstract
    In an evaluation of the agmark grading of motihari tobacco (NicoMna TUSticaL.) during 1984, it was found that the criteria for grading are not well defined and the levels of leaf characters of different grades overlap greatly. Also leaf thickness showed no significant correlation with the price of tobacco, indicating that this character is an unsuitable criteria for grading. The price of tobacco was positively correlated with oiliness score, area covered with brown patches (maturityindex),leafweight, leaf area and weight/unit area, but was negatively correlated with areas covered with spots and blemish. Path analysis showed that oiliness score,leafweight and area covered with spots and blemish were the quality determinants of this tobacco. Based on these quality elements, a systematic grading system and a method of assessing quality for this tobacco is proposed. In the agmark grading [Agricultural Produce (Grading and Marketing) Act 1937] of motihan tobacco (Nicotiana rustica L.), leaf thickness, brown patches and per_~ c{~ntage of spots and blemish are the usual criteria for grading. However, drawbacks in this grading system are that leaf thickness is difficult to assess precisely by feel and the levels of leaf thickness and brown patches are not specified in objectively. There is also no clear information about the correlation of these 3 leaf characters (used in the grading system) with the pricing pattern. Hence to clarify these aspects and to develop a method of quality assessment for this tobacco , the present study was conducted. MATERIALS AND METIIODS\vphantom}
97. C. N. Chaubey, S. K. Mishra, and A. P. Mishra, “Study of Variability and Path Analysis for Leaf Yield Components in Hookah Tobacco.,” Tobacco Research, vol. 16, no. 1, pp. 47–52, 1990. https://www.cabdirect.org/cabdirect/abstract/19911620157. Abstract
    Information on yield correlations, heritability and genetic advance is derived from data on 11 yield components in 72 Nicotiana rustica genotypes grown during rabi 1976-77. Leaves/plant, fresh leaf and stalk weight, and days to flowering had high levels of heritability and genetic advance.
98. S. R. Chaudhary, J. Kumar, A. P. Chaudhary, and B. Basak, “Organic Nutrient Sources Influence Yield and Quality of Ashwagandha (Withania Somnifera), and Soil Biochemical Properties,” Agrochimica, vol. 61, no. 3, pp. 221–236, Jul. 2017. doi: 10.12871/00021857201735. Abstract
    A field experiment was conducted to study the effect of organic nutrient management on yield and quality parameters in ashwagandha (Withania somnifera) as well as soil biochemical properties. The experiment was laid out in a split plot design with three replications. The organic manures (farmyard manure, vermicompost and castor cake) were applied as main plot treatments, and biofertilizers and jivamrut were applied in different combination as subplot treatments. The results indicate that the application of organic fertilizers invariably improved root growth, yield and total withanolide content in ashwagandha. The highest dry root yield (714.6 kg ha⁻¹) and total withanolide content (0.91 mg g⁻¹) were recorded with application of castor cake, which was at par with vermicompost. Soil available nutrients and biochemical properties were significantly improved under vermicompost and castor cake applications as compared to other treatments. Thus, organic manures, particularly castor cake and vermicompost, could be a promising source of nutrients in organic cultivation of ashwagandha.
99. S. Chaurasia and R. Singh, “Effect of Integrated Nutrient Management System on Yield Attributes, Yield and Economics of Ashwagandha (Withania Somnifera (l,” International Journal of Current Science, vol. 12, no. 2, p. 20, 2022. Abstract
    Ashwagandha (Withania somnifera (L.) Dunal) is one of the most important medicinal crop in India. It plays an important role in medicinal industries. The demand of fleshy whitish brown roots are high but the production is low. Therefore, the present study was conducted to know the effect of integrated nutrient management system on yield attributes, yield and economics of Ashwagandha (Withania somnifera (L.) Dunal) at near the college of agriculture farm, Tikamgarh (M.P.) India during kharif season of 2017-18 and 2018-19. The experiment was laid out in randomized block design with three replications and twelve treatments viz, 100% NPK recommended dose 50:30:30 kg/ha (T1), 100% NPK/ha + 5 kg Zn/ha (T2), 10 tonnes FYM/ha (T3), 10 tonnes FYM/ha + 3 kg PSB/ha (T4), 10 tonnes FYM/ha + 3 kg Azotobacter/ha (T5), 10 tonnes FYM/ha + 3 kg Azotobacter/ha + 3 kg PSB/ha (T6), 10 tonnes FYM/ha + 3 kg Azotobacter/ha + 3 kg PSB/ha + 5 kg Zn/ha (T7), 50% NPK/ha + 5 tonnes FYM/ha (T8), 50% NPK/ha + 5 tonnes FYM/ha + 3 kg PSB/ha (T9), 50% NPK/ha + 5 tonnes FYM/ha + 3 kg Azotobacter/ha (T10), 50% NPK/ha + 5 tonnes FYM/ha + 3 kg Azotobacter/ha + 3 kg PSB/ha (T11) and 50% NPK/ha + 5 tonnes FYM/ha + 3 kg Azotobacter/ha + 3 kg PSB/ha + 5 kg Zn/ha (T12). The pooled analysis of two years data indicated that an integrated nutrient management system significantly influenced different yield attributes, yield and economic parameters of Ashwagandha. The significant maximum values were recorded in number of berries per plant (121.61), number of seeds par berry (40.54), 1000 seed weight (4.46 g), root yield per plant (39.93 g), shoot yield per plant (115.80 g), seed yield per plant (12.80 g), root yield per hectare (629.69 kg/ha), gross returns (Rs. 1,20,881/ha), net returns (Rs. 81,887/ha) and benefit cost ratio (3.10) with 50% NPK/ha + 5 tonnes FYM/ha + 3 kg Azotobacter/ha + 3 kg PSB/ha + 5 kg Zn/ha (T12) treatments. While highest seed yield per hectare (175.30 kg/ha) was recorded with the application of 100% NPK recommended dose 50:30:30 kg/ha (T1). Based on overall experimental results, it could be concluded that 50% NPK/ha + 5 tonnes FYM/ha + 3 kg Azotobacter/ha + 3 kg PSB/ha + 5 kg Zn/ha (T12) treatment was found to be a better integrated nutrient management system which can be used for better quality production, higher yield and maximum economic benefit of Ashwagandha (Withania somnifera (L.) Dunal) crop.
100. N. D. Chaurasiya, N. S. Sangwan, F. Sabir, L. Misra, and R. S. Sangwan, “Withanolide Biosynthesis Recruits Both Mevalonate and DOXP Pathways of Isoprenogenesis in Ashwagandha Withania Somnifera L. (Dunal),” Plant Cell Reports, vol. 31, no. 10, pp. 1889–1897, Oct. 2012. doi: 10.1007/s00299-012-1302-4. Abstract
     Withanolides are pharmaceutically important C28-phytochemicals produced in most prodigal amounts and diversified forms by Withania somnifera. Metabolic origin of withanolides from triterpenoid pathway intermediates implies that isoprenogenesis could significantly govern withanolide production. In plants, isoprenogenesis occurs via two routes: mevalonate (MVA) pathway in cytosol and non-mevalonate or DOXP/MEP pathway in plastids. We have investigated relative carbon contribution of MVA and DOXP pathways to withanolide biosynthesis in W. somnifera. The quantitative NMR-based biosynthetic study involved tracing of 13C label from 13C1-d-glucose to withaferin A in withanolide producing in vitro microshoot cultures of the plant. Enrichment of 13C abundance at each carbon of withaferin A from 13C1-glucose-fed cultures was monitored by normalization and integration of NMR signal intensities. The pattern of carbon position-specific 13C enrichment of withaferin A was analyzed by a retro-biosynthetic approach using a squalene-intermediated metabolic model of withanolide (withaferin A) biosynthesis. The pattern suggested that both DOXP and MVA pathways of isoprenogenesis were significantly involved in withanolide biosynthesis with their relative contribution on the ratio of 25:75, respectively. The results have been discussed in a new conceptual line of biosynthetic load-driven model of relative recruitment of DOXP and MVA pathways for biosynthesis of isoprenoids.
101. M. Chavan, “Influence of Dates of Sowing and Plant Density on Root Morphology and Root Yield of Ashwagandha (Withania Somnifera),” Journal of Ecobiology, vol. 26, no. 1, pp. 285–292, 2010. https://www.academia.edu/12503503/INFLUENCE_OF_DATES_OF_SOWING_AND_PLANT_DENSITY_ON_ROOT_MORPHOLOGY_AND_ROOT_YIELD_OF_ASHWAGANDHA_Withania_somnifera_. Abstract
     The research study was conducted to know the influence of dates of sowing on ashwagandha (Withania somnifera Dunal) at Arabhavi during 2006 with factorial randomized block design. To study the effect of sowing dates I and II fortnights of July and
102. D. Chitturi, R. K. Venisetty, R. K. Molmoori, C. K. Kokate, and S. S. Apte, “Enhanced Bioproduction of Withaferin A from Suspension Cultures of Withania Somnifera.,” Annals of Biological Research, vol. 1, no. 2, pp. 77–86, 2010. https://www.cabdirect.org/cabdirect/abstract/20103256568. Abstract
     Callus cultures of Withania somnifera from leaves were established on MS (Murashige and Skoog) media supplemented with Dicamba (2 mg/l), Kinetin (0.1 mg/l) and Sucrose (3% w/v). Suspension cultures were established and the growth and production kinetics were studied. For growth kinetics MS media supplemented with Dicamba (2 mg/l), Kinetin (0.1 mg/l) and Sucrose (3% w/v) without agar was used...
103. D. Choudhary, “Causes of Poor and Erratic Germination in Atropa Belladonna,” Planta Medica, vol. 27, no. 01, pp. 18–22, Feb. 1975. doi: 10.1055/s-0028-1097754. Abstract
     An effort has been made to arzalyse the probable causes of a low germination o f Atropa species. From the results achieved it is expected that the very hard, thick, woody and waxy nature of testa along with an internal membrane enclosing the embryo and the presence .of seeds containing only a rudimentary embryo may be the va/ious factors responsible for poor and erratic gernlination of Atropa species.
104. M. K. U. Chowdhury, “A Comparative Study of Genotypic and Environmental Response to Androgenesis in Nicotiana Rustica,” Theoretical and Applied Genetics, vol. 70, no. 2, pp. 128–132, May 1985. doi: 10.1007/BF00275311. Abstract
     A total of six genotypes of Nicotiana rustica comprising the two F1’s (V2 × V12 and V1 × V5) and their parents were evaluated for their efficiency in haploid production. Excised immature flower buds with pollen at late uninucleate to early binucleate stage were pretreated for 21 days at 5 ° or 7 °C, or for 15 days at 9 °C before culturing on Nitsch’s medium+ 0.1 mg/l NAA. The effects of genotype, pretreatment and their interaction were tested on anther response, anther productivity and days to first plantlet formation. Highly significant genotype X pretreatment interaction and differences between genotypes were observed for all three characters. Significant differences between pretreatments were observed for anther productivity only. The performance of V12 both in respect of anther productivity and response was highest whereas that of V5 was the lowest. Analysis of variance showed that a simple additive genetic model was not adequate to explain the above variation due to significant additive genetic and dominance interactions with the pretreatment.
105. M. K. U. Chowdhury, “An Improved Method for Dihaploid Production in Nicotiana Rustica through Anther Culture,” Theoretical and Applied Genetics, vol. 69, no. 2, pp. 199–204, Dec. 1984. doi: 10.1007/BF00272895. Abstract
     The efficiency of dihaploid production from anther culture in N. rustica has been improved by studying the effects of pretreatment temperature, pretreatment duration and initial anther stage on anther response, anther productivity and time to first plantlet production. Pretreatment was most effective on anthers at or around the stage of pollen mitosis. Pollen mitosis stage anthers pretreated at 9 °C for 15 days gave the best results. Both spontaneous and induced dihaploids were obtained. Small plantlets treated with 0.4% colchicine and 2% DMS solution for 5 h produced the maximum number of dihaploids (more than 50%). These considerable improvements in the efficiency of the techniques have made dihaploidy an attractive method for producing inbred lines in N. rustica. This will permit a large scale comparison of dihaploids with more conventional methods of inbreeding such as single seed descent and pedigree breeding.
106. P. Choyal et al., “Chemical Manipulation of Source and Sink Dynamics Improves Significantly the Root Biomass and the Withanolides Yield in Withania Somnifera,” Industrial Crops and Products, vol. 188, p. 115577, Nov. 2022. doi: 10.1016/j.indcrop.2022.115577. Abstract
     Roots are the main economic part of Withania somnifera having high commercial value in the pharmaceutical industry. Since W. somnifera has the indeterminant type of growth habit, it exhibits a continuous berry formation which ultimately reduces the root yield. The present study was undertaken to determine the effect of altered source - sink relationship by regulating the sink availability to increase the root yields. Maleic hydrazide (MH @ 200, 500 and 1000 μmol mol−1) and Gibberellic acid (GA @ 500 and 1000 μmol mol−1) were used as chemical gametocide for inducing male sterility while mechanical deflowering was used to prevent the berry formation. In all three approaches, the root biomass and withanolides content, in general, were significantly increased. The dry matter partitioning pattern showed that only 6% of total biomass was accumulated in the roots whereas 37% accululated in the shoot and a significant amount i.e., 57% was partitioned to the berries under control condition. An application of MH and GA and the mechanical deflowering treatment, however, changed this pattern in favour of the roots. As much as 26% of the total biomass was partitioned to the roots under the MH treatments. Whereas, GA (500 μmol mol−1) and mechanical deflowering treatment increased the root biomass by 75 and more than 300% respectively. More than 200% increase in withaferin A and ∼100% increase in withanolide A content was observed in leaves under the GA (500 μmol mol−1) treatment. Mechanical deflowering treatment was also equally efficient as GA in increasing the withanolides both in the leaves and the roots. The photosynthetic traits, total sugars and starch content, and sugar metabolizing enzyme analysis showed that modification in source-sink relationship through inhibited berry formation triggers a higher partitioning of carbon assimilates from source to the roots and thus improves significantly the root biomass and the withanolide yields in W. somnifera. The same was validated through the radiotracer studies involving 14C.
107. V. Ciddi, “Withaferin A from Cell Cultures of Withania Somnifera,” Indian Journal of Pharmaceutical Sciences, vol. 68, no. 4, 2006. doi: 10.4103/0250-474X.27824. Abstract
     Suspension cultures of Withania somnifera cells were established and shown to produce withaferin A. The identification of withaferin A was done by TLC, UV absorption, HPLC and electron spray mass spectroscopy. These cultures could be strongly elicited by exposure to salacin. Addition of salacin at the concentration of 750 µM to the cultures in production medium enhanced production levels of withaferin A to 25±2.9 mg/l compared to 0.47±0.03 mg/l in unelicited controls. This report is the first to demonstrate withaferin A production in plant suspension cultures and provides prerequisites for commercial scale, controlled production of withaferin A.
108. J. J. Clarkson, S. Dodsworth, and M. W. Chase, “Time-Calibrated Phylogenetic Trees Establish a Lag between Polyploidisation and Diversification in Nicotiana (Solanaceae),” Plant Systematics and Evolution, vol. 303, no. 8, pp. 1001–1012, Oct. 2017. doi: 10.1007/s00606-017-1416-9. Abstract
     We investigate the timing of diversification in allopolyploids of Nicotiana (Solanaceae) utilising sequence data of maternal and paternal origin to look for evidence of a lag phase during which diploidisation took place. Bayesian relaxed clock phylogenetic methods show recent allopolyploids are a result of several unique polyploidisation events, and older allopolyploid sections have undergone subsequent speciation at the polyploid level (i.e. a number of these polyploid species share a singular origin). The independently formed recent polyploid species in the genus all have mean age estimates below 1 million years ago (Ma). Nicotiana section Polydicliae (two species) evolved 1.5 Ma, N. section Repandae (four species) formed 4 Ma, and N. section Suaveolentes (~35 species) is about 6 million years old. A general trend of higher speciation rates in older polyploids is evident, but diversification dramatically increases at approximately 6 Ma (in section Suaveolentes). Nicotiana sect. Suaveolentes has spectacularly radiated to form 35 species in Australia and some Pacific islands following a lag phase of almost 6 million years. Species have filled new ecological niches and undergone extensive diploidisation (e.g. chromosome fusions bringing the ancestral allotetraploid number, n = 24, down to n = 15 and ribosomal loci numbers back to diploid condition). Considering the progenitors of Suaveolentes inhabit South America, this represents the colonisation of Australia by polyploids that have subsequently undergone a recent radiation into new environments. To our knowledge, this study is the first report of a substantial lag phase being investigated below the family level.
109. M. Condorí, F. Albesa, F. Altobelli, G. Duran, and C. Sorrentino, “Image Processing for Monitoring of the Cured Tobacco Process in a Bulk-Curing Stove,” Computers and Electronics in Agriculture, vol. 168, p. 105113, Jan. 2020. doi: 10.1016/j.compag.2019.105113. Abstract
     A control system based on digital image processing was developed for the tobacco curing process in a bulk-curing barn. A laboratory prototype of the barn was built to test the automation of the process and the results with virginia type tobacco curing are presented. The system records in real-time dry-bulb temperature, air relative humidity, the weight loss of a sample of tobacco leaves and the image of a tobacco leaf. A value for the air humidity ratio of 0.045 kg/kg through the process is obtained as the optimal curing condition. A finite state machine was developed to control the curing condition through the image processing. The HSV color system was used in the image processing; the optimum color for the yellowing stage was obtained with 53.4 for the channel H. In this stage, the time derivative of H close to zero was used as the condition of color stability. The time derivative close to zero for the standard deviation of S was used as uniformity color condition in the color fixing stage and in the leaf drying stage. A good fit between the weight loss and the channel H was obtained; because of this, it was used as the indicator to follow the weight loss in leaves.
110. V. C. Cooper and D. G. A. Walkey, “Thermal Inactivation of Cherry Leaf Roll Virus in Tissue Cultures of Nicotiana Rustica Raised from Seeds and Meristem-Tips,” Annals of Applied Biology, vol. 88, no. 2, pp. 273–278, 1978. doi: 10.1111/j.1744-7348.1978.tb00706.x. Abstract
     Nicotiana rustica tissue cultures derived from seeds or embryos infected with cherry leaf roll virus (CLRV), remained infected after culture at 22 oC. No infectivity was found in cultures held at 32 oC for 5 days but it was readily detected after such cultures were transferred to 25 oC for 8 days. Virus was permanently eradicated from most plants after 20 days incubation at 32 oC and from all plants after 7 days incubation at 40 oC. Partially purified preparations of CLRV lost infectivity after 9–12 days at 22oC, 5 days at 32oC and 3 days at 40oC.
111. C. çrak, K. Kevseroglu, and B. Sağlam, “Physical and Physiological Dormancy in Black Henbane (Hyoscyamus Niger L.) Seeds,” Journal of Plant Biology, vol. 47, no. 4, pp. 391–395, Dec. 2004. doi: 10.1007/BF03030556. Abstract
     Aim of this study was to investigate the nature of dormancy in black henbane (Hyoscyamus niger) seeds which have low germination rate under normal laboratory conditions. To do this, before placing the seeds in Petri dishes, they were soaked in 5,10 and 15 mg/L GA; 1,2 and 3% H2SO4, 15 mg/L GA + 1% H2SO4, 0.01 M KNO3 solutions, tap water, 40, 50 and 60°C hot water for 30 min. The study was performed under both continuous illumination and darkness in growth chambers to evaluate the effect of light on germination rate. The results showed that H2SO4 and GA treatments were the most important factors affecting seed germination and their germination enhancing effects were more evident in darkness. The results also suggested that black henbane seeds exhibit double dormancy involving a hard seed coat and a partially dormant embryo and have a partial dark requirement to germinate.
112. B. T. Cromwell, “Experiments on the Synthesis of Hyoṡcyamine in Atropa Belladonna,” Biochemical Journal, vol. 31, no. 4, pp. 551–559, Apr. 1937. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1266974/.
113. B. T. Cromwell, “Studies on the Synthesis of Hyoscyamine in Atropa Belladonna L. and Datura Stramonium L,” Biochemical Journal, vol. 37, no. 6, pp. 717–722, 1943. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1258005/.
114. A. J. Das and R. Kumar, “Bioremediation of Petroleum Contaminated Soil to Combat Toxicity on Withania Somnifera through Seed Priming with Biosurfactant Producing Plant Growth Promoting Rhizobacteria,” Journal of Environmental Management, vol. 174, pp. 79–86, Jun. 2016. doi: 10.1016/j.jenvman.2016.01.031. Abstract
     Soil contaminated by Petroleum oil cannot be utilized for agricultural purposes due to hydrocarbon toxicity. Oil contaminated soil induces toxicity affecting germination, growth and productivity. Several technologies have been proposed for bioremediation of oil contaminated sites, but remediation through biosurfactant producing plant growth promontory rhizobacteria (PGPR) is considered to be most promising methods. In the present study the efficacy of seed priming on growth and pigment of Withania somnifera under petroleum toxicity is explored. Seeds of W. somnifera were primed with biosurfactant producing Pseudomonas sp. AJ15 with plant growth promoting traits having potentiality to utilized petroleum as carbon source. Results indicates that plant arose from priming seeds under various petroleum concentration expressed high values for all the parameters studied namely germination, shoot length, root length, fresh and dry weight and pigments (chlorophyll and carotenoid) as compared to non primed seed. Hence, the present study signifies that petroleum degrarding biosurfactant producing PGPR could be further used for management and detoxification of petroleum contaminated soils for growing economically important crops.
115. M. G. Dasgupta, B. S. George, A. Bhatia, and O. P. Sidhu, “Characterization of Withania Somnifera Leaf Transcriptome and Expression Analysis of Pathogenesis – Related Genes during Salicylic Acid Signaling,” PLOS ONE, vol. 9, no. 4, p. e94803, Apr. 2014. doi: 10.1371/journal.pone.0094803. Abstract
     Withania somnifera (L.) Dunal is a valued medicinal plant with pharmaceutical applications. The present study was undertaken to analyze the salicylic acid induced leaf transcriptome of W. somnifera. A total of 45.6 million reads were generated and the de novo assembly yielded 73,523 transcript contig with average transcript contig length of 1620 bp. A total of 71,062 transcripts were annotated and 53,424 of them were assigned GO terms. Mapping of transcript contigs to biological pathways revealed presence of 182 pathways. Seventeen genes representing 12 pathogenesis-related (PR) families were mined from the transcriptome data and their pattern of expression post 17 and 36 hours of salicylic acid treatment was documented. The analysis revealed significant up-regulation of all families of PR genes by 36 hours post treatment except WsPR10. The relative fold expression of transcripts ranged from 1 fold to 6,532 fold. The two families of peroxidases including the lignin-forming anionic peroxidase (WsL-PRX) and suberization-associated anionic peroxidase (WsS-PRX) recorded maximum expression of 377 fold and 6532 fold respectively, while the expression of WsPR10 was down-regulated by 14 fold. Additionally, the most stable reference gene for normalization of qRT-PCR data was also identified. The effect of SA on the accumulation of major secondary metabolites of W. somnifera including withanoside V, withaferin A and withanolide A was also analyzed and an increase in content of all the three metabolites were detected. This is the first report on expression patterns of PR genes during salicylic acid signaling in W. somnifera.
116. A. K. Datta, A. Das, A. Bhattacharya, S. Mukherjee, and B. K. Ghosh, “An Overview on Withania Somnifera (L.) Dunal – The ‘Indian Ginseng,’” Medicinal and Aromatic Plant Science and Biotechnology, vol. 5, no. 1, pp. 1–15, 2011. https://www.academia.edu/72120219/An_Overview_on_Withania_somnifera_L_Dunal_The_Indian_ginseng_. Abstract
     Withania somnifera (L.) Dunal (Family: Solanaceae; commonly known as Ashwagandha; English name, winter cherry) is a perennial plant species with profound therapeutic significance in both traditional (Ayurvedic, Unani, Sidhdha) and modern systems of medicine. Due to the restorative property of roots, the species is also known as ‘Indian ginseng’. With a view to the medicinal importance of the species an overview is conducted involving nearly all essential aspects to provide updated, adequate information to researchers for effective utilization in human benefit.
117. R. F. Dawson and M. L. Solt, “Estimated Contributions of Root and Shoot to the Nicotine Content of the Tobacco Plant. 1,” Plant Physiology, vol. 34, no. 6, pp. 656–661, Nov. 1959. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC541275/.
118. S. M. Dehdashti, S. Acharjee, S. Kianamiri, and M. Deka, “An Efficient Agrobacterium Rhizogenes-Mediated Transformation Protocol of Withania Somnifera,” Plant Cell, Tissue and Organ Culture (PCTOC), vol. 128, no. 1, pp. 55–65, Jan. 2017. doi: 10.1007/s11240-016-1081-7. Abstract
     This is the first report on Agrobacterium rhizogenes-mediated transformation of Withania somnifera for expression of a foreign gene in hairy roots. We transformed leaf and shoot tip explants using binary vector having gusA as a reporter gene and nptII as a selectable marker gene. To improve the transformation efficiency, acetosyringone (AS) was added in three stages, Agrobacterium liquid culture, Agrobacterium infection and co-culture of explants with Agrobacterium. The addition of 75 μM AS to Agrobacterium liquid culture was found to be optimum for induction of vir genes. Moreover, the gusA gene expression in hairy roots was found to be best when the leaves and shoot tips were sonicated for 10 and 20s, respectively. Based on transformation efficiency, the Agrobacterium infection for 60 and 120 min was found to be suitable for leaves and shoot tips, respectively. Amongst the various culture media tested, MS basal medium was found to be best in hairy roots. The transformation efficiency of the improved protocol was recorded 66.5 and 59.5\,% in the case of leaf and shoot tip explants, respectively. When compared with other protocols the transformation efficiency of this improved protocol was found to be 2.5 fold higher for leaves and 3.7 fold more for shoot tips. Southern blot analyses confirmed 1–2 copies of the gusA transgene in the lines W1-W4, while 1–4 transgene copies were detected in the line W5 generated by the improved protocol. Thus, we have established a robust and efficient A. rhizogenes mediated expression of transgene (s) in hairy roots of W. somnifera.
119. P. U. Devi, “Withania Somnifera Dunal (Ashwagandha): Potential Plant Source of a Promising Drug for Cancer Chemotherapy and Radiosensitization,” Indian journal of experimental biology, vol. 34, no. 10, pp. 927–932, Oct. 1996. Abstract
     Study of antitumor and radiosensitizing properties of W. somnifera (Ashwagandha), a well known medicinal plant, have yielded encouraging results. The alcoholic extract of the dried roots of the plant as well as the active component withaferin A isolated from the extract showed significant antitumor and radiosensitizing effects in experimental tumors in vivo, without any noticeable systemic toxicity. Withaferin A gave a sensitizer enhancement ratio of 1.5 for in vitro cell killing of V79 Chinese hamster cells at a non toxic concentration of approximately 2 microM. The mechanism of action of this compound is not known. The studies so far indicate that W. somnifera could prove to be a good natural source of a potent and relatively safe radiosensitizer/chemotherapeutic agent. Further studies are needed to explore the clinical potential of this plant for cancer therapy.
120. S. T. Devkar, S. D. Jagtap, S. S. Katyare, and M. V. Hegde, “Estimation of Antioxidant Potential of Individual Components Present in Complex Mixture of Withania Somnifera (Ashwagandha) Root Fraction by Thin-Layer Chromatography—2,2-Diphenyl-1-Picrylhdrazyl Method,” JPC – Journal of Planar Chromatography – Modern TLC, vol. 27, no. 3, pp. 157–161, Jun. 2014. doi: 10.1556/JPC.27.2014.3.2. Abstract
     A thin-layer chromatography–2,2-diphenyl-1-picrylhdrazyl (TLC—DPPH) method has been developed for the quantification of withanolides and simultaneous estimation of antioxidant potential of each constituent present in a complex mixture. High-performance thin-layer chromatography (HPTLC) of withanolide-rich fraction of Withania somnifera root was performed on Si 60F254 HPTLC plates with dichloromethane-toluene-methanol-acetone-diethyl ether (6.5:7:4:1.5:1, v/v) as a mobile phase for the separation of withanolides and on the basis of RF values 0.07, 0.55, 0.61, 0.63, 0.66, and 0.77 for withanoside V, withaferine A, 1,2-deoxywithastramonolide, withanone, withanolide A, and withanolide B, respectively. The concentrations of withanoside V, withaferine A, 1,2-deoxywithastramonolide, withanone, withanolide A, and withanolide B were 0.86, 12.9, 1.92, 1.52, 5.24, and 4.52 mg g−1 respectively. A TLC-DPPH rapid test was used to assess the free-radical scavenging activity of constituents in withanolide-rich fraction for the first time. In withanolides, withanoside V and withanolide B show the highest antioxidant potential. In unknown compounds, peak No. 17 at RF 0.92 has the highest antioxidant potential and peak No. 16 at RF 0.87 has recorded the lowest antioxidant potential. It is also found that some of the unidentified components had significant antioxidant potential. This technique has several advantages; therefore, this method will be useful for rapidly validating the Ayurvedic products containing W. somnifera.
121. S. Devkar, Y. Badhe, S. Jagtap, and M. Hegde, “Quantification of Major Bioactive Withanolides in Withania Somnifera (Ashwagandha) Roots by HPTLC for Rapid Validation of Ayurvedic Products,” JPC - Journal of Planar Chromatography - Modern TLC, vol. 25, no. 4, pp. 290–294, Jul. 2012. doi: 10.1556/jpc.25.2012.4.2. Abstract
     The high-performance thin-layer chromatography (HPTLC) method has been developed for the simultaneous quantification of withaferine A, 1,2 deoxy-withastramonolide, withanolide A, and withanolide B for the validation of Withania somnifera (Ashwagandha roots) as raw material and Ashwagandha-containing finished Ayurvedic products. HPTLC of W. somnifera methanolic extracts was performed on Si 60F254 (10 × 10 cm) HPTLC plates with dichloromethane-toluene-methanol-acetone-diethyl ether (7.5:7.5: 3:1:1 v/v) as a mobile phase. Upon separation, quantitative evaluation of these withanolides was performed in the absorption reflection mode at 235 nm. The method was validated for precision, reproducibility, and accuracy. On the basis of RF values of 0.58, 0.61, 0.68, and 0.79 for withaferine A, 1,2 deoxy-withastramonolide, withanolide A, and withanolide B, respectively, were identified. On the basis of linear calibration curves for all withanolides in the range of 0.2-1.2 μg, an average recovery of withaferine A, 1,2 deoxy-withastramonolide, withanolide A, and withanolide B was 98%, 99.5%, 98%, and 99%, respectively. The method is very simple, precise, specific, sensitive, accurate, and economical for rapidly validating the Ayurvedic products containing W. somnifera (Ashwagandha).
122. Y. H. Dewir, D. Chakrabarty, S.-H. Lee, E.-J. Hahn, and K.-Y. Paek, “Indirect Regeneration of Withania Somnifera and Comparative Analysis of Withanolides in in Vitro and Greenhouse Grown Plants,” Biologia Plantarum, vol. 54, no. 2, pp. 357–360, Jun. 2010. doi: 10.1007/s10535-010-0063-6. Abstract
     The present study reports an efficient protocol for indirect shoot organogenesis and plantlets regeneration of Withania somnifera (L.) Dunal. Leaf explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of 6-benzylaminopurine (BAP) and indole-3-acetic acid (IAA). The highest callus induction rate (89.5 %) and shoot regeneration rate (92 %) were obtained when 2 mg dm−3 BAP was combined with 0.5 mg dm−3 IAA. Three major withanolides (withaferine A, 12-deoxywithastramonolide and withanolide A) were investigated in different plant organs from in vitro and greenhouse grown plants. Leaves contained higher contents of withanolides and phenolics than roots or stems, whereas roots contained the highest contents of flavonoids and polysacharides. In vitro grown plants contained greater contents of phenolics, flavonoids and polysaccharides while lower contents of withanolides than greenhouse grown plants.
123. N. S. Dhalla, M. S. Sastry, and C. L. Malhotra, “Chemical Studies of the Leaves of Withania Somnifera,” Journal of Pharmaceutical Sciences, vol. 50, no. 10, pp. 876–877, Oct. 1961. doi: 10.1002/jps.2600501019. Abstract
     Besides the preliminary investigations, a new crystalline compound named withanone (m. p. 263 °) has been isolated from the leaves of Withania somnifera. Withanone is found to contain lactone and steroidal rings in the molecule. Somnitol, glucose, and a large quantity of inorganic salts have also been separated. The total alkaloids, on subjecting to paper chromatography, indicate five alkaloidal bands. The presence of cystine, glycine, glutamic acid, α-alanine, proline, and tryptophan is also reported.
124. N. Dhar et al., “Cloning and Functional Characterization of Three Branch Point Oxidosqualene Cyclases from Withania Somnifera (L.) Dunal *,” Journal of Biological Chemistry, vol. 289, no. 24, pp. 17249–17267, Jun. 2014. doi: 10.1074/jbc.M114.571919. Abstract

     Oxidosqualene cyclases (OSCs) positioned at a key metabolic subdividing junction execute indispensable enzymatic cyclization of 2,3-oxidosqualene for varied triterpenoid biosynthesis. Such branch points present favorable gene targets for redirecting metabolic flux toward specific secondary metabolites. However, detailed information regarding the candidate OSCs covering different branches and their regulation is necessary for the desired genetic manipulation. The aim of the present study, therefore, was to characterize members of OSC superfamily from Withania somnifera (Ws), a medicinal plant of immense repute known to synthesize a large array of biologically active steroidal lactone triterpenoids called withanolides. Three full-length OSC cDNAs, β-amyrin synthase (WsOSC/BS), lupeol synthase (WsOSC/LS), and cycloartenol synthase (WsOSC/CS), having open reading frames of 2289, 2268, and 2277 bp, were isolated. Heterologous expression in Schizosaccharomyces pombe, LC-MS analyses, and kinetic studies confirmed their monofunctionality. The three WsOSCs were found to be spatially regulated at transcriptional level with WsOSC/CS being maximally expressed in leaf tissue. Promoter analysis of three WsOSCs genes resulted in identification of distinct cis-regulatory elements. Further, transcript profiling under methyl jasmonate, gibberellic acid, and yeast extract elicitations displayed differential transcriptional regulation of each of the OSCs. Changes were also observed in mRNA levels under elicitations and further substantiated with protein expression levels by Western blotting. Negative regulation by yeast extract resulted in significant increase in withanolide content. Empirical evidence suggests that repression of competitive branch OSCs like WsOSC/BS and WsOSC/LS possibly leads to diversion of substrate pool toward WsOSC/CS for increased withanolide production.

125. K. Dharmar and A. J. D. Britto, “RAPD Analysis of Genetic Variability in Wild Populations of Withania Somnifera (L.) Dunal,” p. 5, 2011. Abstract
     Withania somnifera is extensively used as herbal medicine, because it contains clinically important compounds. In the present study, the genetic variability in W. somnifera among accessions of different geographical region in Tamil Nadu was assessed through Random Amplified Polymorphic DNA (RAPD) markers. Five accessions of W. somnifera were screened with ten primers of which six primers were found to be the most informative. These primers produced multiple band profiles with a number of amplified DNA fragments varying from 5 to 9. A total of 37 polymorphic bands were observed. The genetic distance between the population ranged from 0.2436 to 0.4754 and the genetic identity ranged from 0.6216 to 0.7838. The overall observed and effective number of alleles was about 1.6216 and 1.4271 respectively. Nei’s over all genetic diversity is 0.2465. The degree of percentage of polymorphism (83.78 %) was found to be high in accession collected from Samykoundanpalayam in Tirupur District, Tamil Nadu. The information obtained here could be valuable for devising strategies for conservation this medicinal plant.
126. M. Dijak, A. Sproule, W. Keller, R. Pandeya, and S. Gleddie, “Transformation of Nicotiana Tabacum, N. Debneyi, and N. Rustica: Inheritance and Protoplast Expression of Antibiotic Resistance,” Plant Cell, Tissue and Organ Culture, vol. 25, no. 3, pp. 189–197, Jun. 1991. doi: 10.1007/BF00036210. Abstract
     Agrobacterium tumefaciens strains harbouring plasmid vectors pBCAT1, pVU1011 or pMON806 were used to transform leaf explants of Nicotiana tabacum cultivars ‘Delgold’ and ‘Candel’, N. debneyi, and N. rustica var. NRT. Transgenic plants resistant to the selective agents kanamycin, hygromycin or methotrexate were regenerated and used as sources of leaf mesophyll protoplasts. Protoplasts divided and regenerated plants in the presence of selective agents at levels inhibitory to protoplasts of non-transformed plants. Cross-resistance of protoplasts to more than one selective agent was not observed in this study which suggests that this approach may lead to an efficient interspecific somatic hybrid selection system.
127. D. Dikasso et al., “Anti-Malarial Activity of Withania Somnifera L. Dunal Extracts in Mice,” Ethiopian medical journal, vol. 44, no. 3, pp. 279–285, Jul. 2006. Abstract
     ObjectiveThis study aims at investigating the in vivo antiplasmodial activity of a traditionally used medicinal plant, Withania somnifera, L. Dunal, (Solanaceae).MethodsRodent malaria parasite, Plasmodium berghei, 0.2 ml of x l0(7) parasites, was inoculated into Swiss albino mice intraperitoneally. Extracts were administered by intra gastric tube daily for four days starting from the day of parasite inoculation. Negative controls received the same amount of solvent used to suspend the exracts and the positive controls were given chloroquine by the same route. Parasite suppressive effect and effects on body weight, packed cell volume (PCV) and body temperature were determined.ResultsParasitemia percent inhibition of W. somnifera roots and root barks were 50.43% and 29.13% respectively, with 600 mg/kg dose. Inhibition was statistically significant at all dose levels, compared to the negative controls (p ConclusionExtracts of the leaves and root barks of W. somnifera showed parasite suppressive effect and a protective effect on PCV drop (at higher doses), both in dose-related fashions. However, the effects on body weight and body temperature falls are inconclusive.
128. R. Dimitrijevic, “The effect of different combinations and rates of mineral fertilizers on the leaf yield and nicotine content of Nicotiana rustica L.,” The effect of different combinations and rates of mineral fertilizers on the leaf yield and nicotine content of Nicotiana rustica L., no. 291, 1960. https://www.cabdirect.org/cabdirect/abstract/19621902429. Abstract
     Highest concentrations and yields of nicotine were produced with dressings of 50 kg/ha each of N and P and 75 kg K, or 150 kg each of N and K, or 150 kg N and 225 kg P. Further increase in application rates did not produce proportional increases in yields.
129. G. C. Douglas, L. R. Wetter, C. Nakamura, W. A. Keller, and G. Setterfield, “Somatic Hybridization between Nicotiana Rustica and N. Tabacum. III. Biochemical, Morphological, and Cytological Analysis of Somatic Hybrids,” Canadian Journal of Botany, vol. 59, no. 2, pp. 228–237, Feb. 1981. doi: 10.1139/b81-034.
130. G. C. Douglas, W. A. Keller, and G. Setterfield, “Somatic Hybridization between Nicotiana Rustica and N. Tabacum. I. Isolation and Culture of Protoplasts and Regeneration of Plants from Cell Cultures of Wild-Type and Chlorophyll-Deficient Strains,” Canadian Journal of Botany, vol. 59, no. 2, pp. 208–219, Feb. 1981. doi: 10.1139/b81-032. Abstract
     Preliminary to somatic hybridization between chlorophyll-deficient mutants of Nicotiana rustica and N. tabacum conditions for cell culture, protoplast production and culture and induction of morphogenesis were established. Callus and suspension cultures from wild-type and chlorophyll-deficient strains of both species were established and maintained on Murashige and Skoog medium containing either 2,4-dichlorophenoxyacetic acid (2,4-D) or naphthaleneacetic acid (NAA). The chlorophyll-deficient phenotype was evident in cultures derived from mutants of both species. Protoplasts were readily obtained from cell cultures with N. rustica outyielding N. tabacum and mutant cells of the latter substantially outyielding the wild-type cells. In Nagata and Takebe medium protoplasts regenerated cells walls and 60–80% divided and formed cell colonies. Protoplasts from cultures on 2,4-D medium were healthier but slower to divide than those from cultures grown on an NAA-supplemented medium. Protoplasts derived from 2,4-D-grown cells also gave rise to proembryo-like structures. Shoot and root organogenesis could be induced in cell colonies obtained from both suspension cultures and protoplasts of mutant and wild-type N. rustica. With albino N. tabacum only cell colonies from cell suspension showed morphogenic potential. Maximum frequencies of colonies forming shoots were 77% for N. rustica cv. chlorotica and 100% for albino N. tabacum. The morphogenic capacity of colonies from 2,4-D-grown cells was greater than from cells grown on NAA. Morphogenic capacity of both species declined with increasing age of the suspension cultures. Roots were induced on shoots of the wild type in both species as well as in the chlorophyll-deficient N. rustica mutant.
131. K. E. Drake, J. M. Moore, P. Bertrand, B. Fortnum, P. Peterson, and R. S. Lewis, “Black Shank Resistance and Agronomic Performance of Flue-Cured Tobacco Lines and Hybrids Carrying the Introgressed Nicotiana Rustica Region, Wz,” Crop Science, vol. 55, no. 1, pp. 79–86, 2015. doi: 10.2135/cropsci2014.02.0164. Abstract
     Black shank, caused by Phytophthora nicotianae, is one of the most important diseases affecting tobacco (Nicotiana tabacum L.) production in the United States. Genetic mechanisms are needed that provide resistance to current races and that can be combined into cultivars that provide high yields of cured leaf with acceptable quality. Previous research identified DNA markers associated with an introgressed N. rustica genomic region (designated as Wz) found to contribute to resistance to race 0 and race 1 isolates. Objectives of the current research were to use DNA markers to transfer Wz into the elite genetic background of flue-cured tobacco cultivar K 326 and to develop nearly isogenic lines and hybrids with and without the race 0 immunity gene Php. These materials were evaluated in multiple environments for black shank resistance, yield, and quality characteristics. Wz was observed to positively affect resistance in the seven diverse disease environments tested. Genotypes in which Wz was combined with Php exhibited the greatest levels of resistance. No evidence of a negative relationship between Wz and yield and/or quality was observed. Data suggest commercial value for Wz in flue-cured tobacco-breeding programs with the goal of developing high-yielding tobacco cultivars with resistance to race 0 and race 1. Further studies are necessary to determine the durability of Wz-mediated resistance, however.
132. M. Dudek, “The effect of varying doses of gamma radiation on morphological and cytogenetical characteristics in Nicotiana tabacum L. and Nicotiana rustica L.,” Biuletyn Instytutu Hodowli i Aklimatyzacji Roslin, no. No. 1/2, pp. 123–125, 1970. https://www.cabdirect.org/cabdirect/abstract/19721602040. Abstract
     Seed of 22 cultivars and three intervarietal hybrids was irradiated in 1962 and seed of four tobacco cultivars and one of N. rustica in 1965. Germination was reduced to varying extents in different forms, the most sensitive being Flandria and N. rustica ’Brazylijska’ [Brazilian], with a survival of 4% and 1% respectively after irradiation with 25 kR. Changes in leaf and flower colour and other...
133. S. Eapen, T. S. Rangan, M. S. Chadha, and M. R. Heble, “Morphogenetic and Biosynthetic Studies on Tissue Cultures of Atropa Belladonna L.,” Plant Science Letters, vol. 13, no. 1, pp. 83–89, Sep. 1978. doi: 10.1016/0304-4211(78)90068-8. Abstract
     The morphogenetic and biosynthetic ability of tissue cultures established from haploid and diploid plants of Atropa belladonna L. have been investigated. It was observed that the haploid tissue regenerated plantlets much more readily than did the diploids. The regenerants obtained in vitro were successfully transplanted to soil and grown to flowering stage. Haploid and diploid calli, regenerated young shoot buds and the in vitro obtained plants (at the stage of flowering) were subjected to chemical analysis. The total alkaloid contents in the calli and young shoot buds were very low (8–20 μg/g) as compared to the mature plant (1160 μg/g). Tropine, atropine, scopolamine and several unidentified Dragendorff-positive components were observed in the cultures. The alkaloid spectrum of the shoot buds was comparable to that of the unorganized calli. However, the relative proportion of atropine was more in the shoot buds as compared to the callus cultures. The regenerated flowering plants contained the principal belladonna alkaloids in quantities comparable to the plants raised from seeds.
134. E. M. East, “Genetic Reactions in Nicotiana. I. Compatibility,” Genetics, vol. 20, no. 5, pp. 403–413, Sep. 1935. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1208622/.
135. F. W. Eastwood, I. Kirson, D. Lavie, and A. Abraham, “New Withanolides from a Cross of a South African Chemotype by Chemotype II (Israel) in Withania Somnifera,” Phytochemistry, vol. 19, no. 7, pp. 1503–1507, Jan. 1980. doi: 10.1016/0031-9422(80)80205-6. Abstract
     Three new withanolides have been isolated from hybrids obtained by crossing a chemotype of Withania somnifera received from South Africa and chemotype II originating in Israel. The compounds have been characterized as 4β,20α-dihydroxy-1-oxo-5β,6β-epoxy-20R,22R-witha-24-enolide, 20α-hydroxy-1,4-dioxo-5β,6β-epoxy-20R,22R-witha-2,24-dienolide, and 20α-hydroxy-1,4-dioxo-5β,6β-epoxy-20R,22R-witha-2-enolide. The major steroid of the plant is withanolide D, while the other known withanolides present are 4β,20α-dihydroxy-1-oxo-5β,6β-epoxy-20R,22R,24S,25R-witha-2-enolide and withaferin A. The structures assigned to the new compounds are based on spectral evidence, analysis of their fragmentation under electron impact, and on chemical correlation with known compounds. The formation of these withanolides in this new hybrid is discussed briefly.
136. S. A. Eghis, “On the problem of the origin of Nicotiana rustica.,” Doklady Akademii nauk SSSR, vol. 26, pp. 952–56, 1940. https://www.cabdirect.org/cabdirect/abstract/19421600529. Abstract
     The varieties of N. rustica, are described. Mention is also made of the occurrence in Peru and neighbouring countries of a number of closely related species of the rustica group (these same species are placed in the paniculata group by Goodspeed). An amphidiploid, N. rustica synthetica (2n = 48). was obtained by colchicine treatment of the axillary buds of a hybrid between N. paniculata. (2n =...
137. E. G. Eglington and C. A. Moore, “The Nature of the Inheritance of Permanently Induced Changes in Nicotiana Rustica II. F4 and F5 Generations of Selected Crosses,” Heredity, vol. 30, no. 3, pp. 387–395, Jun. 1973. doi: 10.1038/hdy.1973.47. Abstract
     The F4 and F5 generations of the two selected crosses, nk2 × nil1 and p3 × nil3, from a dialle of all possible crosses between the three conditioned lines p, nil and nk were grown in successive years in the summers of 1970 and 1971 respectively in order to study (1) the persistence of the segregation between F1 individuals seen in earlier generations and (2) the conventional segregation in the later generations. In the F4 generation V1 segregation in the cross nk2 × nil1 approaches significance for the character final height. In the F6 generation significant segregation between F1 individuals was not found for the characters final height and flowering time for either cross. However, in the cross nk2 × nil1 significant F1 segregation was found for two other characters, height at flowering time and the diameter of the eighth leaf. The presence of segregation at the F2, F3 and F4 levels was confirmed for both characters in both crosses by the fitting of a standard additive genetical and additive environmental model. From the estimates of D obtained the numbers of effective factors by which these lines differ were found to range from three to five.
138. S. R. and R. Elango, “Effect of Microbial Consortium on Plant Growth and Improvement of Alkaloid Content in Withania Somnifera (Ashwagandha),” Current Botany, 2011. https://updatepublishing.com/journal/index.php/cb/article/view/1371. Abstract
     The effect of microbial consortium consisting Plant Growth Promoting Rhizobacteria (PGPR) like Azospirillum, Azotobacter, Pseudomonas and Bacillus were tested separately and in combination on Withania somnifera for two consecutive years (2009 and 2010). The combinations of above mentioned PGPR strains significantly increased plant height, root length, and alkaloid content in Withania somnifera when compared to the uninoculated control. Plant growth promoting rhizobacteria (PGPR) exhibit direct and indirect mechanisms as plant growth promoters and biological control agents. Direct mechanism by PGPR, include the provision of bio-available phosphorus for plant uptake, nitrogen fixation for plant. The results of this study suggest that the PGPR applied in combination have the potential to increase the plant growth, alkaloid content of Withania somnifera.
139. M. Elsakka, M. Pavelescu, and E. Grigorescu, “Withania Somnifera, a Plant with a Great Therapeutical Future,” Revista medico-chirurgicala a Societatii de Medici si Naturalisti din Iasi, vol. 93, no. 2, pp. 349–350, Apr. 1989. Abstract
     Withania somnifera Dun is a plant which drew the interest of many researchers in several countries, either for its active principles or for the extremely important pharmacodynamic or pharmacotherapeutic properties it has and first of all for the content in withanolides, substances with a sterol structure.
140. G. Elzenga, L. Smeets, and J. W. De Bruyn, “Influence of the Temperature on Growth and Alkaloid Content of First-Year Atropa Belladonna L.,” Euphytica, vol. 5, no. 3, pp. 276–280, Oct. 1956. doi: 10.1007/BF00038849. Abstract
     In breeding Atropa belladonna for high alkaloid content it is of importance to know whether the alkaloid content is affected by the environment. Therefore some experiments on the influence of temperature were carried out in the phytotron of our Institute. As the results were in agreement with each other only one experiment is discussed in which the influence of three constant temperatures, viz. 20°, 23° and 26°C, on growth rate, dry matter, and alkaloid content of yellow-flowered Atropa belladonna L. was studied.
141. G. Elzenga and J. W. de Bruyn, “Interrelation of Alkaloid Content and Stage of Development of 1- and 2-Year-Old Atropa Belladonna L.,” Euphytica, vol. 5, no. 3, pp. 259–266, Oct. 1956. doi: 10.1007/BF00038847. Abstract
     A study was made of the course of the alkaloid content in the aerial parts of both 1- and 2-year-old A. belladonna during growth. This content is not steady. One-year plants reach the maximum content during or just after flowering, when the plant bears a few green berries, while 2-year plants reach the maximum content in a very young stage. During flowering, however, the content drops to a low level to reach a peak again when the plant bears a good many green berries.
142. K. C. Engvild, “Pollen Irradiation and Possible Gene Transfer in Nicotiana Species,” Theoretical and Applied Genetics, vol. 69, no. 5, pp. 457–461, Mar. 1985. doi: 10.1007/BF00251084. Abstract
     Progeny from crosses of Nicotiana langsdorffii with gamma irradiated pollen of Nicotiana alata ‘Crimson Bedder’ showed skewed segregation in the F2 favoring the maternal parent. This is probably not gene transfer in a strict sense, rather just an extreme case of reduced transmission of irradiated chromosomes, leading to massive overrepresentation of maternal genes. Gene transfer or mutational loss may explain some anomalous F1 plants. Segregation in the F2 progeny showed the presence of several genes from the irradiated pollen. Crosses of Nicotiana sylvestris, N. plumbaginifolia N. paniculata, and Petunia parodii with irradiated pollen from N. alata and Petunia hybrida showed no evidence of gene transfer, nor did experiments with irradiated mentor pollen. This indicates that gene transfer with irradiated pollen between non-crossing species or between species giving sterile hybrids is probably a rare phenomenon.
143. A. K. Essa, “Variations in Alkaloidal Constituents of Plant Tissue Cultures. [Nicotiana Tabacum, Nicotiana Rustica, Datura Stramonium and Hyoscyamus Niger],” Jan. 1987. https://www.osti.gov/etdeweb/biblio/6292408. Abstract
     The accumulation of tobacco and Solanaceae-tropane alkaloids in calluses and suspensions of Nicotiana tabacum, Nicotiana rustica, Datura stramonium and Hyoscyamus niger was the main concern of this work. Nicotine, anabasine and anatabine had regularly been found in tobacco callus tissues grown for several passages. For the first time, N. tabacum suspensions were shown able to accumulate anabasine, anatabine, anatalline, myosmine and nicotelline whereas N. rustica calluses, N-methyl-, N,N-dimethyl- and N-methyl-N-nitroso anilines. The aim of these experiments was an attempt to affect the yields and types of alkaloids produced. The interrelationship between nicotine and protein contents in N. tabacum and N. rustica calluses was investigated. The possible role of urea and sodium propionate as precursors of nicotine in tobacco suspensions was checked by feeding the latter with carbon-14 radioactive substrates. The scope and quantum of the principle alkaloidal components present in the source N. tabacum plants and D. stramonium and H. niger seeds were investigated to stand as references against in vitro production. Identification of the products found was made by using gas chromatography-mass spectrometry (GC-MS) and/or MS.
144. M. Euler and I. T. Baldwin, “The Chemistry of Defense and Apparency in the Corollas of Nicotiana Attenuata,” Oecologia, vol. 107, no. 1, pp. 102–112, 1996. https://www.jstor.org/stable/4221312. Abstract
     The morphological and chemical characteristics of flowers which attract pollinators present a dilemma for plants; advertising may increase the "apparency" of plants to their predators and some pollinators are also predators. We explore how a self-compatible disturbance species, Nicotiana attenuata, copes with this potential dilemma by examining the changes in emission of chemicals from flowers in response to pollination and herbivory. We propose that chemical changes induced by herbivory and pollination reflect the function of the chemicals in the plant. The emission of a single compound, benzyl acetone (BA, 4-phenyl-2-butanone), by flowers increases dramatically (50×) in the evening, peaking just after dark – a pattern of emission characteristic of moth-pollinated flowers. Pools of BA were found only in the outer lip of the corolla where pollinators come in contact with the flower, and diurnal changes in the size of the corolla pool closely paralleled the amount emitted by intact flowers throughout the day, as determined by headspace sampling. Pollination dramatically decreases both the pools of BA in the corolla and its emission from flowers. Similarly, nicotine, a broadly biocidal defense metabolite and an induced defense in vegetative and reproductive tissues, is also found in the headspace of flowers and is principally localized in the basal parts of the corolla below the attachment of the filaments and the nectar reward. Moreover, the dynamics of the corolla pools of BA and nicotine throughout the day are consistent with their roles in advertisement and defense, respectively. The corolla pools of nicotine are stable throughout the day except during the period of peak BA production and emission when nicotine pools decrease significantly. The co-ordinated increase in BA emission and decline in nicotine pools are not inexorably linked, because herbivory or mechanical damage to corolla tissue rapidly increases corolla nicotine pools without affecting the increase in BA pools. Similarly, leaf damage results in a slower, systemic increase in corolla nicotine pools during reproductive growth but again does not affect BA pools. Excised flowers emitted BA in a manner similar to that of intact flowers, and excision of a majority of flowers from a plant did not alter the BA emission patterns of the remaining flowers. We conclude that although N. attenuata’s defensive and advertisement chemistries respond synchronously to some environmental stimuli, the flowers’ chemical responses to pollinators and herbivores are distinct and the differences reflect their ecological roles. We propose that the cost-benefit framework of the optimal defense and apparency theories can be fruitfully applied to the allocation of defense metabolites and floral volatiles that function in pollinator attraction, and that this framework can be readily tested by manipulating the patterns of the emissions of plants in the field.
145. H. J. Evans and M. E. Weeks, “The Influence of Nitrogen, Potassium, and Magnesium Salts on the Composition of Burley Tobacco,” Soil Science Society of America Journal, vol. 12, no. C, pp. 315–322, 1948. doi: 10.2136/sssaj1948.036159950012000C0075x.
146. N. Fatima, N. Ahmad, and M. Anis, “Enhanced in Vitro Regeneration and Change in Photosynthetic Pigments, Biomass and Proline Content in Withania Somnifera L. (Dunal) Induced by Copper and Zinc Ions,” Plant Physiology and Biochemistry, vol. 49, no. 12, pp. 1465–1471, Dec. 2011. doi: 10.1016/j.plaphy.2011.08.011. Abstract
     In the present study the effect of inorganic nutrients (CuSO4 & ZnSO4) on morphogenic and biochemical responses from nodal explants in Withania somnifera L. was investigated. Incorporation of either Copper sulphate (25–200 μM) or Zinc sulphate (50–500 μM) in the optimized Murashige and Skoog (MS) medium highly influenced the shoot bud formation and subsequent elongation, which induced maximum percentage (95%) regeneration, number (61.7 ± 0.25) of shoots with shoot length (5.46 ± 0.16 cm) on CuSO4 (100 μM) and maximum percentage regeneration (100%), number of shoots (66.1 ± 0.96) with shoot length (6.24 ± 0.21 cm) on ZnSO4 (300 μM) after 12 weeks of culture. Healthy growing in vitro microshoots rooted efficiently on ½ MS medium supplemented with NAA (0.5 μM), which induced (16.2 ± 0.12) roots with root length (3.30 ± 0.12 cm) after 4 weeks. Pigment content increased with increasing concentration of Cu and Zn and the maximum Chl. a (0.47), (0.41); Chl. b (0.52), (0.42); total Chl. (0.99), (0.83) and Carotenoid (0.16), (0.16) mg/g FW contents in regenerants were found on CuSO4 (100 μM) and ZnSO4 (300 μM), respectively. Maximum proline content (0.17), (0.16) μg/g FW was observed on high concentrations of CuSO4 (200 μM) and ZnSO4 (500 μM) respectively, in the basal medium. Regenerated plantlets were acclimatized successfully in soilrite with a survival rate of 95%. No morphological variations were detected among the micropropagated plants when compared with seedling raised plants of the same age.
147. N. Fatima, N. Ahmad, M. Anis, and I. Ahmad, “An Improved in Vitro Encapsulation Protocol, Biochemical Analysis and Genetic Integrity Using DNA Based Molecular Markers in Regenerated Plants of Withania Somnifera L,” Industrial Crops and Products, vol. 50, pp. 468–477, Oct. 2013. doi: 10.1016/j.indcrop.2013.08.011. Abstract
     Non-embrogenic, synthetic seeds were produced by encapsulating nodal segments (containing axillary buds) of Withania somnifera L. in calcium alginate hydrogel containing Murashige and Skoog (MS) medium. A 3% sodium-alginate with 100mM calcium cloride found to be the optimum concentration for the production of uniform synthetic seeds. Effect of different treatments (M1–M5), i.e. MS medium containing different concentrations of cytokinins (0.5, 1.0, 2.5, 5.0 & 10.0μM) along with optimal level of auxins NAA (0.5μM) on in vitro morphogenic response of synthetic seeds was evaluated. The maximum frequency (86.2%) of conversion of encapsulated beads into plantlets was achieved on MS (M3) medium containing 6-benzyladenine, BA (2.5μM) and α-naphthalene acetic acid, NAA (0.5μM) after 4 weeks of culture. Rooting in plantlets was achieved on 1/2MS+NAA (0.5μM). Plantlets obtained from stored synthetic seeds were hardened, acclimatized and established in field, where they grew well without any detectable malformation. Significant enhancement in the pigment contents (chlorophyll, carotenoids and net photosynthetic rates) with an increase in acclimatization days may be attributed to chlorophyll biosynthesis. Activities of antioxidant enzymes i.e. superoxide dismutase, catalase and peroxidase activity) were significantly increased which suggests their preventive role in membrane oxidation and damage to biological molecules. Also, an enhanced level of lipid peroxidation, as indicated by MDA content, a sensitive diagnostic index of oxidative injury clearly indicating its positive determining role in combating oxidative stress during acclimatization of plantlets. The generated RAPD and ISSR profiles from regenerated plantlets with mother plant were monomorphic which confirms the genetic stability among the clones. This synthetic seed technology could possibly paves the way for the conservation, short-term storage, germplasm exchange with potential storability and limited quarantine restrictions.
148. N. Fatima, N. Ahmad, I. Ahmad, and M. Anis, “Interactive Effects of Growth Regulators, Carbon Sources, pH on Plant Regeneration and Assessment of Genetic Fidelity Using Single Primer Amplification Reaction (SPARS) Techniques in Withania Somnifera L.,” Applied Biochemistry and Biotechnology, vol. 177, no. 1, pp. 118–136, Sep. 2015. doi: 10.1007/s12010-015-1732-x. Abstract
     An improved and methodical in vitro shoot morphogenic approach through axillary bud multiplication was established in a drug yielding plant, Withania somnifera L. Effects of plant growth regulators [6-benzyladenine (BA), kinetin (Kin), 2-isopentenyladenine (2iP), and thidiazuron (TDZ)] either singly or in combination with α-napthalene acetic acid (NAA), indole-3-butyric acid (IBA), and indole-3-acetic acid (IAA) in Murashige and Skoog (MS) medium were tested. The highest regeneration frequency (90 %) with optimum number of shoots (32\,± 0.00)/explant were obtained on MS medium fortified with 2.5 μM 6-benzyladenine (BA) and 0.5 μM NAA and 30 g/l sucrose at pH 5.8. Among the tried TDZ concentrations, 0.5 μM resulted in maximum number of shoots (20.4\,± 0.40)/explant after 4 weeks of exposure. The proliferating shoot cultures established by repeated subculturing of the mother explants on the hormone-free medium produced the highest shoot number (29.4\,± 0.40) with shoot length (6.80\,± 0.12 cm)/explant at fourth subculture passage, which a decline in shoot proliferation was recorded. Different concentrations of NAA were tested for ex vitro rooting of microshoots. The maximum percentage of rooting 100 % with maximum roots (18.3\,± 0.1) was achieved in soilrite when basal portion of the microshoots were treated with 200 μM (NAA) for 15 min per shoot. The plantlets went through hardening phase in a growth chamber, prior to ex vitro transfer. The PCR-based single primer amplification reaction (SPAR) methods which include random amplified polymorphic DNA (RAPD) and direct amplification of minisatellite DNA (DAMD) markers has been used for assessment of genetic stability of micropropagated plantlets. No variation was observed in DNA fingerprinting patterns among the micropropagated and the donor plants illustrating their genetic uniformity.
149. I. D. N. S. Fernando, D. C. Abeysinghe, and R. M. Dharmadasa, “Determination of Phenolic Contents and Antioxidant Capacity of Different Parts of Withania Somnifera (L.) Dunal. from Three Different Growth Stages,” Industrial Crops and Products, vol. 50, pp. 537–539, Oct. 2013. doi: 10.1016/j.indcrop.2013.08.042. Abstract
     Withania somnifera (L.) Dunal. (Solanaceae) is a therapeutically important medicinal herb used in Ayurvedic and traditional systems of medicine for the treatment of an array of ailments. Diverse therapeutic properties reported from W. somnifera are mainly due to the high content of polyphenols and antioxidant activities present in different parts of the plant. Present study investigates the total phenolic content (TPC) and total antioxidant capacity (TAC) of different parts of three different growth stages of W. somnifera grown under three different spacing levels. The TAC and TPC were determined using Ferric Reducing Antioxidant Power assay and modified Folin–Ciocalteu colorimetric method respectively. Leaf extract exhibited significantly higher (p<0.05) TAC and TPC values for all three different growth stages. However, the highest TAC and TPC of leaf extract for all three spacing levels were observed just after flowering stage. The highest total phenolic content was exhibited in leaf extracts followed by flower, fruits, stem and roots respectively. With regard to the anti oxidant content, the highest amount was recorded from leaf followed by pods, flowers, stem and roots respectively. Presence of higher TPC and TAC just after flowering stage scientifically validates traditional claims of harvesting of W. somnifera after flowering stage. The higher content of TPC and TAC in leaf demonstrated the possibility of incorporation leaf for the development of newer, effective drugs instead of roots.
150. S. K. Florentine, M. E. Westbrooke, K. Gosney, G. Ambrose, and M. O’Keefe, “The Arid Land Invasive Weed Nicotiana Glauca R. Graham (Solanaceae): Population and Soil Seed Bank Dynamics, Seed Germination Patterns and Seedling Response to Flood and Drought,” Journal of Arid Environments, vol. 66, no. 2, pp. 218–230, Jul. 2006. doi: 10.1016/j.jaridenv.2005.10.017. Abstract
     Disturbances in plant communities provide opportunities for weed germination, propagation, spread, and invasion. When the population density of a weed increases, fast-tracked and appropriate control management strategies are required. The objectives of this study were to: (i) examine the population and soil seed bank dynamics of Nicotiana glauca; (ii) compare the germination patterns of invasive N. glauca seeds collected from two states in Australia, and (iii) investigate the impact of a flood in September 1997 and subsequent drought on N. glauca seedlings. The density of N. glauca followed a steep positive increment during the sampling time (September 1999 to October 2004). The increment pattern was similar in flooded fenced and unfenced plots. Plant density increased over much of the observation period, but had declined to 80 and 432 stemsha−1, respectively, by October 2004. Stem density recorded in October 2004 along two transects radiating from the central point of the newly created lake demonstrated that a significant number of stems appeared to be dead. A soil seed bank study revealed that seed density varied significantly (p=0.0001) between flooded fenced (598.75±71) and flooded unfenced (327.5±66) plots. In contrast, no N. glauca seedlings were recruited from the soil collected from the control plots. Germination trials were undertaken on N. glauca seed collected from New South Wales. There was no significance difference detected between treatments light and temperature. Similarly, no interaction was found between light and temperature. A comparative study on seed germination patterns of N. glauca seeds collected from Ivanhoe, New South Wales, and the Flinders Ranges, South Australia, showed that temperature had a significant effect on N. glauca seed germination. The effect varied significantly with main variables (state, length and time) and also with different interactions, except state×light (p=0.3840). N. glauca seedlings exposed to flood were found to withstand partial flooding for at least 58 days. Under waterlogged conditions, the seedlings showed stem hypertrophy and produced adventitious roots. Only one seedling was found dead in the drought treatment. In conclusion, it is clear that N. glauca invaded the area after a rare flood event and began to function as a casual weed. Established seedlings in the field can withstand extreme ecological events such as flood and drought. Understanding the plants’ ecological characteristics through a study such as this at an early rather than late stage in the invasion will help us to take appropriate control measures for this species.
151. S. K. Florentine et al., “Influence of Selected Environmental Factors on Seed Germination and Seedling Survival of the Arid Zone Invasive Species Tobacco Bush (Nicotiana Glauca R. Graham),” The Rangeland Journal, vol. 38, no. 4, pp. 417–425, Jul. 2016. doi: 10.1071/RJ16022. Abstract
     Tobacco bush (Nicotiana glauca R. Graham) is an aggressive invading species, which is active after disturbances such as high rainfall events and flooding. Past studies have focussed on population dynamics and allelopathic effects associated with the species, but little is known about its seed ecology. To address this dearth of information, this study aimed to investigate: (i) the effect of various stress factors (temperature, light, osmotic potential, salt stress, heat-shock, a combination of heat-shock and smoke, pH buffer, and burial depth of seed) on seed germination and seedling emergence, and (ii) factors affecting the fate of seedlings. The results show that N. glauca was able to germinate over a broad range of temperatures with highest seed germination occurring at 30/20°C with 12 h of light and 12 h of dark conditions. Seed germination was greatest (89%) when seeds were placed on the soil surface and emergence decreased considerably as planting depth increased from 0.5 to 1.5 cm. Water stress greatly reduced seed germination (58% germination at osmotic potentials below –0.2 MPa) and germination was completely inhibited at water potentials of –0.4 to –0.6 MPa. Although increasing salinity reduced the seed germination of this invasive species, N. glauca seed was able to germinate in both alkaline (81% at pH 10) and acidic (80% at pH 4) conditions. The trial on the effect of seed age and field seed burial on seed germination showed a slight decline in seed germination after 120 days of burial compared with non-buried seeds. Further, the combined effect of heat-shock and smoke effectively inhibited the germination of N. glauca seeds; however, our study shows that seedlings of N. glauca can withstand heat-shock of up to 130°C. Studies such as this will assist in the development of control strategies to prevent the spread of this invasive species into arid landscapes.
152. Y. Fracheboud and P. J. King, “An Auxin-Auxotrophic Mutant of Nicotiana Plumbaginifolia,” Molecular and General Genetics MGG, vol. 227, no. 3, pp. 397–400, Jul. 1991. doi: 10.1007/BF00273929. Abstract
     Auxin (indole-3-acetic acid) is considered to be an important signalling molecule in the regulation of plant growth and development but neither auxin synthesis nor its mode of action is clearly understood. To identify genes involved in these processes, mutations were sought that altered the auxin requirement of plant tissues for growth. For the first time mutant plants were obtained that carry a recessive mutation at a single nuclear locus (auxl) which results in an absolute requirement for exogenous auxin for normal growth. In the absence of auxin treatment, mutant plants undergo premature senescence and die.
153. V. Fragoso, H. Goddard, I. T. Baldwin, and S.-G. Kim, “A Simple and Efficient Micrografting Method for Stably Transformed Nicotiana Attenuata Plants to Examine Shoot-Root Signaling,” Plant Methods, vol. 7, no. 1, p. 34, Oct. 2011. doi: 10.1186/1746-4811-7-34. Abstract
     To adjust their development to the environment, plants rely on specific signals that travel from shoot to root and vice versa. Here we describe an efficient micrografting protocol for Nicotiana attenuata, a useful tool for identifying these signals and understanding their functions. Additionally we analyzed transcript accumulation profiles of scions and rootstocks of grafts performed with wild-type and stably transformed N. attenuata. Our results are consistent with the source-to-sink movement of an sRNA silencing signal.
154. G. M. Frost, K. S. Yang, and G. R. Waller, “Nicotinamide Adenine Dinucleotide as a Precursor of Nicotine in Nicotiana Rustica L,” Journal of Biological Chemistry, vol. 242, no. 5, pp. 887–888, Mar. 1967. doi: 10.1016/S0021-9258(18)96207-1. Abstract

     Quinolinic acid-2,3,7,8-¹⁴C, nicotinic acid-2,3,7-¹⁴C, and nicotinamide adenine dinucleotide-2,3,7-¹⁴C were shown to be incorporated into nicotine by Nicotiana rustica L. with very similar efficiencies. It is postulated that compounds of the pyridine nucleotide cycle, previously implicated as a source of metabolic intermediates for the biosynthesis of ricinine in Ricinus communis L., could serve the same function in nicotine biosynthesis in the tobacco plant.

155. J. M. Furze, J. D. Hamill, A. J. Parr, R. J. Robins, and M. J. C. Rhodes, “Variations in Morphology and Nicotine Alkaloid Accumulation in Protoplast-Derived Hairy Root Cultures of Nicotiana Rustica,” Journal of Plant Physiology, vol. 131, no. 3, pp. 237–246, Dec. 1987. doi: 10.1016/S0176-1617(87)80163-3. Abstract
     A suspension culture was produced by the phytohormone treatment of hairy roots resulting from infection of Nicotiana rustica with Agrobacterium rhizogenes. This was used as a source of protoplasts for the production of single-cell-derived clones. After conditions for protoplast release were optimised, it was possible to induce high rates of cell division from these protoplasts and regenerate hairy roots from the protoplast-derived calli following return to hormone-free medium. Regenerated roots were assessed for nicotine alkaloid accumulation and their transformed nature was confirmed by DNA analysis. Variation between clones was observed in alkaloid synthesis, morphology and T-DNA structure, with some of the clones showing improved alkaloid synthesis characteristics relative to the parent line.
156. M. Ganzera, M. I. Choudhary, and I. A. Khan, “Quantitative HPLC Analysis of Withanolides in Withania Somnifera,” Fitoterapia, vol. 74, no. 1, pp. 68–76, Feb. 2003. doi: 10.1016/S0367-326X(02)00325-8. Abstract
     One of the most widely used herbs in Ayurvedic medicine is Ashwaghanda, Withania somnifera, a shrub commonly found on the Indian subcontinent. As this plant is increasingly becoming a popular adaptogenic in the western world, analytical methods for its identification and quality control are in demand. Thus, a HPLC method for the determination of withaferin A and withanolide D was developed. The system was successfully used to investigate the presence of the markers in different W. somnifera plant parts as well as to analyze their content in market products.
157. S. K. Gaur and K. Kumar, “A Comparative Bioefficacy of Seed and Root Extracts of a Medicinal Plant, Withania Somnifera When Administered to Prepupae of Lepidopteran Insects, Spodoptera Litura (Lepidoptera: Noctuidae) and Pericallia Ricini (Lepidoptera: Arctiidae),” The Journal of Basic and Applied Zoology, vol. 80, no. 1, p. 37, May 2019. doi: 10.1186/s41936-019-0107-1. Abstract
     The lepidopterans, Spodoptera litura and Pericallia ricini, are polyphagous pests of agricultural importance. The unscrupulous and non-judicious use of chemical pesticides for controlling the insect pests has resulted into severe environmental hazards, threatening non-target organisms and human health.
158. R. Gaur and K. Kumar, “Insect Growth-Regulating Effects of Withania Somnifera in a Polyphagous Pest, Spodoptera Litura,” Phytoparasitica, vol. 38, no. 3, pp. 237–241, Jul. 2010. doi: 10.1007/s12600-010-0092-x. Abstract
     Treatment of sixth instar larvae and pupae of the polyphagous pest Spodoptera litura Fabr. (Lepidoptera: Noctuidae) with an acetone extract of leaves of Ashwagandha, Withania somnifera L. (Solanaceae), caused toxicity, molt disturbances, formation of larval–pupal, pupal–adult intermediates and adultoids. Our results suggest that W. somnifera acts as an insect growth regulator causing disruption of the endocrine mechanism regulating molting and metamorphosis.
159. S. K. Gaur and K. Kumar, “Sensitivity of Tobacco Caterpillar, Spodoptera Litura, to Extract from a Medicinal Plant, Withania Somnifera,” International Journal of Vegetable Science, vol. 26, no. 1, pp. 62–78, Jan. 2020. doi: 10.1080/19315260.2019.1605556. Abstract
     The larval stage of a polyphagous moth Spodoptera litura (Lepidoptera: Noctuidae), commonly known as tobacco caterpillar, causes loss in many vegetables. Use of chemical pesticides to control this pest has resulted in environmental pollution, is hazardous to non-target organisms, and contributes to development of pest resistance against these compounds. Alternatives for pest management need to be developed. Extracts derived from plants can function as insect growth regulators and are easily biodegradable. Topical administration of root extracts from a medicinal plant, Ashwagandha (Withania somnifera L. Dunal), to sixth instar larvae and freshly molted pupae (0–2 h old) of S. litura caused prolongation of larval-pupal and pupal-adult ecdysis duration, mortality, reduced pupation, and adult emergence. Root extracts of W. somnifera produced the morphological and developmental abnormalities like ecdysial failure, formation of larval-pupal and pupal-adult intermediates, abnormal pupae and adultoids. Treatment with root extracts of W. somnifera was more toxic to pupae compared to larvae of S. litura. The results may be due to interference with normal hormonal mechanism eventually leading to disruption of molting and metamorphosis.
160. S. K. Gaur and K. Kumar, “Toxicity and Insect Growth Regulatory Activities of Medicinal Plant, Withania Somnifera, in Flesh Fly, Sarcophaga Ruficornis (Diptera: Sarcophagidae),” The Journal of Basic and Applied Zoology, vol. 81, no. 1, p. 30, May 2020. doi: 10.1186/s41936-020-00154-2. Abstract
     The flesh fly Sarcophaga ruficornis is well known for its medical and veterinary importance in causing myiasis in humans and animals. The conventional use of chemical pesticides for controlling insect pests has resulted in environmental pollution besides posing serious hazards to non-target organisms and the development of pest resistance against these compounds. Considering the various harmful and adverse effects of chemical pesticides, an alternative and safe approach for the pest management has to be explored. The plant extracts derived from plants are eco-friendly in nature, easily biodegradable, and can be used as botanical pesticides.
161. S. K. Gaur and K. Kumar, “Toxicity and Insect Growth Regulatory Effects of Root Extract from the Medicinal Plant, Withania Somnifera (Linnaeus) against Red Flour Beetle, Tribolium Castaneum (Coleoptera: Tenebrionidae),” Archives of Phytopathology and Plant Protection, vol. 53, no. 17-18, pp. 856–875, Oct. 2020. doi: 10.1080/03235408.2020.1802566. Abstract
     The insect growth regulatory effects of the medicinal plant, Withania somnifera (Family: Solanaceae) on the moulting and metamorphosis of economically important stored grain pest, Tribolium castaneum (Coleoptera: Tenebrionidae) were assessed under laboratory conditions. Seventh instar larvae of the red flour beetle, T. castaneum were topically treated with different doses of root extract of W. somnifera. Treatment with root extract of W. somnifera disrupted growth, development, moulting and metamorphosis in T. castaneum. The results included mortality, prolongation of larval-pupal and pupal-adult ecdysis, ecdysial stasis, development of larval-pupal and pupal-adult intermediates and adultoids, eclosion inhibition, reduced pupation and adult emergence. The LC50 value in topical application of root extract of W. somnifera was 0.5850 µg·µL−1 against last instar larvae of T. castaneum. These results are similar to those observed by the application of juvenile hormone analogues and may be due to interference with the normal hormonal mechanism of moulting and metamorphosis.
162. S. K. Gaur and K. Kumar, “Withania Somnifera Acts as a Potential Insect Growth Regulator in the Polyphagous Pest, Pericallia Ricini,” Journal of Plant Protection Research, vol. 57, no. 4, 2017. doi: 10.1515/jppr-2017-0052.
163. N. Gaurav and A. Kumar, “Effect of Growth Regulators on In-Vitro Callusing of Wild Variety of Withania Somnifera L. in B5 Medium.,” Indian Forester, vol. 145, no. 12, pp. 1176–1181, 2019. https://www.cabdirect.org/cabdirect/abstract/20203122766. Abstract
     "Ashwagandha" products are one of the major herbal components of various disease tonics mentioned in Indian systems of medicine. It is an erect branching under shrub reaching about 150 cm in height, usually clothed with minutely stellate tomentum; leaves ovate up to 10 cm long, densely hairy beneath and sparsely above, flowers are greenish or yellow in axillary fascicles, bisexual, pedicel long,...
164. E. Genova, G. Komitska, and Y. Beeva, “Study on the Germination of Atropa Bella-Donna L. Seeds,” Bulgarian Journal of Plant Physiology, vol. 23, no. 1-2, pp. 61–66, 1997. http://www.bio21.bas.bg/ipp/gapbfiles/v-23/97_1-2_61-66.pdf. Abstract
     The germination of Atropa bella-donna L. seeds, a medicinal species with restrictive mode of use, was studied. It was established that variable temperature (6 h at 30 °C and 18 h at 15 °C) significantly stimulates seed germination – 82.5%. A maximum germination was obtained by treatment with gibberellic acid (GA3) 1mg/l H2O – 89.5%
165. T. K. George, J. Houbraken, L. Mathew, and M. S. Jisha, “Penicillium Setosum, a New Species from Withania Somnifera (L.) Dunal,” Mycology, vol. 10, no. 1, pp. 49–60, Jan. 2019. doi: 10.1080/21501203.2018.1555868. Abstract
     Medicinal plants are considered as sources of novel and unexplored groups of endophytic microorganisms. A study on endophytic fungal species from the medicinal plant Withania somnifera (L.) Dunal resulted in the isolation of a Penicillium isolate (WSR 62) with antibiotic activity. Phylogenetic analysis showed that the isolate belongs to section Lanata-divaricata, and it is most closely related to P. javanicum. Subsequent detailed phylogenetic analyses using partial β-tubulin (BenA), calmodulin (CaM) and DNA-dependent RNA polymerase II (RPB2) gene sequences of a larger number of related strains revealed the distinctiveness of the isolate in the P. javanicum-clade. The isolate grows fast on Czapek yeast autolysate agar (CYA) and malt extract agar (MEA) incubated at 25°C, 30°C and 37°C. The obverse colony colour is dominated by the conspicuous production of cleistothecia and is greyish yellow on CYA and yellowish brown on MEA. Production of cleistothecia containing prominent spinose ascospores was present on all tested agar media. Based on the phylogenetic analysis and the phenotypic characterisation, strain WSR 62 from Withania is described here as a novel species named Penicillium setosum.
166. H. Ghader, S. Zienab, and H. Roghieh, “Effect of Salicylic Acid on Tobacco (Nicotiana Rustica) Plant Under Drought Conditions,” vol. 7, no. 25, pp. 17–28, Jan. 2015. https://www.sid.ir/paper/159996/fa. Abstract

     Drought stress impact photosynthesis and stomatal conductance, and may reduce the overall production capacity of plants. Since exogenous application of salicylic acid (SA) can partially alleviatebe increased the negative effects tolerance of drought stress by improve the metabolism pathways and increase the net photosynthesison plant photosynthesis and metabolism, the main objective of this study was to clarify the roles of SA in enhancing 28 days tobacco (Nicotiana rustica Basmas) tolerance to drought stress (50% FC). The results indicated that foliar application of SA (0.5 mM) influenced negatively net CO\textsubscript2 assimilation rate and stomatal conductance and led to reduction of shoot and root dry masses. In contrast, the stress did not reduce significantly the maximal quantum yield of photosystem II (PSII). This that can be explained by enhancement of efficiency for dissipation of excess photon energy in the PSII antenna, determined as non-photochemical quenching, and consequently further protection of PSII from photodamage. Thus, under more drought stress, the reduction of photosynthesis of tobacco plants was due mainly to reduction of stomatal conductance. Under water-deficient conditions, plants showed an increase in chlorophyll a and amino acids concentrations in the leaves when treated with SA while this change for net photosynthesis was negligible. Our results indicated showed that the foliar application of SA had no ameliorative effect on tobacco growth under drought stress, because its effect on elevation of transpiration rate did not increase net photosynthesis under drought condition.

167. B. Ghimire and K. Heo, “Embryology of Withania Somnifera (L.) Dunal (Solanaceae),” Acta Biologica Cracoviensia. Series Botanica, vol. 54, no. 2, 2012. doi: 10.2478/v10182-012-0027-6.
168. B. Ghimire, B. K. Ghimire, and K. Heo, “Seed Characteristics of Withania Somnifera (Solanaceae),” Korean Journal of Plant Taxonomy, vol. 41, no. 2, pp. 103–107, 2011. doi: 10.11110/kjpt.2011.41.2.103. Abstract
     가지과 Withania somnifera (L.) Dunal의 종자 해부학적 특징을 알기 위하여 광학현미경과 주사전자 현미경을 사용하여 관찰하였다. 종자의 색깔은 노랑색이고, 종자의 형태는 신장형이며, 크기가 2−3 mm 정도이다. 종자는 종의를 가지고 있지 않았으며, 배유가 잘 발달한 유배유종자였다. 종피형태는 외종피외층형으로 나타났다. 종피는 외종피만으로 구성되는 단주피성 이었으며, 한 층의 외종피외층, 여러 층의 외종피중층, 그리고 한 층의 외종피내층으로 구성되었다. 그러나 종자가 완전히 성숙했을 때는 외종피중층이 모두 압착되었으며, 외종피외층은 특징적으로 파상형의 후벽세포로 발달하였다. 종피의 표면은 망상구조를 가지며, 수층 벽구조는 파상형으로 발달하였고, 표면에 여러 개의 구멍이 발달하는 특징이 관찰되었다. The seed characteristics of Withania somnifera were studied using light and scanning electron microscopy in order to determine the specific features of this species. The seed color is yellow, and the seed shape is reniform measuring between two to three millimeters. The seed of W. somnifera is exarillate and albuminous. The seed coat type is exotestal. The seed coat develops from a single integument. The young seed coat consists of single-layered exotesta, multi-layered mesotesta and single-layered endotesta. However, parenchymatous mesotesta layers are completely compressed at maturity. Therefore, the seed coat was represented by sclerenchymatous exotesta. The primary sculpture on the seed surface is reticulate, and cells are irregular in shape with undulating anticlinal walls. In addition, the seed surface has several characteristic holes between the anticlinal walls.
169. S. Ghoshal, C. Ghule, A. Mirgal, A. Girme, and L. Hingorani, “Recent Findings by High-Performance Thin-Layer Chromatographic Separation for a Comprehensive Analysis of Withania Somnifera by Densitometry and Mass Spectrometry: An Assessment to Quality and Adulteration,” JPC – Journal of Planar Chromatography – Modern TLC, vol. 35, no. 5, pp. 439–451, Nov. 2022. doi: 10.1007/s00764-022-00187-z. Abstract
     Withania somnifera (L.) Dunal (WS), also known as Ashwagandha, is an Indian herb classified as Rasayana in Ayurveda owing to its rejuvenating and health-promoting activities. A sensitive and robust high-performance thin-layer chromatography (HPTLC) method was developed and validated for the estimation of withanoside IV (WS4), withanoside V (WS5), withaferin A (WFA), and kaempferol-based glucoside (KRG) in the roots and aerial parts of WS. The present rapid HPTLC method reports the separation and simultaneous quantification of three diverse classes of WS as withanolide (WFA), withanosides (WS4 and WS5) with an analytical marker flavonoid glycoside (KRG) for effective detection, of the aerial parts separating them from roots. In normal-phase TLC plates, these markers showed good resolution in ethyl acetate‒chloroform‒methanol‒water (8:3:4.4:1.8, V/V) solvent system. Densitometric analysis was performed for KRG (at 254 nm), WFA, WS4, and WS5 (at 540 nm after derivatization) with characterization by high-performance thin-layer chromatography‒tandem mass spectrometry (HPTLC‒MS/MS). The method was found linear (R2\,> 0.99) for KRG and WFA (200‒1000 ng/band) and WS4‒WS5 (400‒2000 ng/band) with excellent recoveries (80‒100%) for all compounds. In this HPTLC based assessment, three compounds of WS could be rapidly detected for quality control distinguishing from other Withania and plant species reported as adulterants. Additionally, this method separated pigment-based compounds at 366 nm in chemical fingerprinting in WS aerial parts samples. Therefore, the presence of KRG as an analytical marker with these pigment zones can identify aerial parts, leaves, stems, fruits with calyx, and fruits in the HPTLC analysis, separating them from root samples. Hence, this HPLTC method was found robust and economical for the estimation and surety of the quality of WS samples in quantitative and qualitative analysis.
170. R. Gill, A. Rashid, and S. C. Maheshwari, “Isolation of Mesophyll Protoplasts of Nicotiana Rustica and Their Regeneration into Plants Flowering in Vitro,” Physiologia Plantarum, vol. 47, no. 1, pp. 7–10, 1979. doi: 10.1111/j.1399-3054.1979.tb06502.x. Abstract
     Protoplasts were isolated from leaves of in vitro grown plants of Nicotiana rustica L. by the one step enzymatic method. With 3% cetlulase in 0.5 M mannitol at 25°C and pH 4.6, within 10–12 h about half the total cells were transformed into protoplasts. The enzyme activity had two pH maxima, one at pH 3.5 and the other at pH 6.5, indicating the presence of isoenzymes. A time-course study at different temperatures indicated that ai 30°C the protoplasts’ liberation was quicker but it resulted in their subsequent bursting. The protoplasts were cultured in liquid as well as on agar-jelled medium of Ohyama and Nitsch (1972) supplemented with 2.4-D and benzylaminopurine, each at I mg/1. and 14% sucrose. Regular divisions could be seen only on agar-jelted medium. Colonies on transfer to Murashige and Skoog (1962) medium, containing IAA and kinetin, differentiated into plantlets. These plants, within 2 months, flowered in vitro and set seed of which about 20% germinated.
171. A. P. Giri et al., “Molecular Interactions between the Specialist Herbivore Manduca Sexta (Lepidoptera, Sphingidae) and Its Natural Host Nicotiana Attenuata. VII. Changes in the Plant’s Proteome,” Plant Physiology, vol. 142, no. 4, pp. 1621–1641, Dec. 2006. doi: 10.1104/pp.106.088781. Abstract
     When Manduca sexta attacks Nicotiana attenuata, fatty acid-amino acid conjugates (FACs) in the larvae’s oral secretions (OS) are introduced into feeding wounds. These FACs trigger a transcriptional response that is similar to the response induced by insect damage. Using two-dimensional gel electrophoresis, matrix-assisted laser desorption ionization-time of flight, and liquid chromatography-tandem mass spectrometry, we characterized the proteins in phenolic extracts and in a nuclear fraction of leaves elicited by larval attack, and/or in leaves wounded and treated with OS, FAC-free OS, and synthetic FACs. Phenolic extracts yielded approximately 600 protein spots, many of which were altered by elicitation, whereas nuclear protein fractions yielded approximately 100 spots, most of which were unchanged by elicitation. Reproducible elicitor-induced changes in 90 spots were characterized. In general, proteins that increased were involved in primary metabolism, defense, and transcriptional and translational regulation; those that decreased were involved in photosynthesis. Like the transcriptional defense responses, proteomic changes were strongly elicited by the FACs in OS. A semiquantitative reverse transcription-PCR approach based on peptide sequences was used to compare transcript and protein accumulation patterns for 17 candidate proteins. In six cases the patterns of elicited transcript accumulation were consistent with those of elicited protein accumulation. Functional analysis of one of the identified proteins involved in photosynthesis, RuBPCase activase, was accomplished by virus-induced gene silencing. Plants with decreased levels of RuBPCase activase protein had reduced photosynthetic rates and RuBPCase activity, and less biomass, responses consistent with those of herbivore-attacked plants. We conclude that the response of the plant’s proteome to herbivore elicitation is complex, and integrated transcriptome-proteome-metabolome analysis is required to fully understand this ubiquitous ecological interaction.
172. N. Gk, P. R, and G. G, “Integrated Nutrient Management in Ashwagandha (<Em>Withania Somnifera</Em>): A Review,” Journal of Pharmacognosy and Phytochemistry, vol. 7, no. 3S, pp. 118–121, 2018. https://www.phytojournal.com/special-issue/2018.v7.i3S.4737/integrated-nutrient-management-in-ashwagandha-ltemgtwithania-somniferaltemgt-a-review. Abstract
     <em>Withania somnifera</em> commonly called as Ashwagandha is a widely used medicinal herb in Ayurveda. It is considered to be a Rasayana herb, an adaptogen, and is commonly referred to as ‘Indian ginseng’. It is one of the most important medicinal plant coined with two words <em>viz</em>., Ashwini and Gandha. Ashwini means horse and Gandha means power. Although an important medicinal plant, it is still seen growing on waste and marginal lands, with little or no manures and fertilizers. Use of organic manures, biofertilizers and inorganic fertilizers has assumed great importance for sustainable production and for maintaining soil health. These not only supply macro- and micro-nutrients to the crop but also improve the physical, chemical and biological properties of the soil, leading to good crop production. The advantage of combining inorganic and organic sources of nutrients generally results in better use of each component which is responsible for crop growth and development. In this paper, the literatures pertaining to the different aspects of integrated nutrient management are reviewed.
173. S. Gleddie, W. A. Keller, G. Setterfield, and L. R. Wetter, “Somatic Hybridization between Nicotiana Rustica and N. Sylvestris,” Plant Cell, Tissue and Organ Culture, vol. 2, no. 4, pp. 269–283, Dec. 1983. doi: 10.1007/BF00039874. Abstract
     Protoplasts isolated from cell cultures of chlorophyll-deficient Nicotiana rustica cv. chlorotica and wild-type N. sylvestris were fused. The scheme for selection of somatic hybrids was based on the inability of the protoplast-derived colonies of the parental species to turn green; N. sylvestris protoplasts also had a very low plating efficiency in the medium employed. A total of 777 green colonies which were presumptive hybrids were isolated within four weeks of the fusion experiments. One hundred and eight green colonies formed shoots in vitro and 16 lines were rooted and grown in the greenhouse. Each of these hybrid plants displayed vegetative and floral traits intermediate to those of the parental species, except for plant height which in almost all cases was greater in the hybrids. Isozyme analyses by gel electrophoresis and isoelectric focussing of the small subunit of ribulose 1,5-bisphosphate carboxylase/oxygenase (RUBPCase) demonstrated that the nuclear genomes of both parents were expressed by the hybrids. Each of the eight somatic hybrid plants analyzed expressed only the N. rustica chloroplast genome as shown by isoelectric focussing of the large subunit of RUBPCase. This study demonstrated the value of N. rustica cv. chlorotica as a parental line in somatic hybridization with N. sylvestris and it might have widespread use with wild-type lines of other species.
174. E. Glotter, I. Kirson, A. Abraham, and D. Lavie, “Constituents of Withania Somnifera Dun—XIII: The Withanolides of Chemotype III,” Tetrahedron, vol. 29, no. 10, pp. 1353–1364, Jan. 1973. doi: 10.1016/S0040-4020(01)83156-2. Abstract
     Nine new steroidal lactones of the withanolide series (withanolides E-M) have been isolated from the leaves of Withania somnifera Dun (Solanaceae) growing in the southern coastal plane of Israel. This population of W. somnifera constitutes a new chemotype and is designated as chemotype III. The following structures have been assigned to seven of the above compounds: Withanolide G, 20α-hydroxy-1-oxo-20R,22R-witha-2,5,8(14),24-tetraenolide (1); withanolide H, 20α,27-dihydroxy-1-oxo-20R,22R-witha-2,5,8(14),24-tetraenolide (2); withanolide I, 20α-hydroxy-1-oxo-20R,22R-witha-3,5,8(14),24-tetraenolide (3); withanolide J, 17α,20α-dihydroxy-1-oxo-20S,22R-witha-2,5,8(14),24-tetraenolide (4); withanolide K, 17α,20α-dihydroxy-1-oxo-20S,22R-witha-3,5,8(14),24-tetraenolide (5); withanolide L, 17α,20α-dihydroxy-1-oxo-20S,22R-witha-2,5,14,24-tetraenolide (6); withanolide M, 17α,20α-dihydroxy-1-oxo-14α,15α-epoxy-20S,22R-witha-2,5,24-trienolide (7).
175. V. Goel, B. Duhan, V. K. Madan, D. E. V. Raj, and R. Prakash, “Effect of Nitrogen and FYM on Yield, Growth Parameters and Quality of Ashwagandha (Withania Somnifera),” vol. 16, pp. 101–106, Dec. 2011. Abstract
     Application of 25 mg N/kg soil and FYM 12.5 t/ha significantly increased the mean height of ashwagandha to the tune of41.6 and 8.1%, respectively, over control. Combined use of 25 mg N/kg soil and FYM @ 12.5 t/ha produced significantly taller plants (47.7 cm) in comparison to other treatments over control (31.7 cm). Application of 25 mg N/kg soil produced average number of primary branches (2.7) over control (1.5). Application of FYM @ 12.5 and 25 t/ha increased the average number of primary branches significantly from 2.0 to 2.3 and 2.6 over control. Application of 25 mg N/kg soil significantly increased the average plant spread from 134.7 to 287.6 cm2over control. The average plant spread also increased from 232.0 to 246.6 and 260.0 cm2 with the application of FYM @ 12.5 and 25 t/ha, respectively, over control. Average fresh and dry weight of ashwagandha roots increased at the tune of 17.4 and 8.9%, respectively, with the application of 37.5 mg N/kg soil. Similarly, FYM @ 25 t/ha increased the fresh and dry weight of roots to the tune-of 10.2 and 4.3%, respectively, over control. Application of 25 mg N/kg soil significantly increased mean fresh and dry weight of ashwagandha shoot to the extent of 31.6 and 31.7%, respectively, over control. Sole application of FYM @ 25 t/ha resulted in significant increase in fresh and dry weight of ashwagandha shoots to the extent of 22.2 and 20.0% over control. Application of 25 mg N/kg soil increased mean total alkaloids (%) of ashwagandha roots to the extent of 19.6% over control. Application of FYM @ 25 t/ha also increased the total alkaloid content significantly upto the extent of 14.8% over control. Similarly, application of nitrogen @ 25 mg N/kg soil increased the mean total alkaloids (%) of ashwagandha roots to the extent of 0.61% over control (0.51%) and FYM @ 25 t/ha also increased the mean alkaloids yield of ashwagandha roots from 0.54 to 0.62% over control.
176. H. Gopal, T. Natarajan, and P. Raja., “Effect of Rhizobacterial Inoculation on Rhizosphere Microflora of Ashwagandha Cv. JAWAHAR 20,” International Journal of Agricultural Sciences, vol. 2, no. 2, pp. 392–395, Jul. 2006. http://researchjournal.co.in/upload/assignments/2_392-395.pdf. Abstract
     Rhizobacteria from different medicinal plants viz., Withania somnifera, Coleus forskohlii and Vinca rosea grown in different parts of Tamil Nadu were isolated and characterized. Under pot culture conditions the rhizosphere population of Azospirillum, Azotobacter, phosphate solubilizing bacteria and Pseudomonas increased up to 120 days after inoculation and then declined. Witnessed increase in the population of respective inoculants due to biofertilizer inoculation offers more scope for plant growth promotion.
177. B. Govindaraju, R. Rao S, R. Venugopal, S. Kiran, C. Kaviraj, and S. Rao, “High Frequency Plant Regeneration in Ashwagandha [Withania Somnifera (L.) Dunal].: An Important Medicinal Plant,” Plant Cell Biotechnology & Molecular Biology, vol. 4, pp. 49–56, Mar. 2003. Abstract
     A high frequency and rapid regeneration protocol via callus and directly from various explants was developed in Withania somnifera (L.) Dunal. Callus was initiated from internodal segment, leaf, root and petiole explants on MS and B5 media supplemented with 2,4-D (0.5-3.0 mg/l) and NAA (0.5-3.0 mg/l) either alone or along with Kinetin (0.5-1.0 mg/l). Regeneration was observed from callus of all the explants except roots on MS medium fortified with BAP (0.5-2.5 mg/l) or in combination with IAA (0.5 mg/l). Direct differentiation of multiple shoots from leaf, nodal segments and shoot tips occurred within two weeks on MS medium supplemented with BAP (0.5-3.0 mg/l) in combination with IAA (0.5 mg/l). The number and height of the shoots per explant varied with different concentrations of BAP with low concentrations of IAA. Shoots that were dwarf were elongated on MS medium fortified with GA 3 (0.5 mg/l). These plantlets were then rooted successfully in half strength MS media (both liquid and solid) with IBA (0.5-1.0 mg/l) alone or along with IAA (0.5 mg/l). Plantlets were hardened for two weeks and successfully transferred to field with 80-85% survival.
178. I. Gravalos, N. Ziakas, S. Loutridis, and T. Gialamas, “A Mechatronic System for Automated Topping and Suckering of Tobacco Plants,” Computers and Electronics in Agriculture, vol. 166, p. 104986, Nov. 2019. doi: 10.1016/j.compag.2019.104986. Abstract
     In agricultural crops, such as tobacco, the budding part of the plant is removed (topping) and the growth of axillary shoots (called suckers) is controlled with chemicals in order to produce good quality heavy-bodied darker tobacco leaves. Nowadays, topping and suckering are largely performed by hand and as a result the application process is time consuming. In this study, the design and implementation of a novel mechatronic system is presented, whereby the topping and suckering treatments can be performed automatically. The mechatronic system and its auxiliary equipment were mounted on a three-wheeled mobile platform. The novel system includes a topper unit, a high-precision sprayer, and a bifurcated metal guide designed to keep the plants in an upright position. An infrared beam sensor is mounted below the circular saw blade to locate the plant stalk and trigger a pulse so that the high-precision sprayer applies the proper amount of suckercide to the tobacco stalk. The high-precision sprayer travels parallel to and over the tobacco plant rows with a speed that matches the ground speed of the wheeled mobile platform. This is achieved by the use of a built-in drive wheel encoder. The mechatronic system was tested in both laboratory and field conditions. In the tests carried out, the travel speed of the three-wheeled platform was kept between 0.1 and 0.5 m/s. The topping and suckering of five tobacco plants lasted approximately 20 s. Traditional application has a disadvantage over the mechatronic system in that each worker is independent of the others and the speed of the whole process is essentially determined by the slower worker. In future work, the mechatronic system can be integrated into a small autonomous wheeled platform, a drone, or can be mounted on the horizontal boom of a suitably modified trailer sprayer.
179. K. D. Green and N. H. Thomas, “Nicotiana Glauca: In Vitro Production of Pyridine Alkaloids and Other Secondary Metabolites,” in Medicinal and Aromatic Plants VIII, Y. P. S. Bajaj, Ed. Berlin, Heidelberg: Springer, 1995, pp. 326–343. doi: 10.1007/978-3-662-08612-4_18. Abstract
     Nicotiana glauca (family Solanaceae) is one of 21 species of Nicotiana, of which N. rustica and N. tabacum are the best known as sources of tobacco. N. glauca attains the greatest concentration and diversity in central Argentina where it is considered native. It is found less frequently in Bolivia where it also may be native. Indeed, N. glauca is represented to a greater or lesser degree in many warm temperate climates (Goodspeed 1954).
180. K. D. Green, N. H. Thomas, and J. A. Callow, “Product Enhancement and Recovery from Transformed Root Cultures of Nicotiana Glauca,” Biotechnology and Bioengineering, vol. 39, no. 2, pp. 195–202, 1992. doi: 10.1002/bit.260390211. Abstract
     Transformed roots of Nicotiana glauce synthesize the alkaloids nicotine and anabasine at levels reflecting the parent plants. Media composition, strength, and pH were evaluated with respect to biomass yield and productivity. Full-strength Gamborg’s B5 medium proved the best for biomass yield while half-strength, or low-salt, medium enhanced alkaloid accumulation. A detailed investigation of media nitrate levels demonstrated how these may be manipulated to promote growth and intracellular or extracellular alkaloid levels. High nitrate concentrations were found to significantly enhance media alkaloid levels at the end of the growth phase. Media pH is also important, although transformed roots will grow in Gamborg’s B5 medium between pH 3 and 9, root biomass is favored by an increase in medium alkalinity, while alkaloid release is encouraged by mildly acidic pH. Transformed roots release a proportion of their secondary metabolites into the growth medium. By continually removing root products, any feedback inhibition on enzymatic reactions is reduced, as are the toxic effects resulting from product accumulation. In this article we describe the use of Amberlite resins (XAD-2 and XAD-4) to enhance alkaloid levels (nicotine and anabasine) of hairy root cultures of Nicotiana glauca by a factor of 10 with no adverse effect on root growth. The performance of the Amberlite columns was subsequently investigated with respect to alkaloid adsorption and desorption, including an evaluation of the effects of pH and loading capacity. The resins also adsorb media constituents which are identified and quantified as part of this work. Resulting nutritional stresses are thought to be partly responsible for enhancing secondary metabolism at the expense of biomass yield. However, the net effects of using Amberlite resins as a means of product removal significantly increases the overall product yield and the extent to which products are released into the growth medium.
181. E. L. Griffin, G. Philips, J. Claffey, and J. Skalamera, “Nicotine Sulfate from Nicotiana Rustica,” Industrial & Engineering Chemistry, vol. 44, no. 2, pp. 274–279, Feb. 1952. doi: 10.1021/ie50506a018.
182. K. Gu et al., “The Physiological Response of Different Tobacco Varieties to Chilling Stress during the Vigorous Growing Period,” Scientific Reports, vol. 11, no. 1, p. 22136, Nov. 2021. doi: 10.1038/s41598-021-01703-7. Abstract
     Tobacco is be sensitively affected by chilling injury in the vigorous growth period, which can easily lead to tobacco leaf browning during flue-curing and quality loss, however, the physiological response of tobacco in the prosperous period under low temperature stress is unclear. The physiological response parameters of two tobacco varieties to low temperature stress were determined. The main results were as follows: ① For tobacco in the vigorous growing period subjected to low-temperature stress at 4–16 °C, the tissue structure of chloroplast changed and photosynthetic pigments significantly decreased compared with each control with the increase of intensity of low-temperature stress. ② For tobacco in the vigorous growing period at 10–16 °C, antioxidant capacity of the protective enzyme system, osmotic adjustment capacity of the osmotic adjusting system and polyphenol metabolism in plants gradually increased due to induction of low temperature with the increase of intensity of low-temperature stress. ③ Under low-temperature stress at 4 °C, the protective enzyme system, osmotic adjusting system and polyphenol metabolism of the plants played an insignificant role in stress tolerance, which cannot be constantly enhanced based on low-temperature resistance at 10 °C. This study confirmed that under the temperature stress of 10–16 °C, the self-regulation ability of tobacco will be enhanced with the deepening of low temperature stress, but there is a critical temperature between 4 and 10 °C. The self-regulation ability of plants under low temperature stress will be inhibited.
183. GuidanceGD, “Ashwagandha Harvest and Processing. Indian Ginseng Root and Seed Collection.” Apr-2021. https://www.youtube.com/watch?v=F4fkf0QkhhM. Abstract
     #Ashwagandha #ginsengRoot Ashwagandha harvest and processing. Root and Seed collection. ginseng root. Indian ginseng root. Ginseng root cultivation. Ginseng refers to eleven different varieties of a short, slow-growing plant with fleshy roots. Ginseng is believed to restore and enhance wellbeing. It is one of the most popular herbal remedies. Ginseng has traditionally been taken to aid a range of medical conditions. #Ashwagandha #Medicinal_herb In rural areas of India , farmer are cultivating medicinal herbs and getting high profit than other crops. Here in this video you will see how the farmers of West Bengal are cultivating indian ginseng i.e ashwagandha and becoming economically stable. Ashwagandha or indian ginseng ( Withaniasomnifera) is a highly effective ayurvedic medicine. There are many ayurvedic medicines which are prepared by Ashwagandha. The demand and supply of ashwagandha is not equal in the market that’s why the price is always high. The avarage price of dried ashwagandha root in the market is Rs. 25000 /- to Rs. 40000/-. So those farmer who want to earn high profit from cultivation may take a chance in ashwagandha. More research is needed to confirm its benefit as a supplement. However, it is claimed that ginsenosides, chemical components found in ginseng, are responsible for the clinical effect of the herb.
184. X. Guo, J. Niu, and X. Cao, “Heterologous Expression of Salvia Miltiorrhiza MicroRNA408 Enhances Tolerance to Salt Stress in Nicotiana Benthamiana,” International Journal of Molecular Sciences, vol. 19, no. 12, p. 3985, Dec. 2018. doi: 10.3390/ijms19123985. Abstract
     MicroRNAs (miRNAs) are a class of endogenous small RNAs that regulate the expression of target genes post-transcriptionally; they are known to play major roles in development and responses to abiotic stress. MicroRNA408 (miR408) is a conserved small RNA in plants; it was reported that miR408 genes were involved in abiotic stress in Arabidopsis. However, miR408 in Salvia miltiorrhiza has been rarely investigated. In this study, we cloned Sm-MIR408, the miR408 precursor sequence, and its promoter sequence from S. miltiorrhiza and the role in tolerance to salt stress is described. The effects of salt stress on miR408 expression were studied by using β-glucuronidase (GUS) staining. Our data indicated that transgenic tobacco overexpressing Sm-MIR408 promoted seed germination and reduced the accumulation of reactive oxygen species under salt stress. Transcript levels of antioxidative genes, i.e., NbSOD, NbPOD, and NbCAT, and their enzyme activities increased in salinity-stressed transgenic tobacco plants, suggesting a better antioxidant system to cope the oxidative damage caused by salinity stress. Taken together, these findings indicated that miR408 functions in positive responses to salt tolerance in tobacco.
185. R. Gupta, A. Singh, M. Srivastava, M. M. Gupta, and R. Pandey, “Augmentation of Systemic Resistance and Secondary Metabolites by Chitinolytic Microbes in Withania Somnifera against Meloidogyne Incognita,” Biocontrol Science and Technology, vol. 26, no. 12, pp. 1626–1642, Dec. 2016. doi: 10.1080/09583157.2016.1230729. Abstract
     Ashwagandha (Withania somnifera L. Dunal), a therapeutically imperative herb is known for its useful steroidal lactones (withanolide and withaferin) and acyl steryl glycosides. The plant is a highly susceptible host for the plant parasitic nematode, Meloidogyne incognita. Selected chitinolytic microbes, namely Cellulosimicrobium cellulans MTN13, Flavobacterium johnsoniae MTN 20, Chitiniphilus sp. MTN22 and Streptomyces sp. MTN14, alone and in combination for M. incognita management and enhancement of secondary metabolites in W. somnifera cv. Poshita were evaluated. A significant enhancement in biomass yield (1.9-fold) and disease diminution (2.7-fold) was found in the dual microbial treatment Streptomyces sp. and Chitiniphilus sp. with respect to the untreated inoculated control plants. A significant augmentation (2.7- and 2.0-fold, respectively) in withanolide A and withaferin A was also found in the same treatment. The studies revealed increment of defence variables 1.1- to 1.3-fold and 1.6- to 2.1-fold in single- and dual-microbe treatments, respectively, than the untreated inoculated plants. The stimulation of the phenylpropanoid pathway and phenolics accumulation was the maximum at 5 days post inoculation (dpi), whereas antioxidant enzymes activities were the highest at 7 dpi. The results thus highlight a possible new function of chitinolytic microbes alone and in combinations that can effectively manage M. incognita-induced stress along with enhanced active molecules of W. somnifera.
186. P. Gupta, A. V. Agarwal, N. Akhtar, R. S. Sangwan, S. P. Singh, and P. K. Trivedi, “Cloning and Characterization of 2-C-Methyl-d-Erythritol-4-Phosphate Pathway Genes for Isoprenoid Biosynthesis from Indian Ginseng, Withania Somnifera,” Protoplasma, vol. 250, no. 1, pp. 285–295, Feb. 2013. doi: 10.1007/s00709-012-0410-x. Abstract
     Withania somnifera (L.) is one of the most valuable medicinal plants used in Ayurvedic and other indigenous medicines. Pharmaceutical activities of this herb are associated with presence of secondary metabolites known as withanolides, a class of phytosteroids synthesized via mevalonate (MVA) and 2-C-methyl-d-erythritol-4-phosphate pathways. Though the plant has been well characterized in terms of phytochemical profiles as well as pharmaceutical activities, not much is known about the genes responsible for biosynthesis of these compounds. In this study, we have characterized two genes encoding 1-deoxy-d-xylulose-5-phosphate synthase (DXS; EC 2.2.1.7) and 1-deoxy-d-xylulose-5-phosphate reductase (DXR; EC 1.1.1.267) enzymes involved in the biosynthesis of isoprenoids. The full-length cDNAs of W. somnifera DXS (WsDXS) and DXR (WsDXR) of 2,154 and 1,428 bps encode polypeptides of 717 and 475 amino acids residues, respectively. The expression analysis suggests that WsDXS and WsDXR are differentially expressed in different tissues (with maximal expression in flower and young leaf), chemotypes of Withania, and in response to salicylic acid, methyl jasmonate, as well as in mechanical injury. Analysis of genomic organization of WsDXS shows close similarity with tomato DXS in terms of exon–intron arrangements. This is the first report on characterization of isoprenoid biosynthesis pathway genes from Withania.
187. P. Gupta et al., “Comparative Transcriptome Analysis of Different Chemotypes Elucidates Withanolide Biosynthesis Pathway from Medicinal Plant Withania Somnifera,” Scientific Reports, vol. 5, no. 1, p. 18611, Dec. 2015. doi: 10.1038/srep18611. Abstract
     Withania somnifera is one of the most valuable medicinal plants synthesizing secondary metabolites known as withanolides. Despite pharmaceutical importance, limited information is available about the biosynthesis of withanolides. Chemo-profiling of leaf and root tissues of Withania suggest differences in the content and/or nature of withanolides in different chemotypes. To identify genes involved in chemotype and/or tissue-specific withanolide biosynthesis, we established transcriptomes of leaf and root tissues of distinct chemotypes. Genes encoding enzymes for intermediate steps of terpenoid backbone biosynthesis with their alternatively spliced forms and paralogous have been identified. Analysis suggests differential expression of large number genes among leaf and root tissues of different chemotypes. Study also identified differentially expressing transcripts encoding cytochrome P450s, glycosyltransferases, methyltransferases and transcription factors which might be involved in chemodiversity in Withania. Virus induced gene silencing of the sterol ∆7-reductase (WsDWF5) involved in the synthesis of 24-methylene cholesterol, withanolide backbone, suggests role of this enzyme in biosynthesis of withanolides. Information generated, in this study, provides a rich resource for functional analysis of withanolide-specific genes to elucidate chemotype- as well as tissue-specific withanolide biosynthesis. This genomic resource will also help in development of new tools for functional genomics and breeding in Withania.
188. P. Gupta, N. Akhtar, S. K. Tewari, R. S. Sangwan, and P. K. Trivedi, “Differential Expression of Farnesyl Diphosphate Synthase Gene from Withania Somnifera in Different Chemotypes and in Response to Elicitors,” Plant Growth Regulation, vol. 65, no. 1, pp. 93–100, Sep. 2011. doi: 10.1007/s10725-011-9578-x. Abstract
     Withania somnifera (L.) Dunal (Family, Solanaceae), commonly known as Ashwagandha is one of the most valuable medicinal plants synthesizing large number of pharmacologically active secondary metabolites known as withanolides. Though the plant has been well characterized in terms of phytochemical profiles as well as pharmaceutical activities, not much is known about the genes responsible for biosynthesis of these compounds. In this study, we have characterized a gene encoding farnesyl diphosphate synthase (FPPS; EC 2.5.1.10), a key enzyme in the pathway of biosynthesis of isoprenoids, from W. somnifera. The full-length cDNA of Withania somnifera FPPS (WsFPPS) of 1,253 bps encodes a polypeptide of 343 amino acids. The amino acid sequence homology and phylogenetic analysis suggest that WsFPPS has close similarity to its counterparts from tomato (SlFPPS) and capsicum (CaaFPPS). Using semi quantitative RT–PCR, the expression pattern of the WsFPPS gene was analyzed in different tissues of Withania chemotypes (NMITLI-101, NMITLI-108, NMITLI-118 and NMITLI-135) as well as in response to elicitors (salicylic acid and methyl jasmonate) and mechanical wounding. The expression analysis suggests that WsFPPS expression varies in different tissues (with maximal expression in flower and young leaf) and chemotypes (with highest level in NMITLI-101) and was significantly elevated in response to salicylic acid, methyl jasmonate and mechanical injury. This is the first report on characterization of an isoprenoid pathway gene involved in withanolide biosynthesis.
189. B. M. Gupta and K. K. M. Ahmed, “Research on Withania Somnifera (Ashwaganda): A Scientometric Assessment of Global Publications Output during 1995-2018,” International Journal of Pharmaceutical Investigation, vol. 9, no. 2, pp. 59–66, Sep. 2019. doi: 10.5530/ijpi.2019.2.12. Abstract
     The present study examined 2008 global publications on Withania somnifera, as indexed and covered in international Scopus database during 1995-18, with a view to understand their publications growth rate, global publication share, citation impact, share of their international collaborative publications, distribution of publications by broad subjects, productivity and citation profile of top organizations and authors, preferred media of communication and bibliographic characteristics of high cited publications. The global publications registered 18.31% annual average growth rate and its citation impact averaged to 18.55 citations per publication. The global share of top 10 countries ranged from 1.34% to 70.42%, with the largest share (70.42%) from India, followed by USA (11.35%) and other 8 countries from 1.34% to 4.03%. More than 100% of the cumulative global publication and citation share comes from top 10 countries during 1995-18. Only four countries among top 10 registered relative citation index above the world average of 1.0: U.K. (2.48), USA (1.80), Germany (1.69) and South Korea (1.08) during 1995-18. Pharmacology, toxicology and pharmaceutics contributed the largest global publications share of 42.33%, followed biochemistry, genetics and molecular biology (32.42%), medicine (30.73%), agricultural & biological sciences (25.45%), etc. during 1995-18. 377 global organizations and 422 global authors participated in global Withania somnifera research, of which the 15 most productive global organizations and authors together contributed 25.65% and 14.59% global publication share and 32.31% and 23.82% global citation share respectively during 1995-18. Amongst 1964 journal publications (in 313 journals) in global Withania somnifera research, the top 15 most productive journals together contributed 21.49% global share of total journal publication output during 1995-18. Sixty Eight (68) publications were found to be high cited, as they registered citations from 100 to 601 during 1995-18 and they together received 11582 citations, averaging to 170.32 citations per publication.
190. M. L. Gupta, H. O. Misra, A. Kalra, and S. P. S. Khanuja, “Root-Rot and Wilt: A New Disease of Ashwagandha (Withania Somnifera) Caused by Fusarium Solani.,” Journal of Medicinal and Aromatic Plant Sciences, vol. 26, no. 2, pp. 285–287, 2004. https://www.cabdirect.org/cabdirect/abstract/20053051122. Abstract
     Symptoms of root rot and wilt were observed in the fields of ashwagandha (W. somnifera) in Lucknow (Uttar Pradesh, India) conditions. Initial symptoms were withering and drooping of the plants while at later stages, plants showed severe wilting leading to death and decay of underground parts. The root of infected plant showed pulpiness with brownish colour. White cottony growth of the fungus was...
191. M. Gupta, P. K. Srivastava, Shikha, A. Niranjan, and S. K. Tewari, “Use of a Bioaugmented Organic Soil Amendment in Combination with Gypsum for Withania Somnifera Growth on Sodic Soil,” Pedosphere, vol. 26, no. 3, pp. 299–309, Jun. 2016. doi: 10.1016/S1002-0160(15)60044-3. Abstract
     Limited availability of organic matter is a problem to sustain crop growth on sodic soil. Organic soil amendments are a cost-effective source of nutrients to enhance crop growth. A field study was conducted to evaluate the effect of an organic soil amendment bioaugmented with plant growth-promoting fungi (SFOA) in combination with gypsum on soil properties and growth and yield attributes of Withania somnifera, one of the most valuable crops of the traditional medicinal system in the world, on a sodic soil at the Aurawan Research Farm of CSIR-National Botanical Research Institute, Lucknow, India. The SFOA used was prepared by pre-enriching farm waste vermicompost with plant growth-promoting fungi before mixing with pressmud and Azadirachta indica seed cake. The application of SFOA at 10 Mg ha−1 after gypsum (25.0 Mg ha−1) treatment significantly (P < 0.05) increased root length (by 96%) and biomass (by 125%) of Withania plants compared to the control without SFOA and gypsum. Similarly, the highest withanolide contents were observed in leaves and roots of Withania plants under 10 Mg ha−1 SFOA and gypsum. Combined application of SFOA and gypsum also improved physical, chemical and enzymatic properties of the soil, with the soil bulk density decreasing by 25%, water-holding capacity increasing by 121%, total organic C increasing by 90%, pH decreasing by 17% and alkaline phosphatase, β-glucosidase, dehydrogenase and cellulase activities increasing by 54%, 128%, 81% and 96%, respectively, compared to those of the control. These showed that application of the SFOA tested in this study might reclaim sodic soil and further support Withania cultivation and results were better when the SFOA was applied after gypsum treatment.
192. G. Gupta, A. C. Rana, A. C. Rana, and A. C. Rana, “Withania Somnifera (Ashwagandha): A Review,” Pharmacognosy Reviews, vol. 1, no. 1, pp. 129–136, 2007.
193. J. W. Ha et al., “Structural Characterization of Withanolide Glycosides from the Roots of Withania Somnifera and Their Potential Biological Activities,” Plants, vol. 11, no. 6, p. 767, Jan. 2022. doi: 10.3390/plants11060767. Abstract
     Withania somnifera (Solanaceae), commonly known as “ashwagandha”, is an ayurvedic medicinal plant that has been used for promoting good health and longevity. As part of our ongoing natural product research for the discovery of bioactive phytochemicals with novel structures, we conducted a phytochemical analysis of W. somnifera root, commonly used as an herbal medicine part. The phytochemical investigation aided by liquid chromatography-mass spectrometry (LC/MS)-based analysis led to the isolation of four withanolide glycosides (1–4), including one new compound, withanoside XII (1), from the methanol (MeOH) extract of W. somnifera root. The structure of the new compound was determined by nuclear magnetic resonance (NMR) spectroscopic data, high-resolution (HR) electrospray ionization (ESI) mass spectroscopy (MS), and electronic circular dichroism (ECD) data as well as enzymatic hydrolysis followed by LC/MS analysis. In addition, enzymatic hydrolysis of 1 afforded an aglycone (1a) of 1, which was identified as a new compound, withanoside XIIa (1a), by the interpretation of NMR spectroscopic data, HR-ESIMS, and ECD data. To the best of our knowledge, the structure of compound 2 (withagenin A diglucoside) was previously proposed by HRMS and MS/MS spectral data, without NMR experiment, and the physical and spectroscopic data of withagenin A diglucoside (2) are reported in this study for the first time. All the isolated compounds were evaluated for their anti-Helicobacter pylori, anti-oxidant, and anti-inflammatory activities. In the anti-Helicobacter pylori activity assay, compound 2 showed weak anti-H. pylori activity with 7.8% inhibition. All the isolated compounds showed significant ABTS radical scavenging activity. However, all isolates failed to show inhibitory activity against nitric oxide (NO) production in lipopolysaccharide-stimulated RAW 264.7 macrophage cells. This study demonstrated the experimental support that the W. somnifera root is rich in withanolides, and it can be a valuable natural resource for bioactive withanolides.
194. R. Hajiboland and N. Ebrahimi, “Effect of Exposure to Uv Radiation on Growth, Photosynthesis and Antioxidant Defense System in Tobacco (Nicotiana Rustica L. Cv. Basmas) Plants Treated with Exogenous Polyamines.,” Genetics and Plant Physiology, vol. 1, no. 1/2, pp. 76–90, 2011. https://www.cabdirect.org/cabdirect/abstract/20133089643. Abstract
     Tobacco (Nicotiana rustica L. cv. Basmas) plants were exposed to a daily dose of 10 kJ m-1d-1 of UV radiation and treated with 0.5 mM exogenous polyamines (PAs) applied to the nutrient solution for two weeks. Dry weight of leaves increased under UVAB treatment in the absence of PAs and presence of spermidine but not putrescine. In the presence of putrescine, UVA treatment caused a significant...
195. R. Hajiboland, L. Cheraghvareh, and C. Poschenrieder, “Improvement of Drought Tolerance in Tobacco (Nicotiana Rustica L.) Plants by Silicon,” Journal of Plant Nutrition, vol. 40, no. 12, pp. 1661–1676, Jul. 2017. doi: 10.1080/01904167.2017.1310887. Abstract
     Ameliorative effect of silicon (Si) (2 mM as sodium silicate (Na2SiO3)) was studied in tobacco (Nicotiana rustica L.) plants grown under control at 100% field capacity (FC), mild drought (60% FC), and severe drought (30% FC) conditions. Si-treated plants had higher biomass of particularly above-ground parts both under drought and control conditions. Plants with Si supply had significantly higher net assimilation rates but lower transpiration rates. Silicon supply enhanced osmotic potentials only in the leaves, but not in the roots. A considerable rise in the concentrations of soluble sugars was observed particularly in the leaves under both drought and Si treatments. Soluble proteins, free α-amino acids, and proline concentrations increased in Si-treated plants under all watering treatments. Si enhanced the activity of antioxidative enzymes and decreased hydrogen peroxide (H2O2) concentrations. Results indicate that Si supplementation alleviates drought stress via improvement of water relation parameters, enhancement of photosynthesis, and elevation of antioxidant defenses.
196. R. Hajiboland and L. Cheraghvareh, “Influence of Si Supplementation on Growth and Some Physiological and Biochemical Parameters in Salt-Stressed Tobacco (Nicotiana Rustica L.) Plants,” Journal of Sciences, Islamic Republic of Iran, vol. 25, no. 3, pp. 205–217, Sep. 2014. https://jsciences.ut.ac.ir/article_51849.html. Abstract
     Tobacco is a salt-sensitive glycophyte crop species. In this work effect of silicone (Si) supplementation (1 mM as Na2SiO3) was studied in Nicotiana rustica L. cv. Basmas grown hydroponically in growth chamber under control, low (25 mM) and high (75 mM) NaCl concentration for two weeks. Dry matter production of leaves was depressed by salinity level as low as 25 mM and higher salt concentration decreased plants dry weight by 52-82%. Si supplementation alleviated salt stress effect as could be judged by higher dry weight of shoot and roots in +Si plants compared with –Si counterparts. Leaf chlorophyll a and carotenoids concentrations and net assimilation rate were higher in Si-treated plants not only in salt-affected but also in control plants. Si treatment resulted in higher concentration of soluble carbohydrates but not proline. Leaf transpiration rate, unexpectedly, was not diminished by Si and water use efficiency was rather lowered by Si in salt-treated plants. Si application caused a slight reduction of Na concentration while increased that of K and Ca significantly and resulted in higher K:Na ratio in the leaves, stem and roots. Our results suggested that Si application improved tolerance to salt stress in tobacco due to an enhancement of photosynthesis, accumulation of organic osmolytes as well as improvement of K:Na selectivity but not limiting water loss. In addition, greater dry matter production of Si-supplemented plants in the absence of salt was associated with elevated photosynthesis rate, higher K and Ca uptake and proline content.
197. D. E. Haley, F. D. Gardner, and R. T. Whitney, “Nicotiana Rustica as a Source of Nicotine for Insect Control,” Science, Oct. 1924. https://www.science.org/doi/abs/10.1126/science.60.1555.365.
198. D. E. Haley, O. Olson, and F. L. Follweiler, “Studies on Nicotiana Rustica as a Source of a Nicotine for Insect Control,” Journal of Economic Entomology, vol. 18, no. 6, pp. 807–817, Dec. 1925. doi: 10.1093/jee/18.6.807. Abstract
     This investigation had for its purpose the testing of the rate of volatilization of nicotine from ground Nicotiana Rustica (a high nicotine tobacco) in order to ascertain whether or not this type of tobacco could be used as an insecticidal dust.The ground tobacco was used alone, or with other materials.The rate of nicotine evolution was measured by the death rate or suspended animation of cabbage aphids (Aphis brassicae Linnaeus) and other soft-bodied insects.As a result of these studies the following conclusions seem justified:(1) The ground tobacco, when used alone, with water or with basic lime compounds was not efficient as an insecticide in the control of aphids.(2) If sufficient quantities of basic materials were mixed with the ground tobacco, it was found that the addition of water resulted in the evolution of nicotine, the rate of evolution depending in a large measure on the amount of water added.(3) A mixture containing 5 parts of tobacco, 5 parts of hydrated lime and 3 parts of water proved to be very efficient as an insecticide under laboratory and field conditions. Notwithstanding the relatively large amount of water present, it was found that such a mixture could be applied to vegetation by the use of power blowers and with no especial difficulty.(4) Mixtures of the ground tobacco, water and basic lime compounds could be kept in sealed containers for a relatively long period of time with no apparent loss of toxicity.(5) These experiments warrant that some consideration be given to the possibility of cultivating Nicotiana Rustica for its nicotine content alone and to utilize it in the form of an insecticidal dust.
199. J. D. Hamill, D. Pental, and E. C. Cocking, “Analysis of Fertility in Somatic Hybrids of Nicotiana Rustica and N. Tabacum and Progeny over Two Sexual Generations,” Theoretical and Applied Genetics, vol. 71, no. 3, pp. 486–490, Dec. 1985. doi: 10.1007/BF00251193. Abstract
     Somatic hybrid plants, produced between Nicotiana rustica and N. tabacum by heterokaryon isolation and culture and also by mutant complementation, were examined regarding their ability to set seed. From a total of seventeen independent somatic hybrids, three were found to be partially self-fertile while the others did not set seed. Differences regarding the methods of hybrid selection, parental varieties and chloroplast composition of hybrids did not appear to be significant regarding the ability of plants to set seed. Much variation in fertility was observed in subsequent generations and by recurrent selection of the most fertile, over two generations, it was possible to increase the level of self-fertility in some of the progeny. One R2 derivative possessed approximately a tenfold higher level of self-fertility than it’s somatic hybrid parent. The presence of genetic markers from both parents were observed in all progeny indicating their hybrid nature.
200. J. D. Hamill, A. J. Parr, R. J. Robins, and M. J. C. Rhodes, “Secondary Product Formation by Cultures of Beta Vulgaris and Nicotiana Rustica Transformed with Agrobacterium Rhizogenes,” Plant Cell Reports, vol. 5, no. 2, pp. 111–114, Apr. 1986. doi: 10.1007/BF00269247. Abstract
     ‘Hairy root’ cultures of Beta vulgaris and Nicotiana rustica were established after roots were induced on plants following infection with Agrobacterium rhizogenes. The transformed cultures of B. vulgaris and N. rustica synthesised their characteristic secondary products, the betalain pigments and nicotine alkaloids respectively, at levels comparable with those of in vivo roots from the same variety. Betalains were entirely retained inside the root tissue. In contrast, a proportion of the nicotine alkaloids was secreted into the medium. The potential of this type of ‘in vitro’ plant tissue culture for the production of valuable plant secondary products is identified and confirmed.
201. A. K. Handique and C. M. Sharma, “Colchicine Induced Hypertrophy in Catharanthus Roseus (L.) G. Don and Nicotiana Rustica L.,” Crop Research (Hisar), vol. 3, no. 2, pp. 287–290, 1990. https://www.cabdirect.org/cabdirect/abstract/19920315522. Abstract
     Dry seeds of Catharanthus roseus and Nicotiana rustica were surface sterilized with 0.01% HgCl2. The seeds were then treated with colchicine (0.1, 0.2, 0.3 or 0.4%) for 8, 16 or 24 h. Hypertrophy was observed in the epicotyl, and the diameter of the middle of the epicotyl was used to determine the degree of hypertrophy. Epicotyl diameter increased with increasing colchicine concentration. In C....
202. T. Hartmann, L. Witte, F. Oprach, and G. Toppel, “Reinvestigation of the Alkaloid Composition of Atropa Belladonna Plants, Root Cultures, and Cell Suspension Cultures,” Planta Medica, vol. 52, no. 5, pp. 390–395, Oct. 1986. doi: 10.1055/s-2007-969194. Abstract
     Thieme E-Books & E-Journals
203. E. Heberle-Bors, “Isolated Pollen Culture in Tobacco: Plant Reproductive Development in a Nutshell,” Sexual Plant Reproduction, vol. 2, no. 1, pp. 1–10, Mar. 1989. doi: 10.1007/BF00190112. Abstract
     In the past, in vitro cultures of excised anthers and isolated pollen have been used to study normal male sexual development (gametophytic development) and, conversely, to produce and study haploid plant formation (sporophytic development). For years both branches have existed side by side, without much interaction. Today, a synthesis of the two branches is possible as well as necessary. Recent advances in the technique of isolated pollen culture in the tobacco plant model (Nicotiana tabacum L.) enable the researcher to strictly control pollen development in both the gametophytic and sporophytic direction. The nutritional status of the immature pollen grain at a particular stage of development provides the trigger for its development into one of the two phases found in the alternation of generations undergone by higher plants. In particular, a hunger signal is responsible for the derepression of cell division activity and the start of embryogenesis. Pollen starvation can occur in isolated pollen cultures in sucrose-free media, in excised anthers and flowers, and, under specific growth conditions, during pollen development in vivo.
204. HerbalGram - American Botanical Council, “Medicine Hunter Presents: Ashwagandha Harvest.” Sep-2017. https://www.youtube.com/watch?v=SnSWLvddqxM. Abstract
     Our mission is to provide education using science-based and traditional information to promote responsible use of herbal medicine — serving the public, researchers, educators, healthcare professionals, industry and media. Disclaimer: This video was produced independently by Chris Kilham. Information provided is for educational purposes. The views expressed by all contributors belong to them and do not necessarily reflect the views of the American Botanical Council.
205. T. Hidayat, D. Priyandoko, P. Y. Wardiny, and D. K. Islami, “Molecular Phylogenetic Screening of Withania Somnifera Relative From Indonesia Based on Internal Transcribed Spacer Region,” HAYATI Journal of Biosciences, vol. 23, no. 2, pp. 92–95, Apr. 2016. doi: 10.1016/j.hjb.2016.02.002. Abstract
     Withania somnifera (family Solanaceae), known commonly as Ashwaganda, is one of the important medicinal plants, and recent studies reported that Withanone, one of the chemical components in this plant, has ability to kill cancer cell. Because of endemic state of this plant to South Asia, exploring plant species under the same family which grow well in Indonesia has been of interest. The purpose of this study was to screen the Indonesian plant which has strong phylogenetic relationship with Ashwaganda. Thus, phylogenetic analysis using DNA sequences of internal transcribed spacer (ITS) region was conducted. Thus, 19 species of Solanaceae and two species of Convolvulaceae as outgroup were examined. Five ITS regions of Ashwaganda retrieved from GenBank were included in the phylogenetic analysis. Parsimony analysis showed that Indonesia Solanaceae comprises seven groups which is consistent with the global Solanaceae relationship as previously reported. Furthermore, our study revealed that two species, Physalis angulata and Physalis peruviana, are relative to W. somnifera. Morphologically, they share characters of flower and fruit. This result indicated that these two species are potential to have similar chemical properties as Ashwaganda, thus we can have new variants of Withanone originated from Indonesia with similar effect.
206. J. Hill, “The Environmental Induction of Heritable Changes in NICOTIANA RUSTICA Parental and Selection Lines,” Genetics, vol. 55, no. 4, pp. 735–754, Apr. 1967. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1211475/.
207. J. Hill and J. M. Perkins, “The Environmental Induction of Heritable Changes in NICOTIANA RUSTICA. Effects of Genotype-Environment Interactions,” Genetics, vol. 61, no. 3, pp. 661–675, Mar. 1969. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1212232/.
208. A. Hlasnikova and J. Dubovsky, “Androgenesis in Vitro and Associated Problems. V. Comparison of the Effect of Saccharose and Glucose on the Pollen Germination in Some Sorts and Varieties of Nicotiana.,” Acta Facultatis Rerum Naturalium Universitatis Comenianae, Genetica, pp. 11–20, 1980. https://www.cabdirect.org/cabdirect/abstract/19810725339. Abstract
     Pollen of (a) 3 tobacco cv., (b) Nicotiana rustica and (c) N. alata was germinated in a liquid medium containing 0, 1, 3, 9, 12, 15, 18, 21 or 24% of glucose or sucrose. Pollen germination and pollen tube length were higher with sucrose than with glucose in (a), but in (b) and (c) the differences were non-significant and decreased with increasing conc. of both sugars.
209. M. I. C. H. A. E. L. C. HOBBS and M. I. C. H. A. E. L. M. YEOMAN, “Effect of Light on Alkaloid Accumulation in Cell Cultures of Nicotiana Species,” Journal of Experimental Botany, vol. 42, no. 11, pp. 1371–1378, Nov. 1991. doi: 10.1093/jxb/42.11.1371. Abstract
     The effect of light on alkaloid accumulation in a range of cell cultures of tobacco was determined. Cell suspension cultures of Nicoriana rabacwn L. cv. Wisconsin-38 with differing degrees of photosynthetic activity, callus cultures of N. glauca Graham, root cultures of N. rustica L. and shoot cultures of N. tabacum were used. The alkaloid content of green illuminated cultures was greatly reduced compared with non-green cultures grown in the dark, but decreased accumulation did not correlate with increasing photosynthetic activity. The accumulation of all of the major alkaloids was affected, regardless of the species of tobacco used. Transfer of N. glauca callus from the dark into the light caused a decrease in alkaloid accumulation, while moving cultures from the light into the dark resulted in an increase in alkaloid content. In root cultures light caused a reduction in growth, which affected alkaloid synthesis. In shoot cultures there were only traces of alkaloid detectable, regardless of whether or not cultures were illuminated. Light appeared to cause a non-photosynthetic suppression of alkaloid accumulation in visibly undifferentiated cultures, and this effect was modified in visibly differentiated cultures.
210. P. A. Hosamani, H. C. Lakshman, K. Sandeepkumar, M. A. Kadam, and A. S. Kerur, “Role of Arbuscular Mycorrhizae in Conservation of Withania Somnifera.,” Bioscience Discovery Journal, vol. 2, no. 2, pp. 201–206, 2011. https://www.cabdirect.org/cabdirect/abstract/20113222949. Abstract
     An experiment was undertaken to study the selected medicinal plant and their therapeutic value with the association of Arbuscular mycorrhizal fungi. Rhizospheric soil samples were collected from the local places of Dharwad, which is geographically located between 14°15′ and 15°50′ North longitudes and 74°48′ and 76°20′ east latitude of northwestern part of Karnataka state, India. The present...
211. J.-P. Huang, Y.-J. Wang, T. Tian, L. Wang, Y. Yan, and S.-X. Huang, “Tropane Alkaloid Biosynthesis: A Centennial Review,” Natural Product Reports, vol. 38, no. 9, pp. 1634–1658, Sep. 2021. doi: 10.1039/D0NP00076K. Abstract
     Covering: 1917 to 2020 Tropane alkaloids (TAs) are a remarkable class of plant secondary metabolites, which are characterized by an 8-azabicyclo[3.2.1]octane (nortropane) ring. Members of this class, such as hyoscyamine, scopolamine, and cocaine, are well known for their long history as poisons, hallucinogens, and anaesthetic agents. Since the structure of the tropane ring system was first elucidated in 1901, organic chemists and biochemists have been interested in how these mysterious tropane alkaloids are assembled in vitro and in vivo. However, it was only in 2020 that the complete biosynthetic route of hyoscyamine and scopolamine was clarified, and their de novo production in yeast was also achieved. The aim of this review is to present the innovative ideas and results in exploring the story of tropane alkaloid biosynthesis in plants from 1917 to 2020. This review also highlights that Robinson’s classic synthesis of tropinone, which is one hundred years old, is biomimetic, and underscores the importance of total synthesis in the study of natural product biosynthesis.
212. W. F. Hunt and R. S. Loomis, “Carbohydrate-Limited Growth Kinetics of Tobacco (Nicotiana Rustica L.) Callus 1,” Plant Physiology, vol. 57, no. 5, pp. 802–805, May 1976. doi: 10.1104/pp.57.5.802. Abstract
     Logistic curves were fitted to sigmoidal growth data obtained from tobacco (Nicotiana rustica L.) callus grown on media prepared with 0.1, 0.03, 0.01, and 0.003 m sucrose. Analysis of the growth curves indicated that final yields and specific growth rates were influenced by the initial sucrose concentration. Growth yields from the four treatments were similar (0.61 ± 0.04 gram dry tissue per gram sucrose supplied). Initial specific growth rates exhibited a Michaelis-Menten dependency on initial sucrose concentration such that the Vmax = 0.18 g g−1 day−1 and Km = 0.0037 m sucrose.
213. M. Iqbal and A. K. Datta, “Ashwagandha (Withania Somnifera) - the Elixir of Life.,” Plant Archives, vol. 7, no. 2, pp. 449–455, 2007. https://www.cabdirect.org/cabdirect/abstract/20083026924. Abstract
     Withania somnifera (Solanaceae) is commonly known as Ashwagandha or Asgandh. It is a shrubby bush that grows in dry and arid soils of subtropical regions. Ashwagandh is mainly cultivated for its medicinal roots. Many biochemically heterogeneous alkaloids have been reported in the roots and the principle among them are cuscohyrine, anahygrine, tropine, pseudotropine, anaferine, di-isopelleteirine,...
214. M. Iqbal and A. K. Datta, “Induced Mutagenesis in Withania Somnifera.,” Journal of Tropical Medicinal Plants, vol. 8, no. 1, pp. 47–53, 2007. https://www.cabdirect.org/cabdirect/abstract/20073177454. Abstract
     Ten morphological mutants of Withania somnifera were isolated at M2 following gamma irradiation and hydroxylamine and EMS treatments to dry seeds (Ashwagandha; family: Solanaceae). Viable mutation frequency was highest for hydroxylamine (21.18%) followed by gamma irradiation (18.86%) and EMS (17.65%). Thick stem I and II, bushy, and ovate leaf mutants were promising for few phenotypic traits. The...
215. M. Iqbal and A. K. Datta, “A Report on a Paracentric Inversion in Withania Somnifera (L.) Dun.,” INDIAN JOURNAL OF GENETICS AND PLANT BREEDING, vol. 66, no. 01, pp. 73–74, Feb. 2006. https://www.isgpb.org/journal/index.php/IJGPB/article/view/1464. Abstract
     Cytogenetical studies in Withania somnifera (L.) Dun., an important evergreen tomentose shrub belonging to the family Solanaceae with immense therapeutic uses including anticancerous [1] and antioxidant [2] properties, is restricted to chromosome counts only and has been reported to be variable as 2n = 24 [3], 48 [4] and 72 [5]. From the seed stock of Withania somnifera (L.) Dun. (Neemuch cultivar : MPST No. NH-II/ XVI 11/946, obtained from Agri-Horticultural Society of India) plants were raised in the experimental field of Kalyani University and a plant possessing characteristic configurations of paracentric inversion [6] at anaphase I and II was identified (1 out of 26 plants scored cytologically) and has been described in the text.
216. Ö. D. A. R. C. A. N. S. O. Y. İşeri, D. A. K. S. O. Y. Körpe, and F. I. Sahin, “High Salt Induced Oxidative Damage and Antioxidant Response Differs in Nicotiana Tabacum L. and Nicotiana Rustica L. Cultivars,” Journal of Applied Biological Sciences, vol. 7, no. 1, pp. 15–22, 2013. http://jabsonline.org/index.php/jabs/article/view/318. Abstract
     We investigated effects of 10-day irrigation with 0.2M, 0.4M, and 0.6M NaCl on two tobacco variants cultivated in Turkey; Samsun (Nicotiana tabacum) and Hasankeyf (Nicotiana rustica). Salt treatments caused significant reduction in length, leafing, dry, and fresh weights together with a significant decline in relative water contents. Pigment levels decreased together with alterations in chlorophyll a/b and chlorophyll/carotenoid ratios. Anthocyanin levels and proline amount in roots increased. Malondialdehyde (MDA) and conductivity results indicated 0.6M NaCl induced damage in roots and leaves of Samsun. Since ascorbate peroxidase (APX) activity was higher in comparison to catalase (CAT), and increased whereas CAT activity decreased in response to salt treatment, APX seems to be more important in tobacco. Leaves and roots of cv. Samsun seems to be more effected by salt than Hasankeyf, whereas roots of Hasankeyf seems to have better adaptive responses against salt stress.
217. N. V. Ivanov, N. Sierro, and M. C. Peitsch, The Tobacco Plant Genome. Springer Nature, 2020. Abstract
     This book describes the history of tobacco genomics, from its “discovery” by Europeans to next-generation omics approaches in plant science. The authors primarily focus on the allotetraploid common tobacco plant (N. tabacum); however, separate chapters are dedicated to closely related Nicotiana species, such as N. benthamiana and N. attenuata, for which substantial progress in omics data analysis has been already achieved. While genetic maps, transcriptomes, and physical maps of BAC libraries have significantly enhanced our understanding of the tobacco plant, the genome of tobacco and related Nicotiana species has opened a new era in modern tobacco research. This book addresses current and future industrial and research applications as well as central challenges in tobacco science, including diseases, low variability of cultivars, the genome’s large size, polyploidy, and gene duplication.
218. S. K. R. Jadhav, K. A. Patel, B. B. Dholakia, and B. M. Khan, “Structural Characterization of a Flavonoid Glycosyltransferase from Withania Somnifera,” Bioinformation, vol. 8, no. 19, pp. 943–949, Oct. 2012. doi: 10.6026/97320630008943. Abstract
     Medicinal plants are extensively utilized in traditional and herbal medicines, both in India and around the world due to the presence of diverse low molecular weight natural products such as flavonoids, alkaloids, terpenoids and sterols. Flavonoids which have health benefits for humans are the large class of phenylpropanoid-derived secondary metabolites and are mostly glycosylated by UDP-glycosyltransferases (UGTs). Although large numbers of different UGTs are known from higher plants, very few protein structures have been reported till now. In the present study, the three-dimensional model of flavonoid specific glycosyltransferases (WsFGT) from Withania somnifera was constructed based on the crystal structure of plant UGTs. The resulted model was assessed by various tools and the final refined model revealed GT-B type fold. Further, to understand the sugar donors and acceptors interactions with the active site of WsFGT, docking studies were performed. The amino acids from conserved PSPG box were interacted with sugar donor while His18, Asp110, Trp352 and Asn353 were important for catalytic function. This structural and docking information will be useful to understand the glycosylation mechanism of flavonoid glucosides.
219. C. A. Jaleel, “Antioxidant Profile Changes in Leaf and Root Tissues of ’Withania Somnifera’ Dunal,” Plant Omics, vol. 2, no. 4, pp. 163–168, 2009. doi: 10.3316/informit.090724918670196. Abstract
     An important herbal plant Withania somnifera was evaluated for its antioxidant potential changes in leaves and roots. The main antioxidant constituents Non-enzymatic (ascorbic acid, alpha-tocopherol and reduced glutathione) and enzymatic (superoxide dismutase, ascorbate peroxidase, catalase, peroxidase and polyphenol oxidase) antioxidants were estimated from both leaves and roots. The analyses were carried out in the field-collected samples. It was found that plant contain a significant quantity of non-enzymatic and enzymatic antioxidants in these organs. But, the quantity varies in both organs. The roots contain more amounts of these antioxidants than leaves. Here from this study, it can be concluded that, the root of Withania somnifera are good source of nonenzymatic and enzymatic antioxidant components.
220. C. A. Jaleel, R. Gopi, P. Manivannan, and R. Panneerselvam, “Exogenous Application of Triadimefon Affects the Antioxidant Defense System of Withania Somnifera Dunal,” Pesticide Biochemistry and Physiology, vol. 91, no. 3, pp. 170–174, Jul. 2008. doi: 10.1016/j.pestbp.2008.03.006. Abstract
     Triadimefon is a triazole derivative, which have plant growth regulator properties. However, the influential mechanism of triadimefon on medicinal plants like Withania somnifera is not much studied. In the present investigation, the effects triadimefon at 10mgL−1 on the germination, early seedling growth, photosynthetic pigments, non-enzymatic antioxidant contents and activities of antioxidant enzymes were studied in W. somnifera Dunal plants. The germination percentage was not much affected by treatments and early seedling growth was reduced in terms of shoot length and leaf area but root length got increased with a concomitant enhancement in chlorophyll contents. The non-enzymatic antioxidants like ascorbic acid, reduced glutathione and α-tocopherol were increased in all parts (root, stem and leaf) of the seedlings. Triadimefon treatment caused an increase in the activities of antioxidant enzymes like superoxide dismutase, peroxidase, polyphenol oxidase and catalase. From our results it can be concluded that, the triadimefon can be used as a potential tool to enhance the antioxidant potential in medicinal plant W. somnifera.
221. C. A. Jaleel and M. M. Azooz, “Exogenous Calcium Alters Pigment Composition, γ-Glutamyl Kinase and Proline Oxidase Activities in Salt-Stressed Withania Somnifera.,” Plant Omics, vol. 2, no. 2, pp. 85–90, 2009. https://www.cabdirect.org/cabdirect/abstract/20093203217. Abstract
     In the present study effect of sodium chloride and calcium chloride on the proline metabolism of Withania somnifera plants was studied. The plants were treated with 100 mM NaCl, 5 mM CaCl2, 100 mM NaCl with 5 mM CaCl2 solutions. Ground water was used for irrigation to control plants. Plants were harvested randomly on 30 and 50 days after sowing (DAS). NaCl and CaCl2 stressed plants showed...
222. C. A. Jaleel, “Non-Enzymatic Antioxidant Changes in Withania Somnifera with Varying Drought Stress Levels.,” American-Eurasian Journal of Scientific Research, vol. 4, no. 2, pp. 64–67, 2009. https://www.cabdirect.org/cabdirect/abstract/20093256613. Abstract
     A study was conducted in Withania somnifera plants in order to analyse the alterations in non-enzymatic antioxidant contents under different water stress regimes. Plants were grown with different water regimes like 10 days interval drought (DID), 15 DID and 20 DID water stress. The plants were uprooted randomly after DID induction for determining the effect of water deficit on non-enzymatic and...
223. W. O. James, “Biosynthesis of the Belladonna Alkaloids,” Nature, vol. 158, no. 4019, pp. 654–656, Nov. 1946. doi: 10.1038/158654a0. Abstract
     THE origin of the tropane (and other) alkaloids in the plants that produce them has not been the object if much interest in the past. This seems to have been due to the sterility of the teleological appear to which they were subjected. They have been variously dismissed as reserve products of singular ineffectiveness, as flotsam thrown up on the beach of metabolism, and even as that final resort of the guesser hard up for a guess, a mechanism of detoxication. They are, nevertheless, very interesting substances for phytochemical investigation on account of their great variety, their relation to the proteins and the soluble nitrogen compounds, their specificity and the mystery of their coming and going. They are rendered especially suitable for investigation at the present time by the wealth of knowledge concerning their organic chemistry, and by the relative ease of their manipulation when compared with the other complex nitrogen compounds in the plant. The study of alkaloid metabolism may be expected to yield much information valuable to the wider study of the plant’s nitrogen metabolism in general.
224. A. Javaid and S. Shafique, “Herbicidal Activity of Withania Somnifera and Datura Alba against Rumex Denatus,” Journal of Agricultural Research (Pakistan), 2009. https://apply.jar.punjab.gov.pk/upload/1383485422_107_Paper_No.5_of_47(1).pdf. Abstract
     A study was conducted at the Institute of Mycology and Plant Pathology, University of Punjab, Quaid-e-Azam Campus, Lahore, Pakistan during 2008. Herbicidal activity of aqueous extracts of two medicinal plants of family Solanaceae viz. Withania somnifera Dunal and Datura alba Nees was evaluated against Rumex dentatus L., one of the most problematic weeds of wheat in Pakistan. Aqueous extracts of root and shoot of both test medicinal plant species resulted in pronounced suppression in germination as well as seedling growth of target weed species. However, variability in herbicidal potential of different types of employed extracts was evident. Germination was comparatively less susceptible while root growth in R. dentatus was highly susceptible to all aqueous extracts. Application of aqueous extracts caused 68 percent reduction in germination, 62 percent in shoot length, 96 percent in root length and 68 percent in seedling biomass.
225. A. Javaid, S. Shafique, and S. Shafique, “Herbicidal Activity of Withania Somnifera against Phalaris Minor,” Natural Product Research, vol. 24, no. 15, pp. 1457–1468, Sep. 2010. doi: 10.1080/14786410903169292. Abstract
     Herbicidal activity of Withania somnifera (L.) Dunal. was studied against Phalaris minor Retz., one of the most problematic weeds of wheat in Pakistan. In laboratory bioassays the aqueous, methanol and n-hexane extracts of 5, 10 and 15% w/v (fresh weight basis) of the roots and shoots of W. somnifera were applied. Extracts in the different solvents exhibited markedly variable herbicidal activities against germination and seedling growth of the target weed species. The methanol extracts showed the highest toxicity. Different concentrations of methanol shoot and root extracts declined the germination of P. minor by 21–71%, its shoot length by 40–72%, its root length by 50–99% and the plant biomass by 32–83%. The aqueous extracts proved to be comparatively less toxic than the methanol extracts, where generally the highest concentration of 15% exhibited pronounced toxicity against the target weed species. There was up to 48, 51, 99 and 55% suppression of the weed’s germination, shoot length, root length and plant biomass, respectively, due to the 15% aqueous root and shoot extracts. Generally, the n-hexane extracts of both roots and shoots exhibited insignificant or stimulatory effects against weed shoot length and plant biomass. In a foliar spray bioassay, aqueous and methanol shoot extracts of 10% w/v (dry weight basis) concentration were sprayed on one- and two-week old pot grown P. minor seedlings. Two subsequent sprays were carried out at five day intervals each. The aqueous extract significantly reduced the shoot and root dry biomass of one-week old P. minor plants. In a residue incorporation bioassay, crushed shoots of W. somnifera were incorporated in the soil at 1, 2, … 5% w/w. Phalaris minor seeds were sown one week after residue incorporation and plants were harvested 45 days after sowing. The lower concentrations of 2 and 3% significantly reduced, while higher concentrations of 4 and 5% of residue incorporation completely arrested, the germination of P. minor. The present study concludes that both roots and shoots of W. somnifera contain herbicidal constituents against P. minor.
226. A. Jhankare et al., “Development of Resistant Lines against Leaf Blight Disease of Withania Somnifera (L.) Dunal. Caused by Alternaria Alternata through in Vitro Selection.,” Plant Cell Biotechnology and Molecular Biology, vol. 12, no. 1/4, pp. 21–30, 2011. https://www.cabdirect.org/cabdirect/abstract/20193133638. Abstract
     Disease tolerant/resistant Withania (Withania somnifera (L.) cv JA-20 and MWS-100) cell lines were selected against leaf blight disease caused by Alternaria alternata. For this purpose callus and cell suspension cultures derived from mature embryos and hypocotyl explants were exposed to purified toxic culture filtrate produced by the fungus supplemented with MS culture medium. Two selection...
227. J. L. Jinks, “The Analysis of Continuous Variation in a Diallel Cross of Nicotiana Rustica Varieties,” Genetics, vol. 39, no. 6, pp. 767–788, Nov. 1954. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1209688/.
228. J. L. Jinks and H. S. Pooni, “Determination of the Environmental Sensitivity of Selection Lines of Nicotiana Rustica by the Selection Environment,” Heredity, vol. 49, no. 3, pp. 291–294, Dec. 1982. doi: 10.1038/hdy.1982.102. Abstract
     Selection has been practised for high and low final height in a population of 81 highly inbred lines derived by single seed descent from a random sample of the F2 of the cross of varieties 1 and 5 of Nicotiana rustica in the poorest, best and average of 15 environments. The properties of the resulting selections show that the high selection made in the best environment has a greater environmental sensitivity than that made in the poorest environment while the low selection made in the best environment has a lower environmental sensitivity than that made in the poorest environment. The selections made in the average environment have intermediate environmental sensitivities. These selections confirm the rules laid down by Jinks and Connolly (1975) relating the environmental sensitivities of selections to the selection environment.
229. J. L. Jinks, P. D. S. Caligari, and N. R. Ingram, “Gene Transfer in Nicotiana Rustica Using Irradiated Pollen,” Nature, vol. 291, no. 5816, pp. 586–588, Jun. 1981. doi: 10.1038/291586a0. Abstract
     In spite of reports of widespread matromorphy—the occurrence of diploid plants whose genomes are exclusively maternal in origin—in Nicotiana1 and in other genera2,3, attempts to induce it in pure-breeding material of Nicotiana rustica have met with little success4,5. When heavily irradiated pollen from one variety is used as the inducer on a contrasting variety acting as maternal parent, the progeny, while showing a greater resemblance to the maternal parent, are not true matromorphs because they usually have at least one characteristic of the paternal parent. At the highest dose of γ-irradiation, 20 krad, the progeny usually acquire from the paternal parent only one or a small sample of the many heritable characteristics by which the two varieties differ. By selecting within these progenies we have shown here that lines can be isolated with the characteristics of the pure-breeding maternal variety but with the exception of a specific characteristic transferred from the paternal variety. To achieve the same end by conventional means would require a combination of many generations of recurrent backcrossing and selection.
230. J. L. Jinks and D. T. Coombs, “The Relationship between Major Gene Controlled Inflorescence Morphology and Continuous Variation for Final Height in Nicotiana Rustica,” Heredity, vol. 42, no. 3, pp. 299–307, Jun. 1979. doi: 10.1038/hdy.1979.33. Abstract
     Inflorescence morphology is a major discontinuity among Nicotiana rustica varieties and is controlled by major genes at, at least, two loci, which display duplicate non-allelic interactions. Its contribution to continuous variation for final height has been examined in F1, F2 and backcross families of a diallel set of crosses among eight inbred lines and in the F9 families of one of the crosses. It makes no direct contribution, but if all plants are divided into mop and non-mop types of inflorescence most of the variation in final height within each type can be accounted for by the variation in flowering time. Inflorescence morphology appears to switch the correlation between flowering time and final height from a very high negative value within the mops to a very high positive value within non-mops. Most of the non-allelic interactions for final height occur in crosses between a mop and a non-mop parent.
231. A. C. Jnanesha and A. Kumar, “Improved Production and Postharvest Technologies in Ashwagandha (Indian Ginseng),” in Medicinal Plants, Apple Academic Press, 2022. doi: 10.1201/9781003277408-8. Abstract
     Withania somnifera is a miracle medicinal crop used traditionally in Ayurveda and Siddha for treating various diseases in South Asia. The root is the principal productive part of the Ashwagandha plant employed widely in the Indian system of medicine. The root is having potential medicinal properties and contains several alkaloids. One of the major alkaloids, Withanoloides, helps improving reproductive system in men and women, antistress, antioxidant, antiaging, diuretic, hypothyroid, immunomodulatory, antidementia, antihyperglycemic, anti-hypercholesterolemic, and cardiovascular activity. Leaves contain Withaferin A and B used for the treatment of thyroid and insomnia problems. It is drought tolerant cultivated mainly in Andhra Pradesh, Telangana, Karnataka, Rajasthan, Madhya Pradesh, and other states of India. Simple cultivation practices and higher root price attracting the farmer to grow Withania somnifera crop in large scale. The main challenges posed by farmers are interference of middlemen, lack of organized market, fluctuation in root price, demand-supply of roots, climatic variation, lower root yield, lack of availability of improved varieties, high fiber content in some location, lack of knowledge about improved post-harvest technology and problem occurrence with long term storage of roots.
232. L. Johny, X. A. Conlan, A. Adholeya, and D. M. Cahill, “Growth Kinetics and Withanolide Production in Novel Transformed Roots of Withania Somnifera and Measurement of Their Antioxidant Potential Using Chemiluminescence,” Plant Cell, Tissue and Organ Culture (PCTOC), vol. 132, no. 3, pp. 479–495, Mar. 2018. doi: 10.1007/s11240-017-1344-y. Abstract
     Markedly increased withanolide content was found in transformed roots (TR) of Withania somnifera germplasm grown in low mineral minimal media and withanolides showed high antioxidant potential when analysed using acidic potassium permanganate chemiluminescence. Transformation frequency of explants infected with Agrobacterium rhizogenes strain A4 varied between the three germplasms tested with the highest observed as 75\,± 0.9. Transformed root production was explant specific with leaves being the most productive among the different explants used. Withanolides, namely withaferin A, withanolide A, withanolide B and 12-deoxywithastramonolide were detected in TR cultures and differences in their content were found between germplasms. The highest concentrations of secondary metabolites were found in 4-week-old cultures and concentrations declined by the 8th and 12th week of culture. In 4-week-old cultures, the biomass of TR cultures was 4.5 fold higher than their respective non-transformed roots (NTR). Withaferin A was found in TR at levels that were 28–34 times higher than that found in NTR. A rapid method for the determination of the antioxidant potential of W. somnifera TR extracts was developed using post-column acidic potassium permanganate chemiluminescence (APPC) detection. The APPC chromatographic peaks for extract constituents showed strong alignment with those found for ultraviolet absorbance detection. The methods developed in this study for TR culture establishment and the use of a fast and sensitive way for the qualitative and quantitative determination of the antioxidant activity of their metabolites provides a new platform that will have use for similar studies in other species.
233. C. Joshi, N. Gajbhiye, A. Phurailatpam, K. A. Geetha, and S. Maiti, “Comparative Morphometric, Physiological and Chemical Studies of Wild and Cultivated Plant Types of Withania Somnifera (Solanaceae),” Current Science, vol. 99, no. 5, pp. 644–650, 2010. https://www.jstor.org/stable/24108318. Abstract
     Ashwagandha (Withania somnifera Dunal.) is an important medicinal plant cultivated in about 4000 ha in India. It is mainly distributed in the northwestern region of Madhya Pradesh and also in limited areas of different states. However, wild collection of the species continues to be a source of raw drug in Ayurvedic preparations. In the present communication, results of a comparative study of the wild type distributed locally in Anand, Gujarat and the superior variety, Jawahar Ashwagandha (JA-20), are presented. Results revealed conspicuous differences between the cultivated and wild-type plants in most of the characters studied. One of the major differences between cultivated and wild-type plants is that the former are annual, whereas the latter are perennial. Photosynthetic rate was higher in the wild type, which was reflected in its higher biomass production. Another distinguishing character was the floral structure which favours self-pollination in the cultivated plants because of short stigma covered with anther lobes, which is in contrast to the wild type having long, projected stigma inviting cross-pollination. The cultivated plants were in full bloom during December–February; however, in the wild type flowering was a continuous process throughout its lifespan. Flow cytometer study revealed the same ploidy level for both the plant types. However, chemical profile showed variation between the two plant types, even though targeted chemical constituents tested in the study were common to both. However, HPLC quantification of these constituents showed superiority of the wild type compared to JA-20.
234. P. Joshi, L. Misra, A. A. Siddique, M. Srivastava, S. Kumar, and M. P. Darokar, “Epoxide Group Relationship with Cytotoxicity in Withanolide Derivatives from Withania Somnifera,” Steroids, vol. 79, pp. 19–27, Jan. 2014. doi: 10.1016/j.steroids.2013.10.008. Abstract
     Withania somnifera is one of the highly reputed medicinal plants of India. Its steroidal constituents exist in the form of two major substitution patterns, viz. withaferin A (1) and withanone (5). Withaferin A with oxidation at carbons 4, 5, and 6 is considered as an active type, especially as anticancer, whereas the withanones with oxidation at carbons 5, 6, and 7 rarely show any activity. We prepared a series of derivatives with modifications at carbons 5, 6, and 7 in ring B of these withanolides to study the role of the epoxide group towards the cytotoxic property of these bioactive steroids. We have converted withanolides into the respective thiiranes, amino alcohols and alcohols by selective reactions at the epoxide ring and were evaluated for in vitro anticancer activity against four cancer cell lines to study the structure activity relationships. The transformations of the epoxide group in withanolides of the withaferin A type showed moderate reduction in their cytotoxicity whereas the almost inactive withanones have shown some improvements in their alcohol derivatives.
235. N. R. Joshi, M. A. Patel, and K. N. P. and A. D. Patel, “Genetic Variability, Correlation and Path Analysis in Ashwagandha [Withania Somnifera (L.) Dunal],” Electronic Journal of Plant Breeding, vol. 5, no. 4, pp. 875–880, Dec. 2014. https://www.ejplantbreeding.org/index.php/EJPB/article/view/309. Abstract
     Forty diverse genotypes of Ashwagandha [Withania somnifera (L.)Dunal] were evaluated for 12 quantitative characters. Higher magnitude of genotypic and phenotypic coefficient of variation was recorded for leaf width, number of primary branches per plant, number of secondary branches per plant, root length, dry weight of root, withanoloide content and starch content. High heritability in conjuction with high genetic advance were observed for dry weight of root, root length, diameter of root at collar region, withanoloide content and starch content which indicated that selection could be effective for these traits. The estimates of correlation coefficient revealed that the genotypic correlations were higher than their corresponding phenotypic correlation for all characters. It was observed that root length, diameter of root at collar region, plant height, leaf length, leaf width, days to flower initiation and days to maturity showed significant positive association with dry weight of root at genotypic as well as phenotypic levels. Path analysis revealed that plant height, days to maturity, diameter of root at collar region and leaf width were major characters having positive direct effect and significant association with dry root yield per plant. Hence selection for these trait would be quite effective to improve dry root yield in Ashwagandha.
236. N. R. Joshi, M. A. Patel, K. N. Prajapati, and J. R. P. and A. D. Patel, “Genetic Diversity In Ashwagandha (Withania Somnifera (L.) Dunal),” Electronic Journal of Plant Breeding, vol. 6, no. 3, pp. 870–874, Nov. 2015. https://ejplantbreeding.org/index.php/EJPB/article/view/204. Abstract
     Genetic divergence among 40 ashwagandha accessions of different geographic origin was assessed using Mahalanobis D2 statistics. Observations revealed significant genotypic differences and accordingly genotypes were classified into six clusters. Cluster I was the largest with thirty genotypes followed by II and III clusters which have four and three genotypes, respectively. Cluster IV, V and VI contained only one most divergent genotype. The maximum inter-cluster distance (D=1538.09) was found between cluster II and VI, followed by that between II and III (D=983.03). The minimum inter-luster distance was observed between cluster Iand II (D=285.09). Theintra-­cluster distance (D) ranged from 142.22 (cluster-III) to 192.10 (cluster-I). The three clusters (IV, V and VI) contained single genotype each and therefore, their intra-cluster distances were zero. The genotype of cluster VI was unique as it was having highest values for leaf width, diameter of root at collar region with high dry root yield. The cluster II was desirable in respect of days to flower initiation and days to maturity and also had highest value for number of primary branches per plant, withanoloide content and starch content. The cluster III exhibited highest value for plant height and leaf length. The cluster IV had highest mean values for number of secondary branches per plant and root length. Thus, hybridization among these genotypes can generate desirable transgressive segregants.
237. A. G. Joshi and M. A. Padhya, “Shoot Regeneration from Leaf Explants of Withania Somnifera (L.) Dunal,” Notulae Scientia Biologicae, vol. 2, no. 1, pp. 63–65, Mar. 2010. doi: 10.15835/nsb213609. Abstract
     Regeneration from leaf explants of Withania somnifera (L.) for mass propagation was studied on Murashige and Skoog’s medium supplemented with Kinetin (Kn) and 6-benzylaminopurine (BAP) alone or in combination. Shoot buds were induced from the midrib on the abaxial side in presence of Kn and BAP (4 µM). These shoot buds developed into shoots on the same medium. Rooting of these shoots was achieved in 0.5 µM of IBA.
238. N. Joshi, “In Vitro Growth and Shoot Multiplication in Nicotiana Tabacum L. - Influence of Gelling Agent and Carbon Source,” International Journal of Plant Developmental Biology, vol. 3, no. 1, pp. 29–33, 2009. Abstract
     The present study examined the influence of various gelling agents and carbon sources on in vitro growth and multiplication in Nicotiana tabacum L. cv. ‘Havana 425’ (Family: Solanaceae). Shoot multiplication was greatly favoured on medium gelled with reduced concentrations of agar. Complete absence of agar in the medium evoked a better response than that obtained on agar-containing medium. Replacement of agar by guar gum (at 2.0%, w/v) showed a two-fold improvement in in vitro growth and multiplication. On this medium ca. 12 elongated shoots were obtained. On medium gelled with Phytagel (0.1%), the rate of shoot multiplication was two times higher than that recorded on control. Incorporation of different carbon sources in the range of 1.5-4.0% (w/v) evoked varied responses in terms of shoot growth and multiplication. Glucose at 4.0% was the most effective carbon source where a maximum number of elongated shoots (>1.6 cm in height) were produced. Wet and dry weights of such shoots were also highest. The results emphasized the potential of guar gum as a gelling agent and glucose as a carbon source for increasing shoot multiplication and growth of N. tabacum.
239. L. JuGou, K. HongMei, D. JinRan, S. WenLong, M. WenGuang, and C. SuiYun, “Effects of temperature on Nicotiana stocktonii during microsporogenesis.,” Guangxi Zhiwu / Guihaia, vol. 33, no. 5, pp. 614–619, 2013. https://www.cabdirect.org/cabdirect/abstract/20143009048. Abstract
     In order to explore the influence of temperature on the meiosis of pollen mother cells and microsporogenesis in Nicotiana stocktonii, materials planted at different temperatures were observed by carbol fuchsin staining. The results showed that there was less abnormality and lower microkernel ratio during meiosis of pollen mother cells and microsporogenesis when N. stocktonii was planted at higher...
240. M. Jullien, D. Bouinot, S. Ali-Rachedi, B. Sotta, and P. Grappin, “Abscisic Acid Control of Seed Dormancy Expression in Nicotiana Plumbaginifolia and Arabidopsis Thaliana.,” Dormancy in plants: from whole plant behaviour to cellular control, pp. 195–210, 2000. https://www.cabdirect.org/cabdirect/abstract/20000711436. Abstract
     The involvement of ABA in maintaining dormancy in imbibed seeds of Nicotiana plumbaginifolia and Arabidopsis thaliana was investigated. The germination behaviour of freshly harvested and after-ripened seeds was characterized. Using delay in germination as the criteria of dormancy expression, the effects of ABA, GA and fluridone on dormant and non-dormant imbibed seeds were determined. ABA content...
241. J. Kahl et al., “Herbivore-Induced Ethylene Suppresses a Direct Defense but Not a Putative Indirect Defense against an Adapted Herbivore,” Planta, vol. 210, no. 2, pp. 336–342, Jan. 2000. doi: 10.1007/PL00008142. Abstract
     Herbivory induces both direct and indirect defenses in plants; however, some combinations of these defenses may not be compatible. The jasmonate signal cascade activated both direct (nicotine accumulations) and indirect (mono- and sesquiterpene emissions) whole-plant defense responses in the native tobacco Nicotiana attenuata Torr. Ex Wats. Nicotine accumulations were proportional to the amount of leaf wounding and the resulting increases in jasmonic acid (JA) concentrations. However, when larvae of the nicotine-tolerant herbivore, Manduca sexta, fed on plants or their oral secretions were applied to leaf punctures, the normal wound response was dramatically altered, as evidenced by large (4- to 10-fold) increases in the release of (i) volatile terpenoids and (ii) ethylene, (iii) increased (4- to 30-fold) accumulations of endogenous JA pools, but (iv) decreased or unchanged nicotine accumulations. The ethylene release, which was insensitive to inhibitors of induced JA accumulation, was sufficient to account for the attenuated nicotine response. Applications of ethylene and ethephon suppressed the induced nicotine response and pre-treatment of plants with a competitive inhibitor of ethylene receptors, 1-methylcyclopropene, restored the full nicotine response. This ethylene burst, however, did not inhibit the release of volatile terpenoids. Because parasitoids of Manduca larvae are sensitive to the dietary intake of nicotine by their hosts, this ethylene-mediated switching from direct to a putative indirect defense may represent an adaptive tailoring of a plant’s defense response.
242. V. S. Kandalkar, H. Patidar, and K. B. Nigam, “Genotypic Association and Path Coefficient Analysis in Ashwagandha (Withania Somnifera),” INDIAN JOURNAL OF GENETICS AND PLANT BREEDING, vol. 53, no. 03, pp. 257–260, Oct. 1993. https://www.isgpb.org/journal/index.php/IJGPB/article/view/2876. Abstract
     Genotypic and phenotypic correlations and path coefficient for seven characters were studied in 36 early generation selections in F2 population of wide crosses (WS 20 X Wild and WS 22 XWild) and seven genotypes of ashwagandha. Results indicated that root yield was significantly and positively correlated with plant height, stem branches, root length, root diameter, root branches, and seed yield. All the component characters studied were significantly and positively associated among themselves, except root branches with seed yield. Path coefficient analysis showed highest positive direct and indirect effect of plant heightand stem branches on root yield. Indirect effects ofothercomponent characters were high through plant height and stem branches on root yield. Selection for high plant-height and stem branches appeared to be useful for improving root yield.
243. F. E. Kandil, N. H. El Sayed, A. M. Abou-Douh, M. S. Ishak, and T. J. Mabry, “Flavonol Glycosides and Phenolics from Withania Somnifera,” Phytochemistry, vol. 37, no. 4, pp. 1215–1216, Nov. 1994. doi: 10.1016/S0031-9422(00)89563-1. Abstract
     6,8-Dihydroxykaempferol 3-rutinoside, a new compound, along with quercetin and its 3-O-rutinoside and 3-rutinoside-7-glucoside, were isolated from the leaves of Withania somnifera. In addition, three known aromatic acids were detected.
244. M. Kang, H. Ahn, E. Rothe, I. T. Baldwin, and S.-G. Kim, “A Robust Genome-Editing Method for Wild Plant Species Nicotiana Attenuata,” Plant Biotechnology Reports, vol. 14, no. 5, pp. 585–598, Oct. 2020. doi: 10.1007/s11816-020-00634-5. Abstract
     CRISPR genome-editing techniques theoretically enable us to edit any genes in any plants. However, plant tissue culture is required for generating targeted mutants in plants, except in some model plant species such as Arabidopsis thaliana. To modify ecologically or agronomically important traits in plants using the CRISPR system, a robust plant regeneration method and efficient genome-editing tools must be developed. This study shows the entire process of genome editing and the regeneration process for wild tobacco, Nicotiana attenuata. We delivered T-DNA harboring Streptococcus pyogenes Cas9 (SpCas9) and guide RNA (gRNA) via Agrobacterium-mediated transformation into the hypocotyl cells of the wild tobacco and regenerated gene-edited plants. The efficacy of genome editing was measured in N. attenuata protoplasts in which SpCas9 and gRNA were transiently expressed. Light intensity (476.66–627.00 μW/cm2, 20.69–52.21 μE) was optimized to enhance the emergence of plant shoots during callus induction, and the core step of dealing with plant tissues was recorded. In addition, we found that wounding the bottom part of mature plants was critical for root regeneration. By tracking mutation patterns and efficiency at each regeneration step, we found that the mutation was induced early on in the tissue culture process and maintained throughout the regeneration process. Genome-editing techniques have opened the way to study the function of the genes in all plants, and this study will provide guidelines for editing a gene in the plant of interest.
245. N. D. Kannan, G. Kul, and aivelu, “Drought Induced Changes in Physiological, Biochemical and Phytochemical Properties of Withania Somnifera Dun.,” Journal of Medicinal Plants Research, vol. 5, no. 16, pp. 3929–3935, Aug. 2011. doi: 10.5897/JMPR.9000462. Abstract
     Drought (D) stress effects on growth, photosynthetic pigments, photosynthetic electron transport, thylakoid polypeptides and phytochemical variations of Withania (Solanaceae), a tropical medicinal herb were studied. Exposure of plants to D-stress led to noticeable decrease in leaf area, photosynthetic pigments, root and shoot lengths and photosynthetic activity. Well-watered plants maintained high content of total chlorophyll (Chl), root and shoot lengths, leaf area and photosynthesis. We analysed alterations at cellular level of the D-stress-associated proteins at different time intervals. A set of proteins in the range of 34 to 40 kDa showed variations in response to D-stress. Qualitative high performance thin layer chromatography (HPTLC) analysis of root extract obtained from control and D-stressed plants showed quantitative and qualitative variations. Withaferin A content increased 5% under D-stress as compared to control. Key words: Carotenoids, chlorophyll, fluorescence, leaf area, pigments photosynthesis.
246. R. Kartusch and B. Mittendorfer, “Ultraviolet Radiation Increases Nicotine Production in Nicotiana Callus Cultures,” Journal of Plant Physiology, vol. 136, no. 1, pp. 110–114, Apr. 1990. doi: 10.1016/S0176-1617(11)81623-8. Abstract
     The effect of chronic UV-A and B irradiation with low intensity on growth and nicotine production was investigated in pith tissue callus cultures of Nicotiana tabacum L. Growth was not influenced by visible light, UV-A or UV-B under the irradiation conditions used. Nicotine production was suppressed by visible light. Within a certain dose range, UV-A stimulated nicotine production. UV-B showed no effect.
247. P. J. Katba, N. D. Dholariya, and A. D. Patel, “Genetic Variability among Different Characters in Tobacco (Nicotiana Rustica L.).,” Trends in Biosciences, vol. 7, no. 22, pp. 3691–3694, 2014. Abstract
     Fourty genotypes of tobacco (Nicotiana rustica L.) studied for variability parameters during 2012-13. The analysis of variance revealed significant differences for all the characters studied. The genetic variance contributed major proportion of total variance for all characters suggesting that these characters are under genetic control. The high genotypic and phenotypic coefficient of variation...
248. P. J. Katba, B. L. Pandor, and L. N. Gediya, “Genetic Association among Different Characters in Tobacco (Nicotiana Rustica L.),” Journal of Pharmacognosy and Phytochemistry, vol. 8, no. 1, pp. 146–148, 2019. https://www.phytojournal.com/archives/2019.v8.i1.6714/genetic-association-among-different-characters-in-tobacco-ltemgtnicotiana-rustica-ltemgtl. Abstract
     Forty genotypes of tobacco (Nicotiana rustica L.) studied for correlations and path analysis during 2012-13. The estimates of correlation coefficient revealed that cured leaf yield was positively correlated at both genotypic and phenotypic levels with days to flowering, number of leaves per plant, plant height, days to maturity, leaf length and leaf width, while it was negatively correlated with leaf thickness and reducing sugar content. Path analysis based on genotypic correlation showed that number of leaves per plant, plant height and leaf length are important characters that exerted considerable direct effect on cured leaf yield revealing scope for considering these characters in selection programme for bringing out desired improvement in tobacco yield.
249. P. J. Katba, R. G. Hadiya, V. N. Kapadia, D. C. Patel, and A. D. Patel, “Genetics Studies on Yield and Pharmaceutical Quality Parameters in Tobacco (Nicotiana Rustica L.),” Journal of Pharmacognosy and Phytochemistry, vol. 6, no. 1, pp. 399–404, 2017. https://www.phytojournal.com/archives/2017.v6.i1.1106/genetics-studies-on-yield-and-pharmaceutical-quality-parameters-in-tobacco-nicotiana-rustica-l. Abstract
     Tobacco is one of the world’s creditable crops. This relevant study significant and desirable determine of variability due variance of specific combining ability (sca) suspend significantly for all the traits under study indicating the importance of non-additive gene action for the inheritance of these characters. As variance due to both GCA and SCA were found significant for most of the character appeared to be under the influence of both additive and non-additive gene actions. The estimates of gca effects suggested those parents, GC 1, AR 72, Motihari Hemti and Sel.15-16 were good general combiners for cured leaf yield and its related attributes. The estimates of sca effects indicated that the cross GC 1, AR 72, Motihari Hemti and Sel.15-16 were the most promising for cured leaf yield and some of its related traits. The success of any breeding programme largely depends on choice of parents and breeding procedure adopted. Persistent efforts are being made to improve yield and yield contributing characters in tobacco through hybridization.
250. K. Kattimani, Y. N. Reddy, and B. R. Rajeswara Rao, “Effect of Pre-Sowing Seed Treatment on Germination, Seedling Emergence, Seedling Vigour and Root Yield of Ashwagandha (Withania Somnifera Dunal),” Seed Science and Technology, vol. 27, pp. 483–488, Jan. 1999. Abstract
     A field experiment was conducted at the College of Ag ricultule, Rajendranagar, Hyderabad to study the effect of pre-sowing seed treatments on seed vigour, root length and dry root yield of Ashwagandha under the agro-climatic conditions of Andhra Pradesh. Seedling vigour revealed that seeds soaked with nitrates of sodium and potassium at 1.0% for 24 h produced more vigourous seedlings, higher dry matter accumulation and root length as compared to unsoaked and water soaked seeds. Unsoaked seeds recorded minimum root length throughout the different stages of crop growth except l5 days after sowing. Seeds soaked with nitrate of sodium and potassium at 1.0% recorded maximum dry root yield of 3.93 and 3.43 g/plant respectively.
251. K. N. Kattimani, Y. N. Reddy, and B. R. Rajeswara Rao, “Influence of Pre-Sowing Seed Treatments on Seedling Vigour, Root Length and Dry Root Yield of Ashwagandha Withania Somnifera under Semi-Arid Tropical Climate of Hyderabad.,” Journal of Medicinal and Aromatic Plant Sciences, vol. 22 & 23, pp. 221–223, Jan. 2001. Abstract
     A field study was undertaken on the response of pre-sowing seed treatments with nitrates of potassium, cobalt, ammonium, sodium, calcium and zinc for 24h and their impact on root length and dry root yield of ashwagandha var. Jawahar 20. Pre-soaking the seeds with nitrates of sodium, potassium and calcium showed beneficial effects on root length and dry root yield under the semi-arid tropical conditions of Hyderabad.
252. S. Kaul et al., “Novel Methods to Generate Active Ingredients-Enriched Ashwagandha Leaves and Extracts,” PLOS ONE, vol. 11, p. e0166945, Dec. 2016. doi: 10.1371/journal.pone.0166945. Abstract
     Ashwagandha (Withania somnifera) is an Ayurvedic herb commonly used in world-renowned traditional Indian home medicine system. Roots of Ashwagandha have been traditionally known to possess a variety of therapeutic and health promoting potentials that have not been sufficiently supported by laboratory studies. Nevertheless, most, if not all, of the preventive and therapeutic potentials have been assigned to its bioactive components, steroidal alkaloids and lactones. In contrast to the traditional use of roots, we have been exploring bioactivities in leaves of Ashwagandha. Here, we report that the leaves possess higher content of active Withanolides, Withaferin-A (Wi-A) and Withanone (Wi-N), as compared to the roots. We also established, for the first time, hydroponic cultivation of Ashwagandha and investigated the effect of various cultivation conditions on the content of Wi-A and Wi-N by chemical analysis and bioassays. We report that the Withanone/Withaferin A-rich leaves could be obtained by manipulating light condition during hydroponic cultivation. Furthermore, we recruited cyclodextrins to prepare extracts with desired ratio of Wi-N and Wi-A. Hydroponically grown Ashwagandha and its extracts with high ratio of withanolides are valuable for cancer treatment.
253. M. K. Kaul, A. Kumar, A. Ahuja, B. A. Mir, K. A. Suri, and G. N. Qazi, “Production Dynamics of Withaferin A in Withania Somnifera (L.) Dunal Complex,” Natural Product Research, vol. 23, no. 14, pp. 1304–1311, Sep. 2009. doi: 10.1080/14786410802547440. Abstract
     Withaferin A (WA) is an important withanolide holding promise in cancer treatment and as a relatively safe radiosensitive/chemotherapeutic agent, which is present in traces in all parts of Withania somnifera except the leaves, where as it is reported to be present in only two non-Indian chemotypes (South African chemotype/Israel chemotype 1). The present studies have marked its presence in all Indian populations (wild/cultivated), as well as two identified Indian chemotypes (AGB002 and AGB025). The quantitative dynamics of WA production in Indian populations and interchemotypic hybrids developed at our institute have been studied, and the results were compared with five previously reported chemotypes from Israel, South Africa and India. An analysis on inheritance characteristics based on presence/absence of WA in hybrid plants and their respective parents is given for future studies on the chemogenetics of this complex species in greater detail. Further, the production potential of WA in vitro propagated plants of elite varieties developed at our institute is discussed, in view of maintaining chemotypic fidelity and stability from a production point of view. Also, evidence-based clues suggesting the leaves as the site of the synthesis of WA is provided.
254. M. K. Kaul, A. Kumar, and A. Sharma, “Reproductive Biology of Withania Somnifera (L.) Dunal,” Current Science, vol. 88, no. 9, pp. 1375–1377, 2005. https://www.jstor.org/stable/24110700.
255. K. Kaur et al., “Biotechnological Approaches in Propagation and Improvement of Withania Somnifera (L.) Dunal,” in Science of Ashwagandha: Preventive and Therapeutic Potentials, S. C. Kaul and R. Wadhwa, Eds. Cham: Springer International Publishing, 2017, pp. 459–478. doi: 10.1007/978-3-319-59192-6_22. Abstract
     Withania somnifera is a high value medicinal plant of the family Solanaceae. It is known for its anti-tumour, anti-stress, anti-aging, cardio-protective, neuro-protective and anti-inflammatory properties. Its pharmaceutical properties are attributed to a wide range of secondary metabolites, such as steroidal lactones, alkaloids, glycowithanolides, flavanol glycosides, phenolics and sterols. The traditional cultivation of W. somnifera is limited mainly by poor seed viability and germination, low yield and inconsistency in production of secondary metabolites. The infestation with various pests and pathogens also throw a major challenge in its cultivation. Biotechnological approaches involving organ, tissue and cell culture offer potential solution to the existing problems. In vitro propagation helps in rapid multiplication of elite cultivars and facilitate in raising quality planting materials. Genetic manipulation and secondary metabolite engineering hold great promise for enhancement of secondary metabolites and for overall crop improvement. The present chapter briefly discuss the challenges in W. somnifera and present a quick overview of biotechnological advances to address these challenges. It also highlights the futuristic approaches that would lay a foundation in the conglomeration of W. somnifera as an ideal model medicinal plant.
256. A. Kaur, A. Kaur, and P. Ohri, “Combined Effects of Vermicompost and Vermicompost Leachate on the Early Growth of Meloidogyne Incognita Stressed Withania Somnifera (L.) Dunal,” Environmental Science and Pollution Research, vol. 29, no. 34, pp. 51686–51702, Jul. 2022. doi: 10.1007/s11356-022-19264-1. Abstract
     Roots of Withania somnifera, an important medicinal herb, are prone to the infection of Meloidogyne incognita (a root parasitic nematode). The infection lowers the quality and quantity of plant material and poses a challenge in crop cultivation and obtaining desirable yield. In the present study, in vitro inhibitory activity of vermicompost leachate (Vcom-L) was assessed against % hatch and survival of M. incognita in a 96 h assay. Then, Vcom-L was used as soil supplement in combination with vermicompost (Vcom) to evaluate their nematode inhibitory and stress alleviating effect in W. somnifera, in a pot experiment. Root galling intensity and growth performance of nematode-stressed W. somnifera raised from seeds pre-soaked in distilled water (DW), Vcom-L, vermicompost tea (Vcom-T) and different dilutions of Vcom-L were assessed. We observed 79% suppression of egg hatching and 89% juvenile (J2) mortality after 96 h compared to control, at 100% concentration of Vcom-L. Significant reduction in gall formation with increase in growth parameters of seedlings was observed after combined application of Vcom (60% or 100%)\,+ Vcom-L and was evident as enhancement in seedling biomass and contents of chlorophyll and protein. However, proline, total phenolics and malondialdehyde (MDA) content declined significantly in these combinations compared to the control (0% Vcom). Activity of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidise (APX), guaiacol peroxidise (GPX) and glutathione reductase (GR) declined with Vcom as well as Vcom\,+ Vcom-L and corresponded with decline in the accumulation of reactive oxygen species in leaves. Further, 1:5 and 1:10 dilutions of Vcom-L in combination with Vcom (60%) proved beneficial in mitigating the nematode-induced stress in W. somnifera. Present results showed the potential of Vcom and Vcom-L in standardised combination as an effective strategy in controlling the pathogenicity of M. incognita in medicinal plants such as W. somnifera.
257. K. Kaur, D. Dolker, S. Behera, and P. K. Pati, “Critical Factors Influencing in Vitro Propagation and Modulation of Important Secondary Metabolites in Withania Somnifera (L.) Dunal,” Plant Cell, Tissue and Organ Culture (PCTOC), vol. 149, no. 1, pp. 41–60, May 2022. doi: 10.1007/s11240-021-02225-w. Abstract
     Withania somnifera (L.) Dunal is a valuable medicinal plant in the Solanaceae family. It is commonly known as Ashwagandha and is widely distributed around the globe. It has multiple pharmacological properties owing to the existence of diverse secondary metabolites viz., withanolide A, withanolide D, withaferin A, and withanone. It is in great demand in the herbal industry because of its extensive use. In this background, the major challenge lies in the rapid multiplication of elite cultivars of W. somnifera in order to produce genetically and phytoconstituents uniform plant material for pharmaceutical industries. Thus it is necessary to explore various biotechnological approaches for the clonal mass propagation and synthesis of pharmaceutically important constituents in W. somnifera. Though there are several studies on in vitro propagation on W. somnifera, yet many factors that critically influence the in vitro response and withanolides production need to be fine-tuned in the pretext of the existing knowledge. The current review focuses on the advancements and prospects in biotechnological interventions to meet the worldwide demands for W. somnifera and its bioactive compounds. This update on in vitro studies on W. somnifera will be useful to many researchers, entrepreneurs, and herbal industries looking for its in vitro mass multiplication and scientific utilization.
258. K. Kaur, P. Singh, K. Kaur, A. Bhandawat, P. Nogia, and P. K. Pati, “Development of Robust in Vitro Culture Protocol for the Propagation of Genetically and Phytochemically Stable Plants of Withania Somnifera (L.) Dunal (Ashwagandha),” Industrial Crops and Products, vol. 166, p. 113428, Aug. 2021. doi: 10.1016/j.indcrop.2021.113428. Abstract
     Withania somnifera (L.) Dunal is an important medicinal plant that has gained a lot of attention since ancient times due to its immense pharmaceutical potential. Scientific validations of its therapeutic potential against many diseases have immensely increased its demand in the domestic and international market. Thus, to narrow down the huge gap between its demand and supply, alternative strategies to conventional cultivation are required. In vitro propagation system provides a feasible option for the production of large-scale healthy, genetically and phytochemically uniform W. somnifera plants to boost the herbal industry and facilitate pre-clinical studies. However, the major challenge in in vitro propagation of W. somnifera is to maintain a high rate of shoot multiplication for prolonged periods in cytokinin containing MS medium. In the current investigation, for prolonged maintenance of in vitro shoots, a multi-step protocol involving the rotational subculture of Withania shoots in the auxin-cytokinin containing culture medium has been developed to minimize the adverse effects of cytokinin. In vitro shoots after prolonged subculture were checked for genetic uniformity and stable pharmaceutically important constituents. Furthermore, in vitro rooting was initiated either in an auxin-dependent or auxin-independent manner through modulation of ammonium nitrate strength. Hence, the protocol developed for in vitro propagation of W. somnifera is cost-effective and can be efficiently used for the industrial production of withanolides.
259. A. Kaur, P. Ohri, and D. A. Kaur, “Effect of Vermicompost Extracts on In-Vitro Germination and Growth of Withania Somnifera (L.) Dunal,” International Journal of Herbal Medicine, vol. 6, no. 2, pp. 28–32, 2018. https://www.florajournal.com/archives/2018/vol6issue2/PartA/7-1-1-510.pdf. Abstract
     Seed germination in Withania somnifera is low and erratic under natural conditions therefore in the present study we pre-soaked the seeds of W. somnifera for 12h in Vermicompost leachate (aerated and non-aerated), Vermicompost tea (aerated and non-aerated) and distilled water (DW). Treated seeds were germinated in vitro on filter papers moistened with respective pre-soaking treatments. Results indicated that seeds treated with Vcom-Ta resulted in highest germination (70%) and biomass/seedling (0.42g). Seed germination and biomass/seedling in different treatments was Vcom-Ta > Vcom-Tna > Vcom-Lna > Vcom-La > DW. Carbon content was significantly increased in Vcom-Lna, Vcom-La and Vcom-Ta treatments as compared to control. Nitrogen content was maximum in Vcom-Ta seedlings but there was a non significant difference between control, Vcom-Ta and Vcom-Lna. We observed more root hair proliferation in Vcom-Lna and Vcom-Ta treated seedlings. Therefore, the use of such organic treatments can be extended to cultivate W. somnifera under natural conditions.
260. A. Kaur, P. K. Pati, P. Ohri, and A. Kaur, “Effects of Vermicompost and Vermicompost Leachate on the Biochemical and Physiological Response of Withania Somnifera (L.) Dunal,” Journal of Soil Science and Plant Nutrition, vol. 22, no. 3, pp. 3228–3242, Sep. 2022. doi: 10.1007/s42729-022-00880-2. Abstract
     Withania somnifera is a widely used herb due to its multi-health benefits. There is a growing interest to improve the growth and chemical profile of this plant because a large gap exists in the supply and demand of the quality raw material. The present study focused on using organic amendments to enhance the health and quality of this herb.
261. M. Kaur, S. Sharma, and A. Mishra, “Influence of Phosphate Solubilizing Pseudomonas and Bacillus Strains on the Growth of Ashvagandha (Withania Somnifera),” Indian Journal of Agricultural Research, vol. 45, no. 2, pp. 128–133, 2011. https://arccjournals.com/journal/indian-journal-of-agricultural-research/. Abstract
     Twenty seven bacterial strains of both wenty seven bacterial strains of both wenty seven bacterial strains of both wenty seven bacterial strains of both wenty seven bacterial strains of both P PP PPseudomonas seudomonas seudomonas seudomonas seudomonas and and and and and Bacillus Bacillus Bacillus Bacillus Bacillus sp. from the rhizospher sp. from the rhizospher sp. from the rhizospher sp. from the rhizospher sp. from the rhizosphere ee ee of Ashvagandha were isolated and screened out for phosphate solubilization and production of of Ashvagandha were isolated and screened out for phosphate solubilization and production of of Ashvagandha were isolated and screened out for phosphate solubilization and production of of Ashvagandha were isolated and screened out for phosphate solubilization and production of of Ashvagandha were isolated and screened out for phosphate solubilization and production of other types of plant growth promoting activities affecting dir other types of plant growth promoting activities affecting dir other types of plant growth promoting activities affecting dir other types of plant growth promoting activities affecting dir other types of plant growth promoting activities affecting directly or indir ectly or indir ectly or indir ectly or indir ectly or indirectly ectly ectly ectly ectly. Out of twenty . Out of twenty . Out of twenty . Out of twenty . Out of twenty seven strains six strains , three of seven strains six strains , three of seven strains six strains , three of seven strains six strains , three of seven strains six strains , three of Bacillus Bacillus Bacillus Bacillus Bacillus sp. (WsNB-2, WsNB-6, and WsNB-13) and three of sp. (WsNB-2, WsNB-6, and WsNB-13) and three of sp. (WsNB-2, WsNB-6, and WsNB-13) and three of sp. (WsNB-2, WsNB-6, and WsNB-13) and three of sp. (WsNB-2, WsNB-6, and WsNB-13) and three of P PP PPseudomonas seudomonas seudomonas seudomonas seudomonas sp. (WsCP sp. (WsCP sp. (WsCP sp. (WsCP sp. (WsCP-1, WsCP -1, WsCP -1, WsCP -1, WsCP -1, WsCP-5 and WsCP -5 and WsCP -5 and WsCP -5 and WsCP -5 and WsCP-6) wer -6) wer -6) wer -6) wer -6) were selected to study the effect of phosphate e selected to study the effect of phosphate e selected to study the effect of phosphate e selected to study the effect of phosphate e selected to study the effect of phosphate solubilizing strains on the growth of rooted cutting of Ashvagandha and on other soil parameters solubilizing strains on the growth of rooted cutting of Ashvagandha and on other soil parameters solubilizing strains on the growth of rooted cutting of Ashvagandha and on other soil parameters solubilizing strains on the growth of rooted cutting of Ashvagandha and on other soil parameters solubilizing strains on the growth of rooted cutting of Ashvagandha and on other soil parameters like root colonization and phosphorous content of soil before and after the experiment. The like root colonization and phosphorous content of soil before and after the experiment. The like root colonization and phosphorous content of soil before and after the experiment. The like root colonization and phosphorous content of soil before and after the experiment. The like root colonization and phosphorous content of soil before and after the experiment. The selected strains significantly enhanced plant height after two months of plantation. Rhizosphere selected strains significantly enhanced plant height after two months of plantation. Rhizosphere selected strains significantly enhanced plant height after two months of plantation. Rhizosphere selected strains significantly enhanced plant height after two months of plantation. Rhizosphere selected strains significantly enhanced plant height after two months of plantation. Rhizosphere bacterial population incr bacterial population incr bacterial population incr bacterial population incr bacterial population increased significantly eased significantly eased significantly eased significantly eased significantly, which has r , which has r , which has r , which has r , which has resulted in nutrient mobilization and esulted in nutrient mobilization and esulted in nutrient mobilization and esulted in nutrient mobilization and esulted in nutrient mobilization and ultimately resulted in enhanced plant growth.
262. Y. Kaya and F. Z. Huyop, “An Easy and Reliable Method for Establishment and Maintenance of Tissue Cultures of Nicotiana Tabacum Cv TAPM 26,” International Journal of Science Letters, vol. 2, no. 2, pp. 62–71, Aug. 2020. doi: 10.38058/ijsl.764947.
263. Y. Kaya, “In Vitro Plant Regeneration of Tobacco (Nicotiana Tabaccum TAPM 26),” PhD thesis, UNIVERSITI TEKNOLOGI MALAYSIA, 2010. Abstract
     In the present research, a reproducible procedure has been developed for efficient plant regeneration of shoot from cotyledon explants of Nicotiana tabaccum TAPM 26. In vitro regeneration of Nicotiana tabaccum TAPM 26 was efficiently achieved applying combination of growth hormones supplemented into MS medium using cotyledon from 5-day-old and 7-day-old as explants. The growth hormones used were IAA for auxin (0.0, 0.1, 1, 10 mgL -1 ) and BAP for cytokinin (0.0, 0.1, 1, 10 mgL -1 ). The effects of growth hormones combination were assessed based on number of shoot and roots formation. Maximum number of shoots produced per explant was found when media continued with 10 µM BAP +0.0 µM IAA after 4 weeks in culture. Maximum number of root produced per explant was reached when media continued 10 µM IAA after 4 weeks in MS media. The current techniques proved that Nicotiana tabaccum was successfully propagated by direct organogenesis and can be applied for further research in making as transgenic Nicotiana tabaccum.
264. M. F. Keogh and D. G. O’Donovan, “Biosynthesis of Some Alkaloids of Punica Granatum and Withania Somnifera,” Journal of the Chemical Society C: Organic, no. 13, pp. 1792–1797, Jan. 1970. doi: 10.1039/J39700001792. Abstract
     The biosynthesis of isopeletierine [1-(2-piperidyl)propan-2-one], N-methylisopelletierine, and pseudopelletierine (9-methyl-9-azabicylo[3,3,1]nonan-3-one) in intact Punica granatum plants has been studied. N-Methylisopelletierine is shown to be derived from lysine and acetate. Acetate is shown to be the precursor of the three-carbon unit in isopelletierine and pseudopelletierine. N-Methylisopelletierine is shown to be the immediate precursor of psueudopelletierine.The derivation of anaferine from lysine and acetate has been demonstrated in intact Withania somnifera plants. Isopelletierine has also been shown to be a precursor of anaferine.
265. R. K. Khajuria et al., “Separation, Identification, and Quantification of Selected Withanolides in Plant Extracts of Withania Somnifera by HPLC-UV(DAD) – Positive Ion Electrospray Ionisation–Mass Spectrometry,” Journal of Separation Science, vol. 27, no. 7-8, pp. 541–546, 2004. doi: 10.1002/jssc.200301690. Abstract
     This paper describes a method for separation, identification, and quantification of selected withanolides in Withania somnifera plant extracts by HPLC-UV(DAD)–Mass Spectrometry (HPLC-MS). Withaferin-A (WS-3), 12-deoxywithastramonolide (WS-12DS), Withanolide A (WS-1), and Withanone (WS-2) were used as external standards. The compounds were isolated from Withania somnifera by repeated column chromatography of the root extract and their identity was established by 1H- and 13C-NMR and mass spectral data. The compounds were chromatographed on a Merck (250×4.6 mm ID, 5 μm) column and analyzed by Electrospray Ionization on a mass spectrometer in Selected Ion Mode (SIM). For quantification, [M + Na]+ ions were monitored. Linear calibration curves were obtained in the concentration range of 1.50 μg/mL to 6.5 μg/mL. The method was applied successfully to the detection and quantification of the said withanolides in a number of samples.
266. R. Khan, S. Shahzad, M. Choudhary, S. Khan, and A. Ahmad, “Communities of Endophytic Fungi in Medicinal Plant Withania Somnifera,” Pak. J. Bot, vol. 42, pp. 1281–1287, Apr. 2010. Abstract
     Withania somnifera (L.) Dunal is known to possess medicinal properties. Medicinal plants harbour endophytic mycoflora. Only a few plants have been studied for their endophyte biodiversity and their potential to produce bioactive secondary metabolites. There is a need to understand the biodiversity of endophytic fungi and their potential of producing novel compounds of medicinal importance. A total of 643 segments (202 leaf, 391 stem, and 50 root samples) from 20 different plants were screened for their endophytic mycoflora. Thirty-three fungal strains of 24 species have been isolated, four belonged to the class Ascomycetes and 20 to class Deuteromycetes. The highest species richness as well as frequency of colonization was in stem; with the exception of Aspergillus niger, A. terreus and A. alternata, all the other fungi were found to be organ-specific. In this study most dominant endophyte was found to be A. alternata. Overall colonization frequency was measured as 14.15%. Many of the pharmaceutical compounds produced by medicinal plants are reportedly produced by their endophytic fungi. Hence, it is important to study medicinal plants for their endophytic mycoflora for biodiversity and then to determine their medicinal properties. The present work was therefore initiated to study the endophytic fungal population in Withania somnifera (L.) Dunal., a commonly used medicinal plant in the subcontinent.
267. J. A. Khan, P. Srivastava, and S. K. Singh, “Sensitive Detection of a Phytoplasma Associated with Little Leaf Symptoms in Withania Somnifera,” European Journal of Plant Pathology, vol. 115, no. 4, pp. 401–408, Aug. 2006. doi: 10.1007/s10658-006-9029-0. Abstract
     Withania somnifera is an important medicinal plant native to the Indian-sub continent. Owing to the presence of a number of precious alkaloids, flavonoids and withanolides, it is widely used in the Indian and African systems of medicines. It is severely affected by phytoplasma present in the sieve tubes of phloem. With a view to micropropagate phytoplasma-free W. somnifera plants, an efficient and effective nested PCR-based system was developed for detection of associated phytoplasmas. Universal primers, designed from the 16S rDNA sequences of phytoplasmas, were applied in direct/nested-PCR. Total DNA extracts from leaf tissues of 33 suspected symptomatic and 11 non-symptomatic plants were subjected to direct PCR. The direct PCR products were subsequently employed as templates in nested PCR. The nested PCR could reamplify direct PCR products yielding a DNA fragment of 1.4 kb. A phytoplasma was detected in all the diseased plants and not from the healthy looking plants. Further, it was sensitive enough to amplify phytoplasma DNA obtained from crude DNA diluted up to 2500 times from naturally infected plants and also from various stages of in vitro-propagated diseased plants. Identical restriction fragment polymorphism enzyme profiles were obtained following restriction enzyme digestion of nested PCR products, obtained from five different plants, by EcoRI, AluI and RsaI restriction endonucleases. The developed nested PCR based system should facilitate indexing of the phytoplasma in different stages of in vitro-generated plants and probably identification of, as yet unknown, hosts and vectors of phytoplasma associated with phytoplasma disease of W. somnifera.
268. P. K. Khanna, A. Kumar, R. Chandra, and V. Verma, “Germination Behaviour of Seeds of Withania Somnifera (L.) Dunal: A High Value Medicinal Plant,” Physiology and Molecular Biology of Plants, vol. 19, no. 3, pp. 449–454, Jul. 2013. doi: 10.1007/s12298-013-0169-3. Abstract
     In order to evolve a quick method for smooth and optimum germination for Withania somnifera- a medicinally efficacious multipurpose plant, present investigation was carried to study the effect of physico-chemical treatments, storage, temperature, photoperiod and growth regulators (GA3, IAA, IBA, 2–4 D and BA) on germinability. The most effective treatment is GA3 at 150 μg/ml concentration at 25 °C. The optimal temperature for germination is 25 °C and continuous light favored germination showing that photoperiod has a significant role. The seedlings derived from seeds performed well when grown in a glasshouse. The data have implications for conservation and cultivation of the species studied.
269. S. Khatun, M. B. Ali, E.-J. Hahn, and K.-Y. Paek, “Copper Toxicity in Withania Somnifera: Growth and Antioxidant Enzymes Responses of in Vitro Grown Plants,” Environmental and Experimental Botany, vol. 64, no. 3, pp. 279–285, Dec. 2008. doi: 10.1016/j.envexpbot.2008.02.004. Abstract
     Indian ginseng (Withania somnifera L. Dunal), a medicinally important plant exposed to different concentrations of CuSO4 (0, 10, 25, 50, 100 and 200μM) for 30d in vitro accumulated high amount of Cu in a concentration manner. Toxic effects of Cu on plants were reflected by the reductions in fresh weight (FW), shoot and root length, chlorophyll and carotenoids contents. Malondialdehyde (MDA), a decomposition product of polyunsaturated fatty acids hydroperoxides, often considered as a suitable biomarker for lipid peroxidation was induced in metal-treated plants compared to the control. This hypothesis is also confirmed by the higher lipoxygenase (LOX) activity, coupled with higher hydrogen peroxide (H2O2) and superoxide anions (O2−) content in treated plants. Ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), glutathione-S-transferase (GST) and guaiacol-peroxidase (G-POD) activities of leaves were increased in the presence of Cu when compared to control plants indicated that antioxidant enzymes played an important role in protecting the plant from Cu toxicity. After native polyacrylamide gel electrophoresis (PAGE) analysis, six APX and four G-POD isoforms were detected and significantly induced in metal-treated plants as compared to control plants. The marked decrease in superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR) and glutathione peroxidase (GPx) activities seems to reflect its inability for eliminating the reactive oxygen species (ROS) resulting from Cu-induced oxidative stress. On the other hand, total phenolic contents increased with increasing concentration of Cu compared to the control. This study reveals that plants have the ability to grow in Cu polluted areas by altering various physiological changes. Since the level of total phenolic content in metal-treated plants increases, therefore, it is advisable to assess the biological activity of the plants before using for medicinal purposes.
270. C. T. Kiranoudis, Z. B. Maroulis, and D. Marinos-Kouris, “Mass Transfer Modeling for Virginia Tobacco Curing,” Drying Technology, vol. 8, no. 2, pp. 351–366, Jan. 1990. doi: 10.1080/07373939008959888. Abstract
     Four mathematical models, describing the moisture transfer during Virginia tobacco curing under varying air-temperature and humidity were tested using experimental data from an industrial dryer. The model, based on the assumption that the major resistance to moisture transfer is at the air boundary layer and that the driving force is the water activity difference, was found to be the most accurate. The appropriate mass transfer coefficient was determined. An empirical equation, predicting the equilibrium moisture content of Virginia tobacco as a function of temperature and water activity of the surrounding air, was proposed and experimentally tested.
271. I. Kirson, A. Abraham, and D. Lavie, “Chemical Analysis of Hybrids of Withania Somnifera L. (Dun.). 1. Chemotypes III (Israel) by Indian I (Delhi),” Israel Journal of Chemistry, vol. 16, no. 1, pp. 20–24, 1977. doi: 10.1002/ijch.197700007. Abstract
     Reciprocal crossbreeding between Withania somnifera chemotype III (Israel) and chemotype Indian I (Delhi) were performed. The F1 offspring of this cross was characterized by significant changes in the main steroidal lactone constituents of the parent plants. From the withaferin A (9) and the withanone (2) both present as major constituents in the parent Indian I, the former had disappeared in the hybrid and was replaced by withanolide D (10), whereas the latter was still present, though in about one third of the original quantity. It was accompanied by 17-deoxy-20-hydroxy withanone (2) and by withanolide T (1); the enzymatic system leading to hydroxylation at C20 (20αF-OH) has a dominant character. Two new withanolides have been isolated from this F1 offspring, namely withanolide T (1) (20αF-hydroxy withanone) and withanolide U (5) (4β-hydroxy-withanolide G). The structures assigned to these two new compounds are based on spectral evidence (NMR, IR and UV) and analysis of their fragmentation under electron impact.
272. I. Kirson, E. Glotter, A. Abraham, and D. Lavie, “Constituents of Withania Somnifera Dun—XI: The Structure of Three New Withanolides,” Tetrahedron, vol. 26, no. 9, pp. 2209–2219, Jan. 1970. doi: 10.1016/S0040-4020(01)92800-5. Abstract
     Three new withanolides (C28 steroids possessing a 6-membered ring lactone in the side chain) have been isolated from Withania somnifera (Solanaceae) growing in South Africa and characterized as 4β-hydroxy-1-oxo-5β,6β-epoxywitha-2,24-dienolide (IIIa), 4β,20α,(R)dihydroxy-1-oxo-5β,6β-epoxywith-2-enolide (IVa), and 4β-hydroxy-1-oxo-5β,6β-epoxywith-2-enolide (Va). The major steroidal components of the above plant are the known withaferin A (4β,27-dihydroxy-1-oxo-5β,6β-epoxywitha-2,24-dienolide) and withanolide D (4β,20α,(R)-dihydroxy-1-oxo-5β,6β-epoxywitha-2,24-dienolide). The structures assigned to compounds IIIa, IVa and Va are based on spectral evidence (NMR, IR and UV), analysis of their fragmentation under electron impact, as well as on chemical degradation to known compounds.
273. I. Kirson, E. Glotter, D. Lavie, and A. Abraham, “Constitutents of Withania Somnifera Dun. Part XII. The Withanolides of an Indian Chemotype,” Journal of the Chemical Society C: Organic, no. INVALID_SCITE_VALUE, pp. 2032–2044, Jan. 1971. doi: 10.1039/J39710002032. Abstract
     Eight new steroidal lactones of the withanolide series have been isolated from the leaves of Withania somnifera(Solanaceae) growing in North-Western India. This population of W. somnifera differs from those investigated so far and constitutes a new chemotype. The new compounds (IV)—(XI) have been investigated by chemical degradation and interconversion reactions, as well as by analysis of their spectral properties. The following structures have been assigned: 5α,17α-dihydroxy-1-oxo-6α,7α-epoxy-22R-witha-2,24-dienolide (IV); 5α,17α-dihydroxy-1-oxo-22R-witha-2,6,24-trienolide (V); 5α,27-dihydroxy-1-oxo-6α,7α-epoxy-22R-witha-2,24-dienolide (VIa); 4β,17α-dihydroxy-1-oxo-5β,6β-epoxy-22R-witha-2,24-dienolide (VIIa); 17α,27-dihydroxy-1-oxo-22R-witha-2,5,24-trienolide (VIIIa); 7α,27-dihydroxy-1-oxo-22R-witha-2,5,24-trienolide (VIIIa); 7α,27-dihydroxyl-1-oxo-22R-witha-2,5,24-trienolide (IXa); 1α,3β,5α-trihydroxy-6α,7α-epoxy-22R-with-24-enolide (Xa); 4β-hydroxy-5β,6β-epoxy-1-oxo-22R-witha-2,14,24-trienolide (XIa).
274. M. van Kleunen, M. Fischer, and S. D. Johnson, “Reproductive Assurance through Self-Fertilization Does Not Vary with Population Size in the Alien Invasive Plant Datura Stramonium,” Oikos, vol. 116, no. 8, pp. 1400–1412, 2007. doi: 10.1111/j.0030-1299.2007.16004.x. Abstract
     Autonomous self-fertilization is suggested to be associated with invasiveness in plants because it offers reproductive assurance when there is a shortage of suitable mates or pollinators. Given that shortages of mates and pollinators are a common cause of Allee effects in small plant populations, we predict that the benefits of self-fertilization in terms of reproductive assurance should be greatest in small populations. We tested this idea for the invasive herb Datura stramonium, a self-fertilizing species which is also cross-pollinated to some extent by insects (mainly hawkmoths and honeybees). During two consecutive years, we studied 20 and 55 populations, respectively, of different sizes. Untreated flowers of D. stramonium showed high levels of fruit and seed set in all populations studied. Although, fruit and seed set were generally reduced by about 90% in flowers in which self-fertilization was prevented through emasculation, this effect did not vary according to population size. By using a natural color (anthocyanin) dimorphism in 12 populations, we showed that the average outcrossing rate was low (1.3%) and that there was no relationship between outcrossing rate and population size. Pollen removal from flowers also did not vary according to population size, suggesting that the pollinator visitation rate is not lower in small populations. However, decreasing deviations of observed from expected fruit set with population size imply that small populations may have an increased chance of extinction due to demographic stochasticity. Overall, our results suggest that reproductive assurance through self-fertilization in invasive plants may be important for all stages of population establishment, and not just in the founder population.
275. K. L. Kohnen-Johannsen and O. Kayser, “Tropane Alkaloids: Chemistry, Pharmacology, Biosynthesis and Production,” Molecules, vol. 24, no. 4, p. 796, Feb. 2019. doi: 10.3390/molecules24040796. Abstract
     Tropane alkaloids (TA) are valuable secondary plant metabolites which are mostly found in high concentrations in the Solanaceae and Erythroxylaceae families. The TAs, which are characterized by their unique bicyclic tropane ring system, can be divided into three major groups: hyoscyamine and scopolamine, cocaine and calystegines. Although all TAs have the same basic structure, they differ immensely in their biological, chemical and pharmacological properties. Scopolamine, also known as hyoscine, has the largest legitimate market as a pharmacological agent due to its treatment of nausea, vomiting, motion sickness, as well as smooth muscle spasms while cocaine is the 2nd most frequently consumed illicit drug globally. This review provides a comprehensive overview of TAs, highlighting their structural diversity, use in pharmaceutical therapy from both historical and modern perspectives, natural biosynthesis in planta and emerging production possibilities using tissue culture and microbial biosynthesis of these compounds.
276. R. Kormelink, P. de Haan, D. Peters, and R. Y. R. 1992 Goldbach, “Viral RNA Synthesis in Tomato Spotted Wilt Virus-Infected Nicotiana Rustica Plants,” Journal of General Virology, vol. 73, no. 3, pp. 687–693. doi: 10.1099/0022-1317-73-3-687. Abstract
     The synthesis of viral RNA species in tomato spotted wilt virus-infected Nicotiana rustica plants was followed in terms of time and relative abundance. Systemic symptoms were visible after 4 days post-inoculation (p.i.), but viral (v) and viral-complementary (vc) strands of all three genomic RNA segments [large (L) RNA, medium (M) RNA and small (S) RNA] were detected from 2 days p.i. In addition, two subgenomic mRNAs, derived from S RNA, were detected. For the L RNA segment no subgenomic mRNAs were detected, suggesting that this segment is expressed via the synthesis of a genome-sized vc mRNA. A possible M-specific subgenomic mRNA was detected, showing a similar time course of appearance as the subgenomic mRNAs derived from the S RNA segment. Analysis of cytoplasmic RNA fractions revealed that both v and vc strands of all three genomic segments associate with the nucleocapsid protein into nucleocapsid structures, the vcRNA species being present in lower amounts. Intact, enveloped virus particles contained only the v strand of the L RNA segment and, surprisingly, both v and vc strands of the M and S RNA segment, though in different ratios.,
277. S. Kothari, C. Singh, Y. Kumar, and K. Singh, “Morphology, Yield and Quality of Ashwagandha (Withania Somnifera L. Dunal) Roots and Its Cultivation Economics as Influenced by Tillage Depth and Plant Population Density,” Journal of Horticultural Science, vol. 78, pp. 422–425, May 2003. doi: 10.1080/14620316.2003.11511642. Abstract
     Ashwagandha (Withania somnifera L. Dunal; Solanaceae) is being cultivated around the world mainly for its root which has rejuvenative properties. Field experiments during 1999-2001 under semi-arid tropical conditions of Hyderabad, India studied the effect of tillage depths (15 and 30 cm) and plant population densities (20, 40, 60 and 80 plants m-2) on root morphology, yield and quality and cultivation economics of ashwagandha. Preparatory tillage to 30 cm depth in combination with a density of 60 plants m-2 gave the highest root yield of 1.2 t ha-1 which was 50% higher than that following 15 cm tillage depth and the same density. This is attributed to the favourable effect of the former treatment leading to 33, 37 and 21% increases in plant height, number of branches per plant and shoot biomass yield, respectively. Further, this treatment produced most (68%) best quality root pieces. Main root length, length of lateral roots and diameter of lateral roots were significantly increased, while the diameter of main root and the number of laterals decreased. Furthermore, increase in plant density from 20-80 plants m-2 increased production of best quality roots from 42% to 59% under shallow tillage and from 53% to 71% of root yield under deep tillage. The implications of change in root morphology are discussed in the light of production of different grades of roots, their market preferences and economics. Deep tillage was equally advantageous for seed production, but plant population density for maximum seed yield (211 kg ha-1) was lower 40 than at 60 plants m-2 for maximum root yield. Maximum gross and net returns and benefit-cost ratio occurred following the highest root yield.
278. M. A. Kravchenko, “Effect of fertilizers containing potassium and chlorine on the crop and quality of Nicotiana rustica.,” Tabachnaya Promyshlennost’ SSSR, vol. 10, no. 9, pp. 26–27, 1940. https://www.cabdirect.org/cabdirect/abstract/19431901204. Abstract
     KC1, when used in proper proportion with N and P fertilizers, does not affect flavour and burning properties.
279. B. Krock, S. Schmidt, C. Hertweck, and I. T. Baldwin, “Vegetation-Derived Abscisic Acid and Four Terpenes Enforce Dormancy in Seeds of the Post-Fire Annual, Nicotiana Attenuata,” Seed Science Research, vol. 12, no. 4, pp. 239–252, Dec. 2002. doi: 10.1079/SSR2002117. Abstract
     The native tobacco, Nicotiana attenuata, synchronizes its germination with the immediate post-fire environment with a combination of germination stimulants found in wood smoke and inhibitors from the unburned litter of the dominant vegetation. The inhibitors override the stimulants and prevent seeds from germinating maladaptively in unburned habitats adjacent to burns. To understand the physiological basis of this environmental control of germination, we tested several previously isolated signals, phytohormones and their respective biosynthesis inhibitors. The germination inhibitors methyl jasmonate (MeJA, a constituent of sagebrush litter), bornane-2,5-dione (BD, a constituent of juniper litter extract, JLE) and JLE did not alter abscisic acid (ABA) content of imbibed seeds. Treatment with the ABA biosynthesis inhibitor, fluridone, inhibited the dormancy-inducing effects of BD, JLE and MeJA, but surprisingly did not affect endogenous ABA levels in treated seeds. However, ABA leached from litter of the species, which dominate the plant community before fires, plays an important role in germination control. We conclude that N. attenuata seeds, which can lie dormant in the soil for 150 years between fires, time their germination with the post-fire environment by responding to smoke, ABA and four terpenes (BD, 1,8-cineole, β-thujaplicin and camphor) leaching from the litter of the dominant vegetation.
280. V. S. Kubsad, Y. B. Palled, and C. P. Mansur, “Effect of Spacing and Fertilizer Levels on Physiological Parameters in Relation to Productivity of Ashwagandha (Withania Somnifera).,” Indian Journal of Agricultural Sciences, vol. 79, no. 7, pp. 501–505, 2009. https://www.cabdirect.org/cabdirect/abstract/20093217565. Abstract
     A field experiment was conducted during rabi seasons of 2004 and 2005 at Agricultural Research Station, Annigeri, Karnataka, to study the effect of spacings and fertilizer levels on physiological parameters in relation to productivity of ashwagandha (Withania somnifera Dunal) in Vertisols. Sixteen treatments comprising 4 spacings (15 cm × 5 cm, 15 cm × 10 cm, 30 cm × 10 cm and 45 cm × 10 cm) and...
281. V. S. Kubsad, Y. B. Palled, C. P. Mansur, H. T. Channal, N. Basavaraj, and R. V. Koti, “Performance of Ashwagandha (Withania Somnifera Dunal) as Influenced by Dates of Sowing and Stages of Harvesting,” p. 5, 2004. Abstract
     A field experiment was conducted during late kharif seasons of 2004-05 and 2005-06 to find out an optimum date of sowing and stage of harvesting to maximize productivity and enhance the quality of ashwagandha in vertisols under rainfed conditions at Agricultural Research Station, Annigeri (Karnataka). There were twelve treatments consisting of four date of sowing ( Aug. 15th , Aug. 30th , Sep. 15th and Sep. 30th ) and three stage of harvesting (120, 150 and 180 DAS). Pooled results over two years indicated that crop sown on Sep 15th gave more dry root yield by 11.9, 9.1 and 6.3% and net returns by 14.5, 11.2 and 7.8% than Aug 15th , Aug 30th and Sep 30th sowings respectively. While the late harvested crop at 180 DAS gave significantly higher dry root yield by 28.2 and 18.8% over early harvested crop at 120 and 150 DAS respectively. This treatment also provided the highest net returns (Rs. 48,430/ha) compared to others. The total withanolide content and total withanolide yield were maximum with Sep 15th sowing (0.537% and 7.233 kg/ha respectively) and also with late harvested crop (0.560% and 8.181 kg/ha respectively) compared to other sowing dates and harvesting stages. Similar trend was observed during both the years.
282. A. A. Kulkarni, S. M. Kelkar, M. G. Watve, and K. V. Krishnamurthy, “Characterization and Control of Endophytic Bacterial Contaminants in in Vitro Cultures of Piper Spp., Taxus Baccata Subsp. Wallichiana, and Withania Somnifera,” Canadian Journal of Microbiology, vol. 53, no. 1, pp. 63–74, Jan. 2007. doi: 10.1139/w06-106. Abstract
     Bacterial contamination is a serious problem that causes severe loss of in vitro grown cultures of a number of plants. This problem becomes even more acute if the bacterial contamination is of endophytic origin. In such cases, identification and characterization of the contaminants is essential for achieving specific control of the contaminants through selective use of antibiotic agents, especially if the routinely used contamination control methods practiced elsewhere in tissue culture studies are ineffective. Such is the case with the bacterial contamination observed in the present study. The five endophytic bacteria associated with Piper nigrum and Piper colubrinum, four endophytic bacteria associated with Taxus baccata subsp. wallichiana, two endophytic bacteria associated with Withania somnifera, and two bacteria common to all these plant species were isolated and characterized based on morphological and biochemical tests. Their taxonomic positions based on similarity indices were determined. A control strategy against these bacteria has been developed based on bacteriostatic or bactericidal actions of 12 antibiotics at three different concentrations by solid and liquid antibiogramme assays.Key words: antibiotics, endophytic bacterial contamination, identification, Piper nigrum, Piper colubrinum, Taxus baccata subsp. wallichiana, Withania somnifera.
283. A. Kumar et al., “Biocontrol Potential of Pseudomonas Stutzeri Endophyte from Withania Somnifera (Ashwagandha) Seed Extract against Pathogenic Fusarium Oxysporum and Rhizoctonia Solani,” Archives of Phytopathology and Plant Protection, vol. 55, no. 1, pp. 1–18, Jan. 2022. doi: 10.1080/03235408.2021.1983384. Abstract
     Agricultural crops are susceptible to abiotic and biotic factors. Among biotic factors, plant pathogens attacks are prominent and significantly reduce the crop productivity and production. Long term application of synthetic pesticides in agriculture not only causes resistance development in pathogens but also deteriorates the soil quality and pollutes the ecosystem. In the current study, an investigation has been made with an objective to isolate an endophytic bacterium from Withania somnifera seed and evaluate its potential biocontrol activity against soil-borne pathogens like Fusarium oxysporum var. ciceri and Rhizoctonia solani. The molecular approach of bacterium identification using 16S rRNA sequencing methods includes kit based DNA extraction and PCR amplification and the isolated endophytic bacterium was identified as Pseudomonas stutzeri. In dual culture assay, the endophytic bacterium P. stutzeri PSIISS-1 effectively suppressed growth of pathogenic fungi F. oxysporum var. ciceri (61.0%) and R. solani (53.3%), respectively. In vivo studies were conducted with initial microbial inoculum size of 108cell/ml and the studies revealed that the bacterium P. stutzeri PSIISS-1 co-inoculated with R. solani had significantly inhibited the sheath blight disease with the efficacy of 76% in susceptible paddy MTU-7029. Simultaneously, co-inoculation of P. stutzeri PSIISS-1 along with the pathogen F. oxysporum var. ciceri in chickpea JG-14 variety resulted in reduction of wilt symptom up to 80.45%. Bacterial inhibitions against the pathogens were corroborated with chitinase, protease and lipase enzymatic activities. Based on the results obtained from the study, P. stutzeri showed biocontrol activity against plant pathogens which can be used as potential bio-pesticides.
284. M. Kumar et al., “Delineating G × E Interactions by AMMI Method for Root Attributes in Ashwagandha [Withania Somnifera (L.) Dunal],” INDIAN JOURNAL OF GENETICS AND PLANT BREEDING, vol. 80, no. 04, pp. 441–449, Dec. 2020. doi: 10.31742/IJGPB.80.4.10. Abstract
     In the present study, additive main effects and multiplicative interactions (AMMI) biplot analyses was used to dissect genotype x environment interaction (GEI) and to identify location specific and widely adapted genotypes for root branches, diameter and length in ashwagandha [Withania somnifera (L.) Dunal]. Trials were conducted in randomized complete block design (RCBD) with two replications over three consecutive years at three different locations. ANOVA analysis revealed environment, G×E interaction and genotype effects to contribute significantly (p less than 0.001) towards total sum of squares for root branches (61.00%, 22.18% and 14.00%); root diameter (51.06%, 24.26% and 15.34%) and root length (65.67%, 20.82% and 11.39%). Further, the GEI for these traits was mostly explained by the first, second and third principal component axis (IPCA1, IPCA2 and IPCA3). AMMI1 and AMMI2 biplot analyses showed differential stability of genotypes for root branches, diameter and length with few exceptions. Environmental contribution towards the genotypic performance from AMMI1 and AMMI2 analysis for root traits except environment Bhi16 contribution for root diameter and root length. AMMI1 biplots and simultaneous selection index (SSI) statistics identified SKA-11 as the most desirable genotype for root branches and length while SKA-26 and SKA-27 for root diameter. The ashwagandha genotypes identified for root attributes could be advocated either for varietal recommendation or in varietal development program.
285. \relax M. I. T. H. L. E. S. H. KUMAR et al., “Elucidation of Genotype–Environment Interactions and Genetic Stability Parameters for Yield, Quality and Agromorphological Traits in Ashwagandha (Withania Somnifera (L.) Dunal),” Journal of Genetics, vol. 99, no. 1, p. 59, Jul. 2020. doi: 10.1007/s12041-020-01207-9. Abstract
     The present study was undertaken to delineate genotype–environment interactions and stability status of 16 genotypes of ashwagandha (Withania somnifera (L.) Dunal) in context to the 12 characters, namely plant height, number of primary branches, number of secondary branches, days to flowering, days to maturity, number of berries, number of seeds/berry, root length, root diameter, root branches, dry root yield and total alkaloid content (%). Experiment was carried out in a randomized complete block design with three replications over three different locations (S. K. Nagar, Jagudan and Bhiloda) in north Gujarat for three years (2016–17, 2017–18 and 2018–19). Pooled analysis of variance revealed that the mean squares due to genotypes and genotype\,× environment interaction along with linear and nonlinear components were highly significant (P<0.01) for most of the traits under study. Stability parameters for component traits through Eberhart and Russell model showed that genotypes that can be used directly in breeding programme are SKA-4 for early flowering, SKA-21 for early maturity and SKA-1, SKA-4, SKA-6 and SKA-17 for shorter plant height. Further, SKA-21 could be used for improving number of primary branches per plant, SKA-11 and SKA-17 for number of secondary branches per plant, SKA-19 for number of berries per plant, SKA-6, SKA-21, SKA-27 and AWS-1 for root branches and SKA-17 for root length as these genotypes were found to be most stable across the environments for mentioned traits. The result revealed that some reliable predictions about genotype\,× environment interaction and its unpredictable components were involved significantly in determining the stability of genotypes. Hence, the present investigation can be exploited for the identification of more productive genotypes in specific environments, leading to significant increase in root productivity of ashwagandha.
286. A. Kumar et al., “Exegesis of Cultivars-Multi-Year/Environment Interactions for Agro Morphological Traits in Ashwagandha (Withania Somnifera (L.) Dunal),” South African Journal of Botany, vol. 151, pp. 523–531, Dec. 2022. doi: 10.1016/j.sajb.2022.10.039. Abstract
     A randomized block design was used to examine the important medicinal plant ashwagandha in three replications throughout the late Kharif seasons of 2018, 2019, and 2020. The cultivars of Ashwagandha (Withania somnifera Dunal), CIMAP Chetak, CIMAP Pratap, NMITLI-101, NMITLI-118, and Poshita were used to evaluate the quality of dry root yield for a better quality of five variations over the years in south India and variety recommendations. The years I and III were the two years with the most discrimination, whereas year II had the least discrimination. A lower AEA angle in a test year is more typical of subsequent test years. The first and second years are the most representative, whereas the third and fourth are the least representative. The year I was discriminating and representative during the test year, making them suitable candidates for selecting adapted varieties. The separation of stable and unstable types was made easier in environments that were discriminating or non-representative of test years. There was much diversity for stability in the various traits, as reflected by the character means. A line that passes through the biplot origin and measures performance over a year determines the rank. Four varieties, CIMAP-Pratap, Poshtita, CIMAP-Chetak, and NMITLI-118, were highly stable and good rankers for the dry root yield, root starch (%), starch and fiber ratio, and total alkaloid, among other economic traits (%). These varieties were highly stable for dry root yield and root starch (%), starch and fiber ratio, and total alkaloid, based on the mean performance of major economic features and stability measures (%). As a result, these types are recommended for further cultivation in southern India.
287. R. R. Kumar, A. P. L. Reddy, C. J. Subbaiah, N. A. Kumar, N. H. N. Prasad, and B. Bhukya, “Genetic Association among Root Morphology, Root Quality and Root Yield in Ashwagandha (Withania Somnifera),” Genetika, vol. 43, no. 3, pp. 617–624, 2011. doi: 10.2298/GENSR1103617K. Abstract
     Ashwagandha (Withania somnifera) is a dryland medicinal crop and roots are used as valuable drug in traditional systems of medicine. Morphological variants (morphotypes) and the parental populations were evaluated for root - morphometric, quality and yield traits to study genetic association among them. Root morphometric traits (root length, root diameter, number of secondary roots/ plant) and crude fiber content exhibited strong association among them and showed significant positive genotypic correlation with yield. Starch-fiber ratio (SFR), determinant of brittle root texture showed strong negative association with root yield. The total alkaloid content had positive genotypic correlation with root yield. So genetic upgradation should aim at optimum balance between two divergent groups of traits i.e. root yield traits (root morphometric traits and crude fiber content) and root textural quality traits (starch content and SFR) to develop superior genotypes with better yield and quality.
288. S. Kumar, N. Kaushik, and P. Proksch, “Identification of Antifungal Principle in the Solvent Extract of an Endophytic Fungus Chaetomium Globosum from Withania Somnifera,” SpringerPlus, vol. 2, no. 1, p. 37, Feb. 2013. doi: 10.1186/2193-1801-2-37. Abstract
     Extracts of Chaetomium globosum EF18, isolated as endophytic fungus from Withania somnifera, were found effective against Sclerotinia sclerotiorum. Ethyl acetate and methanol extracts were more effective than hexane extract showing >80% growth inhibition. Bioactive compound (antibiotic Sch 210971, m/z 445 and λmax 290) having antifungal activity against S. sclerotiorum has been isolated in pure form from the ethyl acetate extract following bioassay guided fractionation. Apart from this compound other fractions of polar to medium polarity were also found effective. Fraction no. VIII from VLC (Vacuum liquid chromatography) column of ethyl acetate extract was most active having IC50 value 35.4 μg/ml.
289. M. Kumar, K. Kumar, N. Kumar, and A. patel, “Impact of Different Manures on Ashwagandha (Withania Somnifera Dunal.) Production under Rainfed Condition,” International Journal of Current Microbiology and Applied Sciences, vol. 6, no. 10, pp. 50439–5059, Oct. 2017. doi: 10.20546/ijcmas.2017.610.479.
290. A. Kumar, M. Ali, and S. R. Mir, “A New Withanolide from the Roots of Withania Somnifera,” IJC-B Vol.43B(09) [September 2004], Sep. 2004. http://nopr.niscpr.res.in/handle/123456789/21335. Abstract
     Phytochemical studies on the roots of Withania somnifera (Solanaceae) resulted in the isolation of a new withanolide glycoside, glucosomniferanolide, characterized as (20R, 22S)- 1-oxo-witha-2, 5, 24-trienolide-20-β-ol-20-D-glucopyranoside by spectral analyses and chemical reactions.
291. A. Kumar, A. C. Jnanesha, V. Kumar M, and R. K. Lal, “Quantification of Genotype × Environment Interaction of Agronomic Traits in Ashwagandha (Withania Somnifera (L.) Dunal),” Acta Ecologica Sinica, Jul. 2022. doi: 10.1016/j.chnaes.2022.07.002. Abstract
     Ashwagandha is a popular industrially medicinal plant used in Ayurveda and other traditional medical systems. We looked at the stability of the sixteen economic traits in the five ashwagandha genotypes in four locations over two years in south India. The pooled ANOVA showed significant differences among different lines of ashwagandha. The five genotypes (G), genotypes × locations interaction (G × L), and treatments were highly significant for all sixteen traits. The character’s associations directly impact the genotype’s ability to sustain stability. The traits X10-VS-X13 and X3-VS-X4 were positively and significantly related across the locations over the three years, according to correlations among the selected traits, indicating that choosing one trait had a beneficial impact on its associated attributes. Except for location I year two, the traits, specifically X11-VS-X13, were highly significant across all locations. The characteristics, specifically X12-VS-X13, were also highly significant, except for location I year I. The genotypes nearest the ideal variety are more desirable than the other. The genotypes W.S. 3 and W.S. 5 for the traits X11 and X16, W.S. 2, and W.S. 5 for the trait X15 were more desirable for selection. In a large amount, both predictable and unanticipated components contributed to the stability. The PCA distribution also showed a significant amount of morphologically based genetic variability. Different genotypes of ashwagandha showed a substantial variation in X15 = Withaferin-A (%) and X16 = Withanolide-A (%). The percentages of X15 = Withaferin-A (W.S.2 > W.S.5 > W.S. > 4 W.S.3 > W.S.1) and X16 = Withanolide-A (W.S.3 > W.S.5 > W.S.4 > W.S.2 > W.S.1) were shown to be 0.07 to 1.24% and 0.088 to 1.88%, respectively. Based on the mean performance of the most economically important traits and stability parameters, the two superior genotypes, W.S. 3 and W.S. 5 were highly stable for dry root yield with high-quality alkaloids. In south India, these genotypes are recommended for further cultivation.
292. R. Kumar, A. J. Das, and A. A. Juwarkar, “Reclamation of Petrol Oil Contaminated Soil by Rhamnolipids Producing PGPR Strains for Growing Withania Somnifera a Medicinal Shrub,” World Journal of Microbiology and Biotechnology, vol. 31, no. 2, pp. 307–313, Feb. 2015. doi: 10.1007/s11274-014-1782-1. Abstract
     Soil contaminated by hydrocarbons, cannot be used for agricultural intents due to their toxic effect to the plants. Surfactants producing by plant growth promotory rhizobacteria (PGPR) can effectively rig the problem of petroleum hydrocarbon contamination and growth promotion on such contaminated soils. In the present study three Pseudomonas strains isolated from contaminated soil identified by 16S rRNA analysis were ascertained for PGPR as well as biosurfactants property. Biosurfactants produced by the strains were further characterized and essayed for rhamnolipids. Inoculation of the strains in petrol hydrocarbon contaminated soil and its interaction with Withania somnifera in presence of petrol oil hydrocarbons depict that the strains helped in growth promotion of Withania somnifera in petrol oil contaminated soil while rhamnolipids helped in lowering the toxicity of petrol oil. The study was found to be beneficial as the growth and antioxidant activity of Withania sominfera was enhanced. Hence the present study signifies that rhamnolipids producing PGPR strains could be a better measure for reclamation of petrol contaminated sites for growing medicinal plants.
293. A. Kumar et al., “Utility of a Multidisciplinary Approach for Genome Diagnostics of Cultivated and Wild Germplasm Resources of Medicinal Withania Somnifera, and the Status of New Species, W. Ashwagandha, in the Cultivated Taxon,” Plant Systematics and Evolution, vol. 291, no. 3, pp. 141–151, Feb. 2011. doi: 10.1007/s00606-010-0372-4. Abstract
     Realizing the inconsistencies that exist in the extent and nature of differentiation in the Withania somnifera genetic resources in India, the 21 cultivated and wild accessions, and the two hybrids (cultivated × wild accessions and vice versa) were investigated for morphological, cytogenetical, chemical profiling, and crossability features. Their nuclear and chloroplast genomes were also assayed at the nucleotide sequence level, and by use of DNA markers. Chloroplast DNA diversity and somatic chromosome number (2n = 48) were not helpful in identifying the differences. Other approaches, on the other hand, especially restriction endonuclease digests, types and sequence length composition of ITS 1 and ITS 2 of nuclear ribosomal DNA, AFLP fingerprinting, and crossability barriers unambiguously provided startling discrete differences between the cultivated and wild accessions, indicating a clear division of W. somnifera into two distinct lineages. These data, therefore, are indicative of the fact that because of the unique characteristics of its nuclear genome, and strong crossability barriers vis-à-vis wild accessions of W. somnifera, the cultivated accessions should be relegated to the rank of the separate species, W. ashwagandha.
294. \relax V. I. V. E. K. KUMAR, \relax A. M. A. R. SOLANKI, and \relax S. H. I. V. E. S. H. SHARMA, “Yield and Economics of Withania Somnifera Influenced by Dual Inoculation of Azotobacter Chroococcum and Pseudomonas Putida,” Turkish Journal of Biology, vol. 33, no. 3, pp. 219–223, Jan. 2009. doi: 10.3906/biy-0711-7. Abstract
     The yield parameters and cost economics of Withania somnifera were studied using dual inoculation of Azotobacter chroococcum and Pseudomonas putida. The experiment was conducted in a randomized block design with 3 replications for 2 consecutive years, along with 2 levels of organic manure, 10 t ha−1 and 20 t ha−1. Inoculation with the inoculants generated encouraging results; root yield (1185.6 kg ha−1), seed yield (208.13 kg ha−1), number of offshoots per plant (6.07), and plant height (108.4 cm) were maximum with treatment T6 (organic manure [OM] 20 t ha−1 + both bacteria), followed by T9 (OM 10 t ha−1 + both bacteria), T4 (OM 20 t ha−1 + A. chroococcum), and T5 (OM 20 t ha−1 + P. putida), as compared to T1 (control), T2 (OM 10 t ha−1), and T3 (OM 20 t ha−1). The survival count of inoculated bacteria was highest 70 days after inoculation and declined thereafter. Economic analysis revealed that the net profit from treatment T6 was the highest with dual inoculation of bacteria and OM 20 t ha−1, followed by OM 10 t ha_1 and dual inoculation of bacteria.
295. R. K. Kushwaha et al., “Compatibility of Inherent Fungal Endophytes of Withania Somnifera with Trichoderma Viride and Its Impact on Plant Growth and Withanolide Content,” Journal of Plant Growth Regulation, vol. 38, no. 4, pp. 1228–1242, Dec. 2019. doi: 10.1007/s00344-019-09928-7. Abstract
     Withania somnifera (Ashwagandha), also known as Indian ginseng, is an important ancient medicinal plant, used in the Indian traditional systems of medicine. In view of increasing demand for roots of Ashwagandha, the present study was undertaken to investigate the compatibility of inherent fungal endophytes along with the biocontrol agent, Trichoderma viride, for enhancing W. somnifera plant growth and root secondary metabolites (withaferin A). It has frequently been emphasized by the World Health Organization the use of healthy roots of Ashwagandha for therapeutic applications. To maintain quality of W. somnifera roots, an option could be eco-friendly management of root-knot diseases and co-inoculation of native endophytes along with T. viride. The in vitro antagonistic activity of T. viride (TV) against the W. somnifera pathogens, Alternaria alternata and Sclerotium rolfsii, showed 64.3% and 69.5% growth inhibition, respectively. Here, we investigated the compatibility of TV along with the native endophytic fungi Aspergillus terreus strain 2aWF (2aWF), Penicillium oxalicum strain 5aWF (5aWF), and Sarocladium kiliense strain 10aWF (10aWF) for the cultivation of W. somnifera. The co-inoculation of TV and native endophytic fungi resulted in increased shoot, root weight, and plant height to 65–150%, 35–74.5%, and 15–35%, respectively, compared to untreated plants. Withanolide A content in leaves of TV-treated plants increased significantly by 260%, whereas in co-inoculation treatments, it was enhanced up to 109–242%. However, no considerable change was noticed with withaferin A content in leaves, except the 2aWF\,+ TV treatment significantly increased by 27%. In contrast, withanolide A content in roots was not affected by TV alone but co-inoculation with endophyte treatments significantly increased its content (19–73%). TV alone had increased chlorophyll a by 23%; however, in combination treatments, it increased up to 115–164% compared to control. Besides secondary metabolites in roots and leaves, co-inoculation of TV and native endophytes modulated the expression of the withanolide biosynthetic pathway genes HMGR, DXR, FPPS, SQS, SQE, CAS, SMT1, STE1, and CYP710A1 compared to control treatments. Apart from withanolide biosynthetic pathway genes, co-inoculation of TV also ameliorated the host-resistant-related gene NPR1 which was upregulated by ninefold in the TV treatment and 3- to 7-fold in the combination treatment. Overall, our results show that co-inoculation of TV along with inherent endophytes of W. somnifera enhanced plant growth and withanolides accumulation.
296. G. A. Kutlymuratova, “Ecological Features and Agrotechnology of Growing Atropa Belladonna l. and Echinops Ritro L in the Conditions of Karakalpakstan,” Journal of Pharmaceutical Negative Results, pp. 2400–2404, Nov. 2022. doi: 10.47750/pnr.2022.13.S08.297. Abstract
     The article deals with the study of ecological features and agrotechnology of growing Atropa belladonna L. and Echinops ritro L in the conditions of the Republic of Karakalpakstan. The adaptive adaptability is the highest in Atropa belladonna L., the plant has a high combined resistance to local climatic conditions, massively blooms and bears fruit, actively self-propagates vegetatively.
297. M. Kyo and H. Harada, “Phosphorylation of Proteins Associated with Embryogenic Dedifferentiation of Immature Pollen Grains of Nicotiana Rustica,” Journal of Plant Physiology, vol. 136, no. 6, pp. 716–722, Oct. 1990. doi: 10.1016/S0176-1617(11)81351-9. Abstract
     Pollen embryogenesis in Nicotiana rustica can be induced by starvation of immature pollen grains at a specific developmental stage.Using a highly homogeneous population of isolated pollen grains at the mid-bicellular stage, we observed embryogenic cell division at a high frequency, which allowed us to carry out a biochemical investigation of events that occur during the initial period of pollen embryogenesis.Two-dimensional gel electrophoresis and autoradiography revealed a pattern of proteins phosphorylated in vivo, which consisted of six spots and was specific to embryogenic pollen grains.This characteristic pattern of phosphorylation was observed neither with pollen grains engaged in normal development (maturation) nor with non-embryogenic pollen derived from pollen grains younger or older than those at the mid-bicellular stage.The observed pattern of phosphorylation may be closely related to the initiation of pollen embryogenesis.
298. B. Ladešić, M. Pokorny, and D. Keglević, “Evidence for D-Configuration of N-Malonylmethionine, the Metabolite of d-Methionine in Nicotiana Rustica,” Phytochemistry, vol. 9, no. 10, pp. 2105–2107, Oct. 1970. doi: 10.1016/S0031-9422(00)85374-1. Abstract
     Evidence for the d-configuration of N-malonylmethionine-methyl-14C isolated from tobacco plant fed with d-methionine-methyl-14C was obtained by acid hydrolysis of the conjugate and subsequent deamination of methionine with d-amino acid oxidase. Parallel experiments with synthetically prepared N-malonyl-d-methionine were carried out.
299. R. K. Lal, “Quantification of Adaptability and Stability among Genotypes/Cultivars for Root Yield in Ashwagandha (Withania Somnifera L.),” Industrial Crops and Products, vol. 77, pp. 648–657, Dec. 2015. doi: 10.1016/j.indcrop.2015.09.035. Abstract
     Ashwagandha (Withania somnifera L.), is one of the very important medicinal plant in Ayurveda since ancient times. It is a native of Mediterranean region grows wildly in arid and semi arid parts of India. Under the germplasm enhancement programme, thirty genetic stocks/cultivars of ashwagandha originated from selective divergence belongs to eight states of India (U.P., M.P., Rajasthan, Maharashtra, Kerala, J and K., A.P. and W.B.) were evaluated in the four consecutive years for the estimation of stability/adaptability. Using the AMMI model, ashwagandha genetic stocks namely, W 20, W 1 (cv. Pratap), W 2, W 3 (cv. Chetak), W 4 and W 6 (cv.Poshita) were expressed the high adaptability and stability for root yield over years. Therefore, above six selected genetic stocks/varieties recommended for commercial cultivation in India on large scale.
300. W. Lammerts, “Interspècific Hybridization in Nicotiana. IX. Further Studies of the Cytology of the Backcross Progenies of the Paniculata-Rustica Hybrid,” Genetics, vol. 14, no. 3, pp. 286–304, May 1929. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1201036/.
301. C. Laszlo et al., “Fractionation and Extraction Optimization of Potentially Valuable Compounds and Their Profiling in Six Varieties of Two Nicotiana Species,” Molecules, vol. 27, no. 22, p. 8105, Jan. 2022. doi: 10.3390/molecules27228105. Abstract
     There is an increasingly urgent call to shift industrial processes from fossil fuel feedstock to sustainable bio-based resources. This change becomes of high importance considering new budget requirements for a carbon-neutral economy. Such a transformation can be driven by traditionally used plants that are able to produce large amounts of valuable biologically relevant secondary metabolites. Tobacco plants can play a leading role in providing value-added products in remote areas of the world. In this study, we propose a non-exhaustive list of compounds with potential economic interest that can be sourced from the tobacco plant. In order to optimize extraction methodologies, we first analyzed their physico-chemical properties using rapid solubility tests and high-resolution microfractionation techniques. Next, to identify an optimal extraction for a selected list of compounds, we compared 13 different extraction method–solvent combinations. We proceeded with profiling some of these compounds in a total of six varieties from Nicotiana tabacum and Nicotiana rustica species, identifying the optimal variety for each. The estimated expected yields for each of these compounds demonstrate that tobacco plants can be a superior source of valuable compounds with diverse applications beyond nicotine. Among the most interesting results, we found high variability of anatabine content between species and varieties, ranging from 287 to 1699 µg/g. In addition, we found that CGA (1305 µg/g) and rutin (7910 µg/g) content are orders of magnitude lower in the Burley variety as compared to all others.
302. D. Lavie, E. Glotter, and Y. Shvo, “1371. Constituents of Withania Somnifera Dun. Part IV. The Structure of Withaferin A,” Journal of the Chemical Society (Resumed), no. INVALID_SCITE_VALUE, pp. 7517–7531, Jan. 1965. doi: 10.1039/JR9650007517. Abstract
     No abstract available
303. D. Lavie, S. Greenfield, and E. Glotter, “Constituents of Withania Somnifera Dun. Part VI. The Stereochemistry of Withaferin A,” Journal of the Chemical Society C: Organic, no. INVALID_SCITE_VALUE, pp. 1753–1756, Jan. 1966. doi: 10.1039/J39660001753. Abstract
     Withaferin A, the steroidal lactone isolated from the leaves of Withania somnifera Dun. (Solanaceae), has been degraded to bisnor-(5α)-cholanic acid. The β-orientation of the 5,6-epoxide has been demonstrated by optical rotatory dispersion and n.m.r. data. Consequently, withaferin A is a 5β-steroid with the usual stereochemistry at all asymmetric centres, possessing a six-membered-ring lactone in the side-chain.
304. D. Lavie, I. Kirson, and E. Glotter, “Constituents of Withania Somnifera Dun. Part X The Structure of Withanolide D,” Israel Journal of Chemistry, vol. 6, no. 5, pp. 671–678, 1968. doi: 10.1002/ijch.196800085. Abstract
     Withanolide D, the major component of the leaves of Withania somnifera chemotype II, is a steroidal lactone of the withanolide type, isomeric with withaferin A. It has been identified as 5, 6β-epoxy-4β, 20α-dihydroxy-1-oxo-(5β)witha-2, 24-dienolide. The structure has been elucidated by NMR measurements and by chemical degradation to 4β-acetoxy-5,6β-epoxy (5β) pregnan-1, 20-dione.
305. D. Lavie, E. Glotter, and Y. Shvo, “Constituents of Withania Somnifera Dun. III. The Side Chain of Withaferin A*,1,” ACS Publications. American Chemical Society, May-2002. doi: 10.1021/jo01017a015.
306. J. Layton, “Improvements to Solanum Tuberosum Cv. ‘Desireé’ Cell Suspension Culture and Nicotiana Tabacum Cv. ‘Petit Havana’ Shoot Induction.,” Masters Theses, Dec. 2020. https://trace.tennessee.edu/utk_gradthes/5848.
307. S.-H. Lee, R. K. Tewari, E.-J. Hahn, and K.-Y. Paek, “Photon Flux Density and Light Quality Induce Changes in Growth, Stomatal Development, Photosynthesis and Transpiration of Withania Somnifera (L.) Dunal. Plantlets,” Plant Cell, Tissue and Organ Culture, vol. 90, no. 2, pp. 141–151, Aug. 2007. doi: 10.1007/s11240-006-9191-2. Abstract
     The aim of the study was to establish whether the quantity and the quality of light affect growth and development of Withania somnifera plantlets. We have studied growth and histo-physiological parameters [stomatal characteristics, chloroplastic pigments concentrations, photosynthesis, and transpiration (E)] of W. somnifera plantlets regenerated under various light intensities, or monochromatic light or under a mixture of two colors of light in tissue culture conditions. Plantlets grown under a photon flux density (PFD) of 30 μmol m-2 s-1 showed greater growth and development than those raised under other PFDs. Chlorophylls and carotenoids, numbers of stomata, rate of photosynthesis (PN) and transpiration (E), stomatal conductance (gs), and water use efficiency (WUE) increased with increasing the PFD up to 60 μmol m-2 s-1. Light quality also affected plantlets growth and physiology. Highest growth was observed under fluorescent and in a mixture of blue and red light. Very few stomata were developed in any of the monochromatic light but under fluorescent or under a mixture of two colors stomatal numbers increased. Similarly, gs, E, PN, and WUE were also higher under fluorescent light and under a mixture of red and blue light. Regressional analysis showed a linear relationship between PN (r2 = 70) and gs and between E (r2 = 0.95) and gs. In conclusion, both the quality and the quantity of light affect growth of plantlets, development of stomata and physiological responses differently depending on the intensity and the wavelength of light.
308. W. Lein et al., “Large-Scale Phenotyping of Transgenic Tobacco Plants (Nicotiana Tabacum) to Identify Essential Leaf Functions,” Plant Biotechnology Journal, vol. 6, no. 3, pp. 246–263, 2008. doi: 10.1111/j.1467-7652.2007.00313.x. Abstract
     Two of the major challenges in functional genomics are to identify genes that play a key role in biological processes, and to elucidate the biological role of the large numbers of genes whose function is poorly characterized or still completely unknown. In this study, a combination of large-scale expressed sequence tag sequencing, high-throughput gene silencing and visual phenotyping was used to identify genes in which partial inhibition of expression leads to marked phenotypic changes, mostly on leaves. Three normalized tobacco (Nicotiana tabacum) cDNA libraries were prepared directly in a binary vector using different tissues of tobacco as an RNA source, randomly sequenced and clustered. The Agrobacterium–tobacco leaf disc transformation system was used to generate sets of antisense or co-suppression transgenic tobacco plants for over 20 000 randomly chosen clones, each representing an independent cluster. After transfer to the glasshouse, transgenic plants were scored visually after 10–14 days for changes in growth, leaf form and chlorosis or necrosis. Putative hits were validated by repeating the transformation. This procedure is more stringent than the analysis of knockout mutants, because it requires that even a partial decrease in expression generates a phenotype. This procedure identified 88 validated gene/phenotype relations. These included several previously characterized gene/phenotype relationships, demonstrating the validity of the approach. For about one-third, a function could be inferred, but a loss-of-function phenotype had not been described previously. Strikingly, almost one-half of the validated genes were poorly annotated, or had no known function. For 77 of these tobacco sequences, a single or small number of potential orthologues were identified in Arabidopsis. The genes for which orthologues were identified in Arabidopsis included about one-half of the genes whose function was completely unknown. Comparison with published gene/phenotype relations for Arabidopsis knockout mutants revealed surprisingly little overlap with the present study. Our results indicate that partial gene silencing identifies novel gene/phenotype relationships, which are distinct from those uncovered by knockout screens. They also show that it is possible to perform these analyses in a crop species in which full genome sequence information is lacking, and subsequently to transfer the information to a reference species in which functional studies can be performed more effectively.
309. B. Lercari and F. Sponga, “Effect of Chemical Treatments on the Photocontrol of Germination in «Atropa Belladonna» L. Seeds,” Rivista di ortoflorofrutticoltura italiana, vol. 69, no. 2, pp. 115–122, 1985. https://www.jstor.org/stable/42878729. Abstract
     The effects of many chemicals (fusicoccin, GA₃, GA4/7, abscissic acid (ABA), cycloheximide and actinomycin D) on the light-mediated germination of the seeds of A. belladonna were studied. Fusicoccin cannot induce seed germination in the dark, but it enhances the percentage germination either in the dark or under continuous red light. It appears that the two factors red light and GA act independently. ABA and cycloheximide inhibit completely the light mediated germination, while the effect of actinomycin D is very low. It is suggested that phytochrome-mediated seed germination in A. belladonna requires cytoplasmic protein-synthesis, while there are no indications for a requirement in RNA synthesis. Our results exclude the likelihood that phytochrome controls germinations of Atropa belladonna seeds through the activation of a H⁺/K⁺ exchange mechanism, fusicoccin sensitive, at the membrane level. Si è studiato l’effetto di vari composti chimici (fusicoccina, GA₃, GA4/7, acido abscissico (ABA), cicloesimide e actinomicina D) sulla germinazione dei semi di A. belladonna indotta dalla luce. La fusicoccina non è in grado di indurre la germinazione al buio, ma la aumenta alla luce. GA₃ e GA4/7 sono in grado di stimolare la germinazione sia alla luce che al buio. I due fattori luce e gibberelline sembrano agire indipendentemente l’uno dall’altro nel controllo della germinazione di A. belladonna. L’acido abscissico e la cicloesimide inibiscono completamente la germinazione indotta dalla luce, mentre l’effetto della actinomicina D è scarso o nullo. I risultati suggeriscono che in A. belladonna la germinazione mediata dal fitocromo richiede sintesi proteica citoplasmatica e sembra non richiedere l’attivazione a livello di membrana di un meccanismo di scambio H⁺/K⁺ fusicoccina sensibile.
310. R. S. Lewis and J. S. Nicholson, “Aspects of the Evolution of Nicotiana Tabacum L. and the Status of the United States Nicotiana Germplasm Collection,” Genetic Resources and Crop Evolution, vol. 54, no. 4, pp. 727–740, Jun. 2007. doi: 10.1007/s10722-006-0024-2. Abstract
     The genus Nicotiana is a member of the nightshade (Solanaceae) family, and is comprised of 70 currently recognized, naturally occurring species. Genetic variability within N. tabacum L., the species of primary economic importance, was likely affected by several genetic bottlenecks. Nicotiana tabacum is a classic amphidiploid that arose after chance interspecific hybridization between N. sylvestris Spegazinni et Comes and a member of section Tomentosae, likely N. tomentosiformis Goodspeed, N. otophora Grisebach, or an introgressive hybrid between the two. Only a fraction of the genetic variability that existed in the diploid progenitor gene pools probably entered into N. tabacum. Genetic drift, coupled with natural and human selection, subsequently resulted in the formation of narrow genetic pools corresponding to modern commercial market classes. Genetic variability in Nicotiana has gained increased attention in recent years because of investment in Nicotiana genomics research, interest in development of tobacco products with reduced harm characteristics, and concentration on using Nicotiana species for plant-based production of commercially useful proteins. A storehouse of genetic diversity for N. tabacum is available in approximately 1,900 accessions maintained by the United States Nicotiana Germplasm Collection. Seeds of 224 accessions representing 59 wild Nicotiana species are also maintained. The collection is currently maintained by North Carolina State University and is part of the United States National Plant Germplasm System (NPGS). The collection’s curator satisfies hundreds of seed requests made annually by scientists using Nicotiana germplasm for basic biological investigations and by researchers in the area of applied tobacco science.
311. R. S. Lewis, “Nicotiana Tabacum L.: Tobacco,” in Medicinal, Aromatic and Stimulant Plants, J. Novak and W.-D. Blüthner, Eds. Cham: Springer International Publishing, 2020, pp. 345–375. doi: 10.1007/978-3-030-38792-1_9. Abstract
     Tobacco (Nicotiana tabacum L.) is cultivated primarily for use in the manufacture of nicotine-containing products such as cigarettes, cigars, and smokeless tobacco products. Plant breeding methodologies are used to develop new tobacco cultivars with important trait combinations that contribute to the economy of crop production and to preferred characteristics for product manufacturing. Desired traits for new cultivars include high cured leaf yields, disease resistance, satisfactory physical cured leaf quality, appropriate cured leaf chemistry, and acceptable sensory attributes. Types of variation that might be used in tobacco breeding programs include that native to N. tabacum or its close relatives. De novo variation generated via mutation breeding, gene editing, or genetic engineering might also be considered. Methodologies used to combine desired traits into new commercial inbred lines and hybrids may vary according to the breeding program.
312. Y. Li et al., “Cold Stress in the Harvest Period: Effects on Tobacco Leaf Quality and Curing Characteristics,” BMC Plant Biology, vol. 21, no. 1, p. 131, Mar. 2021. doi: 10.1186/s12870-021-02895-w. Abstract
     Weather change in high-altitude areas subjects mature tobacco (Nicotiana tabacum L.) to cold stress, which damages tobacco leaf yield and quality. A brupt diurnal temperature differences (the daily temperature dropping more than 20\,°C) along with rainfall in tobacco-growing areas at an altitude above 2450 m, caused cold stress to field-grown tobacco.
313. Z.-G. Li, M. Gong, H. Xie, L. Yang, and J. Li, “Hydrogen Sulfide Donor Sodium Hydrosulfide-Induced Heat Tolerance in Tobacco (Nicotiana Tabacum L) Suspension Cultured Cells and Involvement of Ca2+ and Calmodulin,” Plant Science, vol. 185–186, pp. 185–189, Apr. 2012. doi: 10.1016/j.plantsci.2011.10.006. Abstract
     Hydrogen sulfide (H2S) is considered as a new emerging cell signal in higher plants. Hydrogen sulfide donor, sodium hydrosulfide, pretreatment significantly increased survival percentage of tobacco suspension cultured cells under heat stress and regrowth ability after heat stress, and alleviated decrease in vitality of cells, increase in electrolyte leakage and accumulation of malondialdehyde (MDA). In addition, sodium hydrosulfide-induced heat tolerance was markedly strengthened by application of exogenous Ca2+ and its ionophore A23187, respectively, while this heat tolerance was weakened by addition of Ca2+ chelator ethylene glycol-bis(b-aminoethylether)-N,N,N′,N′-tetraacetic acid (EGTA), plasma membrane channel blocker La3+, as well as calmodulin (CaM) antagonists chlorpromazine (CPZ) and trifluoperazine (TFP), respectively, but intracellular channel blocker ruthenium red (RR) did not. These results suggested that sodium hydrosulfide pretreatment could improve heat tolerance in tobacco suspension cultured cells and the acquisition of this heat tolerance requires the entry of extracellular Ca2+ into cells across the plasma membrane and the mediation of intracellular CaM.
314. K. Y. Lim et al., “A Genetic Appraisal of a New Synthetic Nicotiana Tabacum (Solanaceae) and the Kostoff Synthetic Tobacco,” American Journal of Botany, vol. 93, no. 6, pp. 875–883, 2006. doi: 10.3732/ajb.93.6.875. Abstract
     Polyploids have significantly influenced angiosperm evolution. Understanding the genetic consequences of polyploidy is advanced by studies on synthetic allopolyploids that mimic natural species. In Nicotiana, Burk (1973) and Kostoff (1938) generated synthetic tobacco (N. tabacum) using the parents ♀N. sylvestris × ♂N. tomentosiformis. We previously reported rapid genetic changes in the Burk material. Kostoff’s material has 24 chromosomes of N. sylvestris origin (S-genome), 24 of N. tomentosiformis origin (T-genome), and a large intergenomic translocation, but not an additive distribution of ribosomal DNA (rDNA) families as expected from the parental contribution. Our new synthetic tobacco lines TR1 and TR2 are chromosomally balanced with no intergenomic translocations and are either sterile or have highly reduced fertility, supporting the nuclear cytoplasmic hypothesis that allopolyploid fertility is enhanced by intergenomic translocations. Two plants of TR1 (TR1-A, TR1-B) have the expected number, structure, and chromosomal distribution of rDNA families, in contrast to Burk’s and Kostoff’s synthetic tobaccos and to synthetic polyploids of Arabidopsis. Perhaps allopolyploids must pass through meiosis before genetic changes involving rDNA become apparent, or the genetic changes may occur stochastically in different synthetic allopolyploids. The lack of fertility in the first generation of our synthetic tobacco lines may have uses in biopharmacy.
315. W. J. S. Lockley, H. H. Rees, and T. W. Goodwin, “Biosynthesis of Steroidal Withanolides in Withania Somnifera,” Phytochemistry, vol. 15, no. 6, pp. 937–939, Jan. 1976. doi: 10.1016/S0031-9422(00)84374-5. Abstract
     Administration of 24-methylene-cholesterol-[28-3H] to Withania somnifera, yielded [3H] radioactivity in the isolated withaferin A and withanolide D, whereas administered 24-(R,S)-methyl-cholesterol-[28-3H] was not incorporated into these compounds. 24-Methylene-cholesterol is, therefore, proposed as a sterol precursor of the withanolides. A novel procedure is described for the isolation of withanolides from W. somnifera. This method in conjunction with an improved procedure for administration of labelled sterols and mevalonolactone produces a greatly increased yield of labelled withanolides.
316. L. R. López-Lefebre, R. M. Rivero, P. C. García, E. Sánchez, J. M. Ruiz, and L. Romero, “Effect of Calcium on Mineral Nutrient Uptake and Growth of Tobacco,” Journal of the Science of Food and Agriculture, vol. 81, no. 14, pp. 1334–1338, 2001. doi: 10.1002/jsfa.948. Abstract
     The aim of this work was to analyse the dynamics of the nutritional state and biomass production in tobacco plants resulting from the application of various calcium levels (Ca1, 1.25 mol m−3 CaCl2; Ca2, 2.50 mol m−3 CaCl2; Ca3, 5 mol m−3 CaCl2) in the culture medium. Tobacco plants were grown under controlled conditions and submitted to regular fertilisation with macro- and micronutrients. The concentrations of Ca, organic N, P, K, Na, Mg, Fe, Mn, Zn, Cu, Cl and B were determined in the roots and leaves. As expected, Ca accumulated progressively with increasing concentration of this element in the culture medium. There was a slight rise in the concentration of organic N but hardly any change in the concentrations of K and Na. In contrast, increasing Ca application caused a gradual decline in P and Mg concentrations. In terms of micronutrient concentrations, Fe, Mn, Zn, Cl and B were influenced positively by Ca treatment. A noteworthy synergetic relationship was found between Ca and B. In contrast, the Cu concentration in the roots declined significantly with increasing Ca application. The effect of Ca on biomass production depended on the organ analysed, since the dry matter content diminished in the roots but was augmented slightly in the leaves. © 2001 Society of Chemical Industry
317. J. H. Loughrin, T. R. Hamilton-Kemp, R. A. Andersen, and D. F. Hildebrand, “Headspace Compounds from Flowers of Nicotiana Tabacum and Related Species,” ACS Publications. American Chemical Society, May-2002. doi: 10.1021/jf00092a027.
318. R. H. Lowe, “Effect of Light on Curing of Burley Tobacco.”
319. W. M. Lush and A. E. Clarke, “Observations of Pollen Tube Growth in Nicotiana Alata and Their Implications for the Mechanism of Self-Incompatibility,” Sexual Plant Reproduction, vol. 10, no. 1, pp. 27–35, Feb. 1997. doi: 10.1007/s004970050064. Abstract
     Style squashes and stylar grafts were used to examine the growth of Nicotiana alata pollen tubes in self-compatible and self-incompatible styles. Compatible tubes typically showed a uniform layer of callose deposition in the walls and in small plugs spaced at regular intervals within the tube. Incompatible tubes were characterised by the variability of callose deposition in the walls and by larger, closer and more irregularly spaced plugs. There was no difference in the growth rate of compatible and incompatible tubes during growth through the stigma, but within the style most compatible tubes grew 20–25 mm day-1 (maximum 30 mm day–1), whereas incompatible tubes grew 1.0–1.5 mm day-1 (maximum 5 mm day–1). Many incompatible tubes continued to grow until flowers senesced, and only a small proportion died as a consequence of tip bursting. Grafting compatibly pollinated styles onto incompatible styles showed that the incompatible reaction could occur in pollen tubes between 2 and 50 mm long, and that inhibition of pollen tube growth occurred in both the upper and lower parts of the transmitting tract. Grafting incompatibly pollinated styles onto compatible styles showed that the incompatible reaction was fully reversible in at least a proportion of the pollen tubes. The findings are not consistent with the cytotoxic model of inhibition of self-pollen tubes in solanaceous plants, which assumes that the incompatible response results from the degradation of a finite amount of rRNA present in the pollen tube. However, if pollen tubes do in fact synthesise rRNA, the findings become consistent with this model.
320. W. M. Lush, A. S. Opat, F. Nie, and A. E. Clarke, “An in Vitro Assay for Assessing the Effects of Growth Factors on Nicotiana Alata Pollen Tubes,” Sexual Plant Reproduction, vol. 10, no. 6, pp. 351–357, Dec. 1997. doi: 10.1007/s004970050109. Abstract
     A new method for assessing the effects of test compounds on Nicotiana alata pollen tubes in culture is described. Pollen tubes grow from a cluster of grains placed beneath a thin layer of gelled medium in which test substances are incorporated and from which evaporation is prevented by a covering layer of oil. Pollen tubes can grow to 8 mm in length in 24 h, which corresponds to about 25% of the maximum growth rate in styles. Growth is non-destructively measured. The developmental stages reached by cultured tubes are similar to those of tubes growing in styles; growth changes from being reserve-dependent to reserve-independent, callose plugs form, and the nucleus of the generative cell divides. Because culture volumes are small (10–20 μl per replicate), the effects of known concentrations of microgram quantities of compounds on the growth of pollen tubes can be tested.
321. D. Lyra, D. Kalivas, and G. Economou, “A Large-Scale Analysis of Soil and Bioclimatic Factors Affecting the Infestation Level of Tobacco (Nicotiana Tabacum L.) by Phelipanche Species,” Crop Protection, vol. 83, pp. 27–36, May 2016. doi: 10.1016/j.cropro.2016.01.008. Abstract
     Soil and bioclimatic factors constitute key factors that influence the interaction between host plants and broomrape. The identification and quantification of the relationship between environmental variables and the level of infestation with Phelipanche aegyptiaca and Phelipanche ramosa was examined for 144 tobacco fields in surveys during two years 2003–2004 in Greece. The distribution of Phelipanche species was mapped using a Geographical Information System to reveal variability in the infestation level between the species, the cultivated types and the sampling areas. To assess the effect of abiotic factors on the level of infestation with Phelipanche species, soil and bioclimatic parameters were taken into account: soil texture, pH, organic matter content (OM), total humidity index (THI) and growing degree days (GDD). Univariate and multivariate statistics were used to examine the influence of the above parameters on Phelipanche intensity. ANOVA showed that the infestation varied significantly between the sampling areas and Phelipanche species, while significant differences were also detected among the regions in terms of the studied abiotic parameters. Correlation analysis demonstrated that the level of Phelipanche infestation correlated negatively with pH, THI and positively with OM. Multiple regression analysis suggested that soil and climatic variables together explained 63% of the infestation variance. Discriminant functions accounted for 90% of the total variation in the dataset. All statistical analyses demonstrated that pH, THI and OM were the most decisive variables for the severity of infestation. The two Phelipanche species were clearly discriminated, with P. ramosa being found at higher and P. aegyptiaca at a lower level of infestation. A parasitism specification was also observed between the two Phelipanche species and tobacco types. The findings provide constructive information for modelling broomrape infestation based on abiotic factors and create a baseline for future monitoring of broomrape distribution.
322. F. D. H. Macdowall, “Organic Requirements for Culture of Excised Roots of Nicotiana Rustica,” Botanical Gazette, vol. 123, no. 3, pp. 180–185, Mar. 1962. doi: 10.1086/336147. Abstract
     1. The organic constituents of White’s medium were tested for their effectiveness in supporting the growth of isolated roots of Nicotiana rustica. 2. Glycine and nicotinic acid were toxic. 3. Sucrose at 2%-4% was optimal for growth on the dry weight basis but the higher concentrations, including 4%, inhibited cell elongation. 4. Pyridoxine and thiamin were required for good growth. 5. A clone has been perpetuated for 5 years in White’s medium without glycine and nicotinic acid, and the probability of its indefinite growth was suggested.
323. S. K. Maheshwari, R. K. Sharma, and S. K. Gangrade, “Response of Ashwagandha (Withania Somnifera) to Organic Manures and Fertilizers in a Shallow Black Soil under Rainfed Condition,” Indian Journal of Agronomy, vol. 45, pp. 214–216, Mar. 2000. Abstract
     In a 2-year field study the highest dry-root yield, maximum net return and higher benefit : cost ratio of ashwagandha [Withania somnifera (L.) Dunal] were recorded by applying 2.5 tonnes/ ha farmyard manure along with 12.5 kg N + 25 kg P2O5/ha. Under rainfed condition in a shallow black soil. Application of 5 t farmyard manure/ha was also found suitable for increasing the dry-root yield. Cultivar ’JA 20’ performed very well compared with ’JA 134’.
324. W. S. Malloch and F. W. Malloch, “Species Crosses in Nicotiana, with Particular Reference to N. LONGIFLORA x N. TABACUM, N. LONGIFLORA x N. SANDERAE, N. TABACUM x N. GLAUCA,” Genetics, vol. 9, no. 3, pp. 261–291, May 1924. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1200821/.
325. S. Mallubhotla et al., “Field Performance, Rapd Analysis and Withanolide Profiling of Micropropagated Plants of Wsr - an Elite Cultivar of Indian Ginseng (Withania Somnifera L. Dunal),” Acta Horticulturae, no. 865, pp. 199–207, Jun. 2010. doi: 10.17660/ActaHortic.2010.865.25.
326. S. Manohar, M. R. Choudhary, B. L. Yadav, S. Dadheech, and S. P. Singh, “Analyzing the Efficacy of Organic and Inorganic Sources of Nitrogen and Phosphorus on Growth of Ashwagandha (Withania Somnifera Dunal.),” Journal of Horticultural Science, vol. 7, no. 2, pp. 161–165, Jan. 2020. https://jhs.iihr.res.in/index.php/jhs/article/view/369. Abstract
     Ashwagandha (Withania somnifera Dunal.) is an important medicinal plant whose roots are prescribed as medicine for several disorders of females, bronchitis, dropsy, stomach problems, lung inflammation, tuberculosis, arthritis, skin diseases and male impotency. The present experiment was designed to work out a suitable dose of organic manures and fertilizers for ashwagandha. Treatments consisted of nitrogenous (N) and phosphatic (P) fertilizers at 20 kg ha-1 and 40 kg ha-1 each, and two levels of farm yard manure (FYM) and vermicompost and combinations thereof, along with control. The treatments were replicated thrice in Randomized Block Design. Results revealed that application of 40 kg ha-1 of N and P each as urea and SSP + 2.5 t ha-1 vermicompost registered significant values for plant height, number of branches per plant, leaf area, yield attributing traits, root (8.60 q ha-1) and seed yield (85.6 kg ha-1) as well as soil physical properties like organic carbon, hydraulic conductivity and water retention at 33 and 1500kpa besides the highest B:C ratio (2.57).
327. S. Martin, D. Saco, and M. Alvarez, “Nitrogen Metabolism in Nicotian a Rustica L. Grown with Molybdenum. II. Flowering Stage,” Communications in Soil Science and Plant Analysis, vol. 26, no. 11-12, pp. 1733–1747, Jun. 1995. doi: 10.1080/00103629509369405. Abstract
     The effect of molybdenum application time on development and nitrogen metabolism in Nicotiana rustica L. was studied. The plants were grown in hydroponically in vermiculite. The control plants received 0.01 ppm Mo, and the treated plants received 1 ppm Mo throughout the experiment (1‐Mo lot) and 1 ppm Mo only until flowering (1‐Mo+C lot). Molybdenum supply accelerated the flowering period in N. rustica, mainly when the supply is only during the vegetative stage. Nitrate reductase activity and root nitrate content increased with molybdenum supply. In contrast, nitrate content in leaves decreased with this molybdenum dose. Both effects are independent of the molybdenum application time. Important differences between molybdenum application time in N. rustica were not observed, except in the root, where the protein content was greater in 1‐Mo+C treatment.
328. J. Martínez-Pacheco, “In Vitro Germination of Tobacco (Nicotiana Tabacum L.) Pollen,” Cultivos Tropicales, vol. 39, no. 1, pp. 102–107, Mar. 2018. http://scielo.sld.cu/scielo.php?script=sci_abstract&pid=S0258-59362018000100013&lng=en&nrm=iso&tlng=es.
329. J. Masaka, N. Chimwanda, I. Chagonda, and M. Chandiposha, “A Comparative Evaluation of the Physical and Chemical Characteristics of Composted Tea Tree (Melaleuca Alternifolia L.) with Pine Bark Growing Media in Tobacco (Nicotiana Tabucum L.) Seedling Production,” Advances in Agriculture, vol. 2016, p. e5650290, Feb. 2016. doi: 10.1155/2016/5650290. Abstract
     The search for suitable organic growing media substitutes for pine bark based media combinations has been prompted by concerns over high costs and lack of availability to smallholder farmers coupled with increasing demand for soilless media in Zimbabwe. A trial was conducted for 12 weeks to investigate the suitability of tea tree compost-based substrates for tobacco seedling production as a substitute of the traditional pine bark growing media. The use of composted tea tree growing media in float trays significantly () increases the bulk density of the media by 23–59% when compared with pine bark media. The cation exchange capacity of the native pine bark growing media was 14 to 95% lower than that for the composted tea tree media. The use of the composted tea as media for tobacco seedling nursery reduced seed germination by 10–37%, seedling stem height by 4–34%, and seedling stem girth by 6–175%. While the nutrient holding and supplying potential of the growing media in seedling production is important for normal seedling growth, its effect on seedling growth vigor is less important than that exerted by the presence of suitable physical conditions in the media.
330. K. Mather and A. Vines, “The Inheritance of Height and Flowering Time in a Cross of Nicotiana Rustica.,” The inheritance of height and flowering time in a cross of Nicotiana rustica., 1952. https://www.cabdirect.org/cabdirect/abstract/19541603509. Abstract
     The mode of inheritance of the quantitative characters height and flowering time in Nicotiana rustica was studied by raising F1 F2, F3 and F4 families, sib-cross families and first and second generation back-cross families of a selected cross. Scale transformations were not made, though the direct readings were not fully adequate. Variation was partitioned into components for genetic effect,...
331. E. Matusiewicz, “The influence of thinning the flowers of Nicotiana rustica on the yield and oil content of the seeds.,” Rocznik nauk rolniczych, vol. 81, pp. 805–17, 1960. https://www.cabdirect.org/cabdirect/abstract/19620307024. Abstract
     Four treatments were compared, namely, removing all flowers and leaving only 10 leaves, reducing the number of flowers to leave 50 seed capsules, reducing the number of flowers to leave 100 capsules and a control which received no treatment. The greatest yield of leaves was obtained where only 10 leaves were left. The yield of leaves and their content of nicotine decreased as the number of seed...
332. E. Matusiewicz, “Obtaining nicotine from Nicotiana rustica.,” Tyton, Warszawa, vol. 6, no. 9-10, pp. 6–7, 1950. https://www.cabdirect.org/cabdirect/abstract/19511602903. Abstract
     In two years’ experiments with seven varieties of N. rustica, chosen beforehand for their high content of nicotine, the highest yield of nicotine was obtained from Pomorska Selection 4 and the highest average yield of nicotine for the two years from the hybrid Czerbl x Selvaggio 85/10.
333. E. Matusiewicz, “Studies on the nutrient requirements of Nicotiana rustica, with special attention to yield and seed quality.,” Rocznik nauk rolniczych, vol. 81, pp. 819–28, 1960. https://www.cabdirect.org/cabdirect/abstract/19620307001. Abstract
     During early growth adequate N and P were important, whereas K was of minor importance; during flowering, however, N and K were needed in larger amounts and the P requirement declined. Leaf size and yield depended primarily on N and to a lesser degree on P. P deficiency caused an increase in the length of the leaf stalk. The number of seed capsules per plant and the yield of seeds per ha....
334. A. Mazeed, P. Maurya, D. Kumar, and P. Suryavanshi, “The Enhancement of Root Yield and Quality of Ashwagandha [Withania Somnifera (L.) Dunal] by Weeds Leaves Extracts,” Indian Journal of Weed Science, vol. 54, no. 1, pp. 81–86, 2022. doi: 10.5958/0974-8164.2022.00014.4. Abstract
     Ashwagandha, having multiple therapeutic uses, is a highly valuable medicinal plant for pharmaceutical industry. In order to meet the industrial demand, both quality as well as yield of ashwagandha needs to be improved agronomically. In this study, effect of weed leaves extracts (WLE) as bio-stimulants to improve yield and quality of ashwagandha roots were studied in pot experiments during 2020-21. The treatments consisted of combinations of four commercial preparations with microorganisms (Pusa zinc solubilizing biofertilizer, Pusa Azotobacter liquid biofertilizer, Pusa PSB liquid biofertilizer, Pusa Potash solubilizing liquid biofertilizer) with four weeds [Cyperus rotundus L., Amaranthus viridis L., Echinochloa colona (L.) Link, Digera arvensis Forsk.] leaves extracts. The treated plants exhibited stimulatory responses in growth and physiology, leading to enhanced dry root yield of ashwagandha compared to control. Yield enhancing effects of different treatments, when used separately, without combination was the highest in case of Amaranthus viridis WLE, followed by Digera arvensis WLE and PSB solubilizing biofertilizer, however co-application resulted in synergistic effect. Among different combination of treatments, Pusa PSB liquid biofertilizer + Amaranthus viridis WLE recorded the highest whole ashwagandha plant dry matter production (157.3 g/plant), root fresh weight per plant (65.0 g) and root dry weight (23.0 g). Different bioactive compounds in ashwagandha roots (withanoloides A, withanosides IV and withanone) were also enhanced with this treatment indicating the potentiality of weed leaves extracts as biostimulants, with a possibility to use as a novel eco-friendly approach for enhancing root yield and quality of ashwagandha.
335. K. L. McCorkle, K. Drake-Stowe, R. S. Lewis, and D. Shew, “Characterization of Phytophthora Nicotianae Resistance Conferred by the Introgressed Nicotiana Rustica Region, Wz, in Flue-Cured Tobacco,” Plant Disease, vol. 102, no. 2, pp. 309–317, Feb. 2018. doi: 10.1094/PDIS-03-17-0339-RE. Abstract
     Black shank, caused by Phytophthora nicotianae, is one of the most important diseases affecting tobacco worldwide and is primarily managed through use of host resistance. An additional source of resistance to P. nicotianae, designated as Wz, has been introgressed into Nicotiana tabacum from N. rustica. The Wz gene region confers high levels of resistance to all races, but has not been characterized. Our study found Wz-mediated resistance is most highly expressed in the roots, with only a slight reduction in stem-lesion size in Wz genotypes compared with susceptible controls. No substantial relationships were observed between initial inoculum levels and disease development on Wz genotypes, which is generally consistent with qualitative or complete resistance. Isolates of P. nicotianae adapted for five host generations on plants with the Wz gene caused higher disease severity than isolates adapted on Wz plants for only one host generation. Wz-adapted isolates did not exhibit increased aggressiveness on genotypes with other sources of partial resistance, suggesting pathogen adaptation was specific to the Wz gene. To reduce potential for pathogen population shifts with virulence on Wz genotypes, Wz should be combined with other resistance sources and rotation of varying black shank resistance mechanisms is also recommended.
336. L. Meena et al., “Ashwagandha (Withania Somnifera L.),” in Medicinal Plants in India: Importance and Cultivation, 2020, pp. 81–94. Abstract
     Ashwagandha (Withania somnifera L.) is an annual medicinal plant belongs to family Solanaceae and known as winter cherry. Among the various medicinal plants, Withania somnifera (L.) Dunal (Winter cherry, Ashwagandha or Asgandh) is an important medicinal plant and its use in Ayurveda and Unani medicine extends back over 3000 to 4000 years.The Sanskrit name “ashva” meaning horse and “gandha” meaning smelling was given to this plant due to the smell of the roots resembling a sweating horse. Ashwagandha was first mentioned by sage Punarvasu Atreya over 4000 years ago (Rajeswara Rao et al., 2012)
337. F. Mehmood et al., “Plastid Genomics of Nicotiana (Solanaceae): Insights into Molecular Evolution, Positive Selection and the Origin of the Maternal Genome of Aztec Tobacco (Nicotiana Rustica),” PeerJ, vol. 8, p. e9552, Jul. 2020. doi: 10.7717/peerj.9552. Abstract
     Species of the genus Nicotiana (Solanaceae), commonly referred to as tobacco plants, are often cultivated as non-food crops and garden ornamentals. In addition to the worldwide production of tobacco leaves, they are also used as evolutionary model systems due to their complex development history tangled by polyploidy and hybridization. Here, we assembled the plastid genomes of five tobacco species: N. knightiana, N. rustica, N. paniculata, N. obtusifolia and N. glauca. De novo assembled tobacco plastid genomes had the typical quadripartite structure, consisting of a pair of inverted repeat (IR) regions (25,323–25,369 bp each) separated by a large single-copy (LSC) region (86,510–86,716 bp) and a small single-copy (SSC) region (18,441–18,555 bp). Comparative analyses of Nicotiana plastid genomes with currently available Solanaceae genome sequences showed similar GC and gene content, codon usage, simple sequence and oligonucleotide repeats, RNA editing sites, and substitutions. We identified 20 highly polymorphic regions, mostly belonging to intergenic spacer regions (IGS), which could be suitable for the development of robust and cost-effective markers for inferring the phylogeny of the genus Nicotiana and family Solanaceae. Our comparative plastid genome analysis revealed that the maternal parent of the tetraploid N. rustica was the common ancestor of N. paniculata and N. knightiana, and the later species is more closely related to N. rustica. Relaxed molecular clock analyses estimated the speciation event between N. rustica and N. knightiana appeared 0.56 Ma (HPD 0.65–0.46). Biogeographical analysis supported a south-to-north range expansion and diversification for N. rustica and related species, where N. undulata and N. paniculata evolved in North/Central Peru, while N. rustica developed in Southern Peru and separated from N. knightiana, which adapted to the Southern coastal climatic regimes. We further inspected selective pressure on protein-coding genes among tobacco species to determine if this adaptation process affected the evolution of plastid genes. These analyses indicate that four genes involved in different plastid functions, including DNA replication (rpoA) and photosynthesis (atpB, ndhD and ndhF), came under positive selective pressure as a result of specific environmental conditions. Genetic mutations in these genes might have contributed to better survival and superior adaptations during the evolutionary history of tobacco species.
338. M. Mench, J. Tancogne, A. Gomez, and C. Juste, “Cadmium Bioavailability to Nicotiana Tabacum L., Nicotiana Rustica L., and Zea Mays L. Grown in Soil Amended or Not Amended with Cadmium Nitrate,” Biology and Fertility of Soils, vol. 8, no. 1, pp. 48–53, Jul. 1989. doi: 10.1007/BF00260515. Abstract
     Mature (flowering) tobacco (Nicotiana tabacum cv. PBD6, Nicotiana rustica cv. Brasilia) and maize (Zea mays cv. INRA 260) plants were grown in an acid sandy-clay soil, enriched to 5.4 mg Cd kg−1 dry weight soil with cadmium nitrate. The plants were grown in containers in the open air. No visible symptoms of Cd toxicity developed on plant shoots over the 2-month growing period. Dry-matter yields showed that while the Nicotiana spp. were unaffected by the Cd application the yield of Z. mays decreased by 21%. Cd accumulation and distribution in leaves, stems and roots were examined. In the control treatment (0.44 mg Cd kg−1 dry weight soil), plant Cd levels ranged from 0.4 to 6.8 mg kg−1 dry weight depending on plant species and plant parts. Soil Cd enrichment invariably increased the Cd concentrations in plant parts, which varied from 10.1 to 164 mg kg−1 dry weight. The maximum Cd concentrations occurred in the leaves of N. tabacum. In N. rustica 75% of the total Cd taken up by the plant was transported to the leaves, and 81% for N. tabacum irrespective of the Cd level in the soil. In contrast, the Cd concentrations in maize roots were almost five times higher than those in the leaves. More than 50% of the total Cd taken up by maize was retained in the roots at both soil Cd levels. The Cd level in N. tabacum leaves was 1.5 and 2 times higher at the low and high Cd soil level, respectively, than that in N. rustica leaves, but no significant difference was found in root Cd concentrations between the two Nicotiana spp.
339. M. Mench and E. Martin, “Mobilization of Cadmium and Other Metals from Two Soils by Root Exudates of Zea Mays L., Nicotiana Tabacum L. and Nicotiana Rustica L.,” Plant and Soil, vol. 132, no. 2, pp. 187–196, Apr. 1991. doi: 10.1007/BF00010399. Abstract
     Soluble root exudates were collected from three plants (Nicotiana tabacum L., Nicotiana rustica L. and Zea mays L.), grown under axenic and hydroponic conditions, in order to study their metal-solubilizing ability for Cd and other cations (Cu, Fe, Mn, Ni, Zn). Nicotiana spp. and Zea mays L. root exudates differed markedly in C/N ratio, sugars vs. amino acids ratio and organic acids content. Metals from two soils were extracted with either root exudate solutions, containing equal amounts of organic carbon, or distilled water as control. In the presence or absence of root exudates, the solubility of Fe and Mn was much higher than of the four other metals tested. Root exudates increased the solubilities of Mn and Cu, whereas those of Ni and Zn were not affected. Root exudates of Nicotiana spp. enhanced the solubility of Cd. The extent of Cd extraction by root exudates (N. tabacum L. N. rustica L. Zea mays L.) was similar to the order of Cd bioavailability to these three plants when grown on soil. An increase in Cd solubility in the rhizosphere of apical root zones due to root exudates is likely to be an important cause of the relatively high Cd accumulation in Nicotiana spp.
340. F. Meriama et al., “Experimental Determination and Modeling of the Moisture-Sorption Isotherms and Isosteric Heat of Tobacco Leaves,” Instrumentation Mesure Métrologie, vol. 20, no. 5, pp. 269–277, Oct. 2021. doi: 10.18280/i2m.200504. Abstract
     During a forced convection sun drying and storage operation, the equilibrium water content of a product to be dried is critical. These figures are frequently derived using isothermal sorption curves. The calculation of isotherms is a necessary step in determining the distribution and intensity of water connections in products. for that this paper concentrates on the experimental determination of the adsorption-desorption isotherms for various temperatures (40, 50, and 60℃) of the Nicotiana Tabacum L plants. From which we had established the relationship between the water activity and the water content in the product. However, the aforementioned determination was carried out by the static gravimetric method. Eight saturated salt solutions have been utilized in applications such as (KOH, KCl, MgCl2, MgNO3, K2CO3, BaCl2, K2SO4, and NaCl). Hygroscopic equilibrium was completed after 13 days for temperature 40℃, 11 days for 50℃, and 9 days for 60℃. The overall experimental sorption curves are summarized by six models (HENDERSON, modified HALSEY, OSWIN, GAB, modified BET, and PELEG). The sorption isotherms built using the Clausius–Clapeyron equation were used to determine the net isosteric temperatures of desorption and adsorption of Nicotiana Tabacum L. The results for the adsorption-desorption isotherms found are type III according to IUAPC. Following the smoothing of the experimental results by different used models, it was found that the models of GAB and Peleg allow having the lowest mean relative errors and correlation coefficient.
341. B. A. Mir, S. Koul, A. Kumar, M. K. Kaul, A. S. Soodan, and S. N. Raina, “Assessment and Characterization of Genetic Diversity in Withania Somnifera (L.) Dunal Using RAPD and AFLP Markers,” African Journal of Biotechnology, vol. 10, no. 66, pp. 14746–14756, 2011. doi: 10.4314/ajb.v10i66. Abstract
     Genetic diversity of 23 accessions of Withania somnifera collected from different geographical regions of India was estimated by employing Random Amplification of Polymorphic DNA (RAPD) and Amplified Fragment Length Polymorphism (AFLP) markers. Eighteen RAPD primers and six AFLP primer combinations revealed 37.82 and 43.94% polymorphism, respectively, among 163 and 286 genetic loci amplified. The AFLP assay revealed higher levels of polymorphism among the tested W. somnifera accessions compared to the RAPD. Mean genetic diversity based on Shannon index ranged from 1.33 (RAPD) to 5.13 (AFLP). Unweighted pair group method based on arithmetic average (UPGMA) analysis was performed on Jaccard’s similarity coefficient matrix. The matrix reveals, two main clusters, wild accessions formed one cluster and the cultivated accessions formed the other. The cultivated accessions are well separated from the wild ones at a low similarity value of 0.3, indicating that cultivated and wild accessions are highly distinct. Morphologically cultivated accessions were also quite distinct from the wild ones and the cluster analysis of RAPD and AFLP fingerprints clearly discriminated the five cultivated accessions of W. somnifera. A strong correlation was observed between morphology and molecular marker systems. Identification of specific markers to wild as well as cultivated accessions is yet another important finding in the present study. Such genetic diversity is useful in facilitating the development of large number of new varieties through hybridization, transfer of useful genes, thus maximizing the use of such available germplasms as genetic resource materials for breeders. The present input, first of its kind in Ashwagandha, will thus assist the marker assisted crop improvement programme.Key words: Withania somnifera, genetic diversity, RAPD, AFLP, polymorphism, Shannon index.
342. B. A. Mir et al., “Cold Stress Affects Antioxidative Response and Accumulation of Medicinally Important Withanolides in Withania Somnifera (L.) Dunal,” Industrial Crops and Products, vol. 74, pp. 1008–1016, Nov. 2015. doi: 10.1016/j.indcrop.2015.06.012. Abstract
     Withania somnifera (L.) Dunal (Indian ginseng) is a high value medicinal plant. It synthesizes a large array of biologically active withanolides. In this study, two month old seedlings of AGB002 (wild genotype) and AGB025 (cultivated genotype) of W. somnifera were subjected to cold stress (4°C) under controlled environment. Plants were analyzed for three medicinally important secondary metabolites (withanolide A, withanone and withaferin A), lipid peroxidation (MDA), cell injury, superoxide radical (O2−) accumulation and anti-oxidative enzymes activities such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR). Increases in the titers of superoxide anion and MDA were observed from day 1 to day 7 in both genotypes, although the increase on the first day of exposure was significantly higher. Enzymatic activities of SOD, CAT, APX and GR also showed an increasing trend in both genotypes and reached a maximum on day 7 of the cold temperature exposure; however, this increase was higher in AGB002 than AGB025. Withanolide A (WS-1) in the roots of both genotypes significantly decreased on the first day of cold exposure and then showed a recovery until day 7. WS-1 was not detected in the leaves of either genotype. Withanone (WS-2) content in the leaves also decreased towards the end of the cold period in both genotypes. Cold stress also elicited the accumulation of WS-2 in AGB025, but was not detectable in control seedlings. At maturity, WS-2 was also detected in control plants. Furthermore, a significant increase in the leaf withaferin A (WS-3) content was recorded from day 1 to day 7 of the cold exposure in both the genotypes, suggesting the possible involvement of withanolides in cold-protection. AGB002 showed comparatively higher accumulation of antioxidant enzymes and selected marker withanolides than AGB025, indicating that AGB002 is better adapted to cold than AGB025. It could be inferred from these observations that cold stress induces bioactive withanolide accumulation in W. somnifera as a mechanism for scavenging reactive oxygen species (ROS). These studies also provide an impetus for enhancing the withanolide accumulation in W. somnifera using controlled environment technology.
343. B. A. Mir, J. Khazir, K. R. Hakeem, S. Koul, and D. A. Cowan, “Enhanced Production of Withaferin-A in Shoot Cultures of Withania Somnifera (L) Dunal,” Journal of Plant Biochemistry and Biotechnology, vol. 23, no. 4, pp. 430–434, Oct. 2014. doi: 10.1007/s13562-014-0264-8. Abstract
     Withania somnifera (L) Dunal, commonly known as ashwagandha or Indian ginseng, is the source of large number of pharmacologically active withanolides. Withaferin-A (WS-3), a major withanolide of W. somnifera, has been proven to be an effective anti-cancer molecule. In this study, a liquid culture system for shoot proliferation, biomass accumulation and withaferin-A production of an elite accession (AGB002) of W. somnifera was investigated. The nodal explants cultured on Murashige and Skoog (MS) semi-solid medium supplemented with various concentrations of 6-benzyl adenine (BA) and Kinetin (Kn) elicited varied responses. The highest number of regenerated shoots per ex-plant (35\,± 3.25) and the maximum average shoot length (5.0\,± 0.25 cm) were recorded on MS medium supplemented with BA (5.0 μM). The shoots were further proliferated in half and full strength MS liquid medium supplemented with the same concentration BA. It was interesting to note that shoots cultured on MS half strength liquid medium fortified with 4 gL-1 FW (fresh weight) shoot inoculum mass derived from 5 week old nodal explants of W. somnifera showed highest accumulation of biomass and withaferin A content in 5 weeks. Withaferin A was produced in relatively high amounts (1.30 % and 1.10 % DW) in shoots cultured in half and full strength MS liquid media respectively as compared to natural field grown plants (0.85 % DW). A considerable amount of the withaferin A was also excreted in the culture medium. Successful proliferation of shoots in liquid medium and the synthesis of withaferin A in vitro opens new avenues for bioreactor scale-up and the large-scale production of the compound.
344. B. A. Mir, S. Koul, and A. S. Soodan, “Reproductive Biology of Withania Ashwagandha Sp. Novo (Solanaceae),” Industrial Crops and Products, vol. 45, pp. 442–446, Feb. 2013. doi: 10.1016/j.indcrop.2012.12.023. Abstract
     Withania ashwagandha Kaul (Solanaceae) is an annual plant species of immense medicinal importance. It is a repository of a large number of pharmacologically active secondary metabolites known as withanolides. Evidence for the delimitation of the species from W. somnifera has been provided by our group using multidisciplinary approaches. Knowledge of reproductive biology of medicinal plants is crucial for improvement, effective conservation and management plans to evolve genetically superior varieties. The present paper reports our findings on the floral biology, pollination behavior and breeding system of W. ashwagandha in natural populations grown at our experimental field under near natural conditions. Flowering (peak) takes place during April–July and anthesis occurs between 08:00 and 11:00h. The period of stigma receptivity coincides with anther dehiscence. Fruit set on pollination treatments ranged from 90.8% (passive autogamy), 72% (assisted autogamy), 30.30% (xenogamy), and 56.50% (geitonogamy) through 50.40% (open pollination). Xenogamy brings about very low fruit set, seed-set and seed germination percentages. It is inferred that W. ashwagandha is predominantly an autogamous and self-compatible species. Self-compatibility is mainly accomplished due to close proximity of stigma and anthers. This work is the first report on the reproductive biology of W. ashwagandha and will be useful for conservation and development of improved varieties of this multipurpose herb.
345. M. H. Mirjalili, S. M. Fakhr-Tabatabaei, H. Alizadeh, A. Ghassempour, and F. Mirzajani, “Genetic and Withaferin A Analysis of Iranian Natural Populations of Withania Somnifera and W. Coagulans by RAPD and HPTLC,” Natural Product Communications, vol. 4, no. 3, p. 1934578X0900400307, Mar. 2009. doi: 10.1177/1934578X0900400307. Abstract
     For successful conservation and breeding of a medicinal species, it is important to evaluate its genetic diversity as well as its content of phytochemical compounds. The aim of the present study was to investigate the genetic variation of Iranian natural populations of W. somnifera and W. coagulans, using the RAPD (random amplified polymorphic DNA) markers, and their withaferin A content. Using 16 RAPD primers, a total of 282 RAPD bands were achieved. The highest and lowest percentages of polymorphism were observed with primers OPAD-15 (100.0%) and OPC-06 (75.0%), respectively. Cluster analysis of the genotypes was performed based on data from polymorphic RAPD bands, using Dice’s similarity coefficient and the UPGMA clustering method. Variations in the RAPD results were found to reflect geographical distribution and genetic factors of the plant populations. The HPTLC analysis of the studied samples revealed the presence of withaferin A in W. coagulans and W. somnifera. Moreover, the concentration of withaferin A had a range from 2.2 to 32.5 ?g/g DW and was higher in the aerial part than in the root in all used samples. The results of the present study show that there is a high level of variation in the Iranian natural population of Withania, which is significant for conservation and breeding programs to improve production of withaferin A.
346. B. Mishra and N. Singh Sangwan, “Amelioration of Cadmium Stress in Withania Somnifera by ROS Management: Active Participation of Primary and Secondary Metabolism,” Plant Growth Regulation, vol. 87, no. 3, pp. 403–412, Apr. 2019. doi: 10.1007/s10725-019-00480-8. Abstract
     Cadmium (Cd) is considered as a non-essential heavy metal with substantial toxicity on environment. Withania somnifera, a reputed therapeutic herb exhibits vast pharmacological activities due to the presence of steroidal lactones-withanolides. The present study deals with reactive oxygen species (ROS) management through primary and secondary metabolism as adaptive response, on exogenous Cd exposure of W. somnifera. Increased invertase enzyme activity resulted in higher reducing sugars in plant under Cd stress to provide additional carbon source and NADH or NADPH. Higher activities of proline metabolic pathway enzymes such as ornithine aminotransferase (OAT) and pyrroline-5-corboxylate synthase (P5CS) resulted in elevated proline accumulation. The considerable participation of phenylpropanoid metabolism also found dominantly upregulated. Increased enzyme activities of phenylalanine ammonia lyase (PAL), shikimate dehydrogenase (SKDH), glucose-6-phosphate dehydrogenase (G6PDH), and cinnamyl alcohol dehydrogenase (CADH) resulted in phenolics and flavonoids accumulation under higher Cd stress. Upregulation of glutathione-S-transferase (GST) activity conferred its role in Cd chelation. In addition, glutamate oxaloacetate transaminase (GOT), amyl esterase (EST) and diaphorase (DIA) established their participation in Cd tolerance mechanism. Thus, present study elucidated major role of proline and phenylpropanoid metabolism in providing the osmoticum and antioxidants to the plants under Cd stress. The information may be helpful in developing stress resistant plants by targeting these pathways using conventional and molecular approaches.
347. A. Mishra et al., “Bacterial Endophytes Modulates the Withanolide Biosynthetic Pathway and Physiological Performance in Withania Somnifera under Biotic Stress,” Microbiological Research, vol. 212–213, pp. 17–28, Jul. 2018. doi: 10.1016/j.micres.2018.04.006. Abstract
     Despite the vast exploration of endophytic microbes for growth enhancement in various crops, knowledge about their impact on the production of therapeutically important secondary metabolites is scarce. In the current investigation, chitinolytic bacterial endophytes were isolated from selected medicinal plants and assessed for their mycolytic as well as plant growth promoting potentials. Among them the two most efficient bacterial endophytes namely Bacillus amyloliquefaciens (MPE20) and Pseudomonas fluorescens (MPE115) individually as well as in combination were able to modulate withanolide biosynthetic pathway and tolerance against Alternaria alternata in Withania somnifera. Interestingly, the expression level of withanolide biosynthetic pathway genes (3-hydroxy-3-methylglutaryl co-enzyme A reductase, 1-deoxy-D-xylulose-5-phosphate reductase, farnesyl di-phosphate synthase, squalene synthase, cytochrome p450, sterol desaturase, sterol Δ-7 reductase and sterol glycosyl transferases) were upregulated in plants treated with the microbial consortium under A. alternata stress. In addition, application of microbes not only augmented withaferin A, withanolide A and withanolide B content (1.52–1.96, 3.32–5.96 and 12.49–21.47 fold, respectively) during A. alternata pathogenicity but also strengthened host resistance via improvement in the photochemical efficiency, normalizing the oxidized and non-oxidized fraction, accelerating photochemical and non-photochemical quantum yield, and electron transport rate. Moreover, reduction in the passively dissipated energy of PSI and PSII in microbial combination treated plants corroborate well with the above findings. Altogether, the above finding highlights novel insights into the underlying mechanisms in application of endophytes and emphasizes their capability to accelerate biosynthesis of withanolides in W. somnifera under biotic stress caused by A. alternata.
348. B. Mishra, R. S. Sangwan, S. Mishra, J. S. Jadaun, F. Sabir, and N. S. Sangwan, “Effect of Cadmium Stress on Inductive Enzymatic and Nonenzymatic Responses of ROS and Sugar Metabolism in Multiple Shoot Cultures of Ashwagandha (Withania Somnifera Dunal),” Protoplasma, vol. 251, no. 5, pp. 1031–1045, Sep. 2014. doi: 10.1007/s00709-014-0613-4. Abstract
     Withania somnifera is one of the most important medicinal plant and is credited with various pharmacological activities. In this study, in vitro multiple shoot cultures were exposed to different concentrations (5–300 μM) of cadmium (Cd) as cadmium sulphate to explore its ability to accumulate the heavy metal ion and its impact on the metabolic status and adaptive responses. The results showed that supplemental exposure to Cd interfered with N, P, and K uptake creating N, P, and K deficiency at higher doses of Cd that also caused stunting of growth, chlorosis, and necrosis. The study showed that in vitro shoots could markedly accumulate Cd in a concentration-dependent manner. Enzymatic activities and isozymic pattern of catalase, ascorbate peroxidase, guaiacol peroxidase, peroxidase, glutathione-S-transferase, glutathione peroxidase, monodehydroascorbate reductase, and dehydroascorbate reductase were altered substantially under Cd exposure. Sugar metabolism was also markedly modulated under Cd stress. Various other parameters including contents of photosynthetic pigments, phenolics, tocopherol, flavonoids, reduced glutathione, nonprotein thiol, ascorbate, and proline displayed major inductive responses reflecting their protective role. The results showed that interplay of enzymatic as well as nonenzymatic responses constituted a system endeavor of tolerance of Cd accumulation and an efficient scavenging strategy of its stress implications.
349. A. Mishra et al., “Endophyte-Mediated Modulation of Defense-Related Genes and Systemic Resistance in Withania Somnifera (L.) Dunal under Alternaria Alternata Stress,” Applied and Environmental Microbiology, vol. 84, no. 8, pp. e02845–17, Apr. 2018. doi: 10.1128/AEM.02845-17. Abstract
     Endophytes have been explored and found to perform an important role in plant health. However, their effects on the host physiological function and disease management remain elusive. The present study aimed to assess the potential effects of endophytes, singly as well as in combination, in Withania somnifera (L.) Dunal, on various physiological parameters and systemic defense mechanisms against Alternaria alternata. Seeds primed with the endophytic bacteria Bacillus amyloliquefaciens and Pseudomonas fluorescens individually and in combination demonstrated an enhanced vigor index and germination rate. Interestingly, plants treated with the two-microbe combination showed the lowest plant mortality rate (28%) under A. alternata stress. Physiological profiling of treated plants showed improved photosynthesis, respiration, transpiration, and stomatal conductance under pathogenic stress. Additionally, these endophytes not only augmented defense enzymes and antioxidant activity in treated plants but also enhanced the expression of salicylic acid- and jasmonic acid-responsive genes in the stressed plants. Reductions in reactive oxygen species (ROS) and reactive nitrogen species (RNS) along with enhanced callose deposition in host plant leaves corroborated well with the above findings. Altogether, the study provides novel insights into the underlying mechanisms behind the tripartite interaction of endophyte-A. alternata-W. somnifera and underscores their ability to boost plant health under pathogen stress. IMPORTANCE W. somnifera is well known for producing several medicinally important secondary metabolites. These secondary metabolites are required by various pharmaceutical sectors to produce life-saving drugs. However, the cultivation of W. somnifera faces severe challenge from leaf spot disease caused by A. alternata. To keep pace with the rising demand for this plant and considering its capacity for cultivation under field conditions, the present study was undertaken to develop approaches to enhance production of W. somnifera through intervention using endophytes. Application of bacterial endophytes not only suppresses the pathogenicity of A. alternata but also mitigates excessive ROS/RNS generation via enhanced physiological processes and antioxidant machinery. Expression profiling of plant defense-related genes further validates the efficacy of bacterial endophytes against leaf spot disease.
350. S. Mishra, S. Bansal, R. S. Sangwan, and N. S. Sangwan, “Genotype Independent and Efficient Agrobacterium-Mediated Genetic Transformation of the Medicinal Plant Withania Somnifera Dunal,” Journal of Plant Biochemistry and Biotechnology, vol. 25, no. 2, pp. 191–198, Apr. 2016. doi: 10.1007/s13562-015-0324-8. Abstract
     Withania somnifera one of the most reputed Indian medicinal plant has been extensively used in traditional and modern medicines as active constituents. A high frequency genotype and chemotype independent Agrobacterium-mediated transformation protocol has been developed for W. somnifera by optimizing several factors which influence T-DNA delivery. Leaf and node explants of Withania chemotype was transformed with A. tumefaciens strain GV3101 harboring pIG121Hm plasmid containing the gusA gene encoding β-glucuronidase (GUS) as a reporter gene and the hptII and the nptII gene as selection markers. Various factors affecting transformation efficiency were optimized; as 2 days preconditioning of explants on MS basal supplemented with TDZ 1 μM, Agrobacterium density at OD600 0.4 with inclusion of 100 μM acetosyringone (As) for 20 min co-inoculation duration with 48 h of co-cultivation period at 22 °C using node explants was found optimal to improved the number of GUS foci per responding explant from 36\,± 13.2 to 277.6\,± 22.0, as determined by histochemical GUS assay. The PCR and Southern blot results showed the genomic integration of transgene in Withania genome. On average basis 11 T0 transgenic plants were generated from 100 co-cultivated node explants, representing 10.6 % transformation frequency. Our results demonstrate high frequency, efficient and rapid transformation system for further genetic manipulation in Withania for producing engineered transgenic Withania shoots within very short duration of 3 months.
351. S. Mishra et al., “RNAi and Homologous Over-Expression Based Functional Approaches Reveal Triterpenoid Synthase Gene-Cycloartenol Synthase Is Involved in Downstream Withanolide Biosynthesis in Withania Somnifera,” PLOS ONE, vol. 11, no. 2, p. e0149691, Feb. 2016. doi: 10.1371/journal.pone.0149691. Abstract
     Withania somnifera Dunal, is one of the most commonly used medicinal plant in Ayurvedic and indigenous medicine traditionally owing to its therapeutic potential, because of major chemical constituents, withanolides. Withanolide biosynthesis requires the activities of several enzymes in vivo. Cycloartenol synthase (CAS) is an important enzyme in the withanolide biosynthetic pathway, catalyzing cyclization of 2, 3 oxidosqualene into cycloartenol. In the present study, we have cloned full-length WsCAS from Withania somnifera by homology-based PCR method. For gene function investigation, we constructed three RNAi gene-silencing constructs in backbone of RNAi vector pGSA and a full-length over-expression construct. These constructs were transformed in Agrobacterium strain GV3101 for plant transformation in W. somnifera. Molecular and metabolite analysis was performed in putative Withania transformants. The PCR and Southern blot results showed the genomic integration of these RNAi and overexpression construct(s) in Withania genome. The qRT-PCR analysis showed that the expression of WsCAS gene was considerably downregulated in stable transgenic silenced Withania lines compared with the non-transformed control and HPLC analysis showed that withanolide content was greatly reduced in silenced lines. Transgenic plants over expressing CAS gene displayed enhanced level of CAS transcript and withanolide content compared to non-transformed controls. This work is the first full proof report of functional validation of any metabolic pathway gene in W. somnifera at whole plant level as per our knowledge and it will be further useful to understand the regulatory role of different genes involved in the biosynthesis of withanolides.
352. B. Mishra, S. Chand, and N. Singh Sangwan, “ROS Management Is Mediated by Ascorbate-Glutathione-α-Tocopherol Triad in Co-Ordination with Secondary Metabolic Pathway under Cadmium Stress in Withania Somnifera,” Plant Physiology and Biochemistry, vol. 139, pp. 620–629, Jun. 2019. doi: 10.1016/j.plaphy.2019.03.040. Abstract
     Being static, plants are frequently exposed to various essential and non-essential heavy metals from the surroundings. This exposure results in considerable ROS generation leading to oxidative stress, the primary response of the plants under heavy metal stress. Withania somnifera is a reputed Indian medicinal plant in Ayurveda, having various pharmacological activities due to the presence of withanolides. The present study deals with the understanding endurance of oxidative stress caused by heavy metal exposure and its management through antioxidant partners in synchronization with secondary metabolites in W. somnifera. The quantitative assessment of enzymatic/non-enzymatic antioxidants revealed significant participation of ascorbate-glutathione-α-tocopherol triad in ROS management. Higher activities of glutathione reductase (GR), monodehydroascorbate reductase (MDHAR) and dehydroascorbate reductase (DHAR) resulted in glutathione and ascorbate accumulation. In addition, superoxide dismutase (SOD), glutathione peroxidase (GPX) and peroxidase (POD) were contributed considerably in ROS homeostasis maintenance. In-situ localization and assays related to ROS generation/scavenging revealed key management of ROS status under Cd stress. Higher antioxidative and reducing power activity attributed to the tolerance capability to the plant. Increased expression of withanolide biosynthetic pathway genes such as WsHMGR, WsDXS, WsDXR and WsCAS correlated with enhanced withanolides. The present study indicated the crucial role of the ascorbate-glutathione-α-tocopherol triad in co-ordination with withanolide biosynthesis in affording the oxidative stress, possibly through a cross-talk between the antioxidant machinery and secondary metabolite biosynthesis. The knowledge may be useful in providing the guidelines for developing abiotic stress resistance in plants using conventional and molecular approaches.
353. L. Misra, P. Mishra, A. Pandey, R. S. Sangwan, and N. S. Sangwan, “1,4-Dioxane and Ergosterol Derivatives from Withania Somnifera Roots,” Journal of Asian Natural Products Research, vol. 14, no. 1, pp. 39–45, Jan. 2012. doi: 10.1080/10286020.2011.622719. Abstract
     The chemical investigation on the n-hexane extract of Withania somnifera roots has yielded octacosane, oleic and stearic fatty acids, stigmasterone, stigmasterol, sitostanone, oleanolic acid along with the ergosterol and 1,4-dioxane derivatives as new compounds. The isolation of alkenyl-1,4-dioxane compound is rare, whereas the ergosterol derivative may have biogenetic significance in the lactone formation in the E ring of withanolides. The presence of a 1,4-dioxane derivative in the nonpolar extract of roots assumes importance as this type of compound has not been reported earlier from W. somnifera. The structures of new compounds were elucidated by spectroscopic methods and chemical transformations.
354. L. Misra, P. Lal, R. S. Sangwan, N. S. Sangwan, G. C. Uniyal, and R. Tuli, “Unusually Sulfated and Oxygenated Steroids from Withania Somnifera,” Phytochemistry, vol. 66, no. 23, pp. 2702–2707, Dec. 2005. doi: 10.1016/j.phytochem.2005.10.001. Abstract
     Four (1, 8–10) and six known (2–7) withanolides were isolated from the leaves of Withania somnifera. Among the new compounds, 10 possessed the rare 3-O-sulfate group with the saturation in A ring and 9 contained unusual 1,4-dien-3-one group. Compound 8 did not have usual 2,3 unsaturation in A ring while 1 had the rare C-16 double bond. The structures of all the compounds were elucidated by spectroscopic methods and chemical transformation.
355. L. Misra, P. Mishra, A. Pandey, R. S. Sangwan, N. S. Sangwan, and R. Tuli, “Withanolides from Withania Somnifera Roots,” Phytochemistry, vol. 69, no. 4, pp. 1000–1004, Feb. 2008. doi: 10.1016/j.phytochem.2007.10.024. Abstract
     Two new and seven known withanolides along with β-sitosterol, stigmasterol, β-sitosterol glucoside, stigmasterol glucoside, α+β glucose were isolated from the roots of Withania somnifera. Among the known compounds, Viscosa lactone B, stigmasterol, stigmasterol glucoside and α+β glucose are being reported from the roots of W. somnifera for the first time. One of the new compounds contained the rare 16β-acetoxy-17(20)-ene the other contained unusual 6α-hydroxy-5,7α-epoxy functional groups in the withasteroid skeleton. The structures were elucidated by spectroscopic methods and chemical transformations.
356. A. Mohanty, J. P. Sahoo, A. U. Acharya, and K. C. Samal, “Ashwagandha (Withania Somnifera) - The Nature’s Gift to Mankind,” Biotica Research Today, vol. 3, no. 2, pp. 122–124, Feb. 2021. https://www.biospub.com/index.php/biorestoday/article/view/756. Abstract
     Ashwagandha is a prominent herb in Indian Ayurvedic medicine and has become a popular supplement due to its health benefits. Limited evidence suggests that Ashwagandha reduces blood sugar levels through its effects on insulin secretion and sensitivity. Its supplements may help lower cortisol levels in chronically stressed individuals. It may help reduce depression. It has been shown to increase muscle mass, reduce body fat, and increase strength in men. It may help reduce the risk of heart disease by decreasing cholesterol and triglyceride levels. It supplements may improve brain function, memory and the ability to perform tasks. Although Ashwagandha is safe for most people, specific individuals should not use it unless authorized by their healthcare provider.
357. A. Molchanova et al., “Antioxidant Properties and Elemental Composition of Withania Somnifera L.,” Agriculture and Food, 2019. https://www.scientific-publications.net/get/1000034/1564599629890995.pdf. Abstract
     Withania somnifera (L.) Dunal is popular in the traditional medicine of many areas of the world due to its high biological activity. Research was carried out in order to characterize the plants of this species in terms of antioxidant content and activity, as well as macro- and microelements concentration in leaves and roots plants grown along Crimea sea coast, which receive an intensive element transfer from the sea surface. The leaves of Withania somnifera showed higher levels of polyphenols, flavonoids and total antioxidant activity compared to roots (7.52 mg eq. GA g-1 d.w., 2.30 mg eq. quercetin g-1 d.w. and 38.2 mg eq. GA g-1 d.w. in leaves; 3.83 mg eq. GA g-1 d.w; 0.48 mg eq. quercetin g-1 d.w. and 10.8 mg eq. GA g-1 d.w. in the roots). Withanolides content in roots reached 0.62 %. The determination of 25 macro- and microelements revealed that Withania somnifera intensively accumulated Ca, K, Mg, P, B, I, Li, Mn, Mo, Si and Zn in leaves, whereas the concentration of V, Cr, Fe and Al in roots exceeded those recorded in leaves by 5.5, 5.86, 4.35 and 5.26 times respectively. The highest difference between leaves and roots concentration was detected for Li whose content in leaves was 10 times higher than in roots. Among other cultivated herbs (14 species belonging to Artemisia and Myrthus), Withania somnifera showed the highest levels of Li which makes it suppose this species is a good Li source for humans. Moreover, the leaves of W. somnifera contain high levels of iodine.
358. C. A. Moore and E. G. Eglington, “The Nature of the Inheritance of Permanently Induced Changes in Nicotiana Rustica,” Heredity, vol. 31, no. 1, pp. 112–118, Aug. 1973. doi: 10.1038/hdy.1973.63. Abstract
     Characters measured on the F5 generation of two crosses, p3 × nil3 and nk2 × nil1 between conditioned lines Nicotiana rustica, were analysed to study (1) the persistence of the segregation between F1 individuals and (2) the conventional segregation in the later generations. Significant F1 segregation was found in four characters in the cross nk2 × nil1 height at flowering time, the length of the branch in the axil of the eighth leaf at flowering time, and the length and width of the eighth leaf, while it was nearly significant for the height of the eighth leaf at flowering time. This item was not significant for any character in the p3 × nil3 cross. A standard biometrical genetical model was found to fit in the majority of cases confirming conventional segregation at the F2, F3 and F4 levels. Principle components analysis extracted three principle components from the seven characters measured in the F5 generation. The three components were the same for each cross and this information together with estimates of the numbers of effective factors segregating in these crosses suggested that for both crosses the parental lines differed by about 13 effective factors.
359. H. N. Murthy and N. Praveen, “Carbon Sources and Medium pH Affects the Growth of Withania Somnifera (L.) Dunal Adventitious Roots and Withanolide A Production,” Natural Product Research, vol. 27, no. 2, pp. 185–189, Jan. 2013. doi: 10.1080/14786419.2012.660691. Abstract
     This work deals with optimisation of adventitious root suspension culture of Withania somnifera (L.) Dunal (Solanaceae) for the production of biomass and withanolide A and we investigated the effect of carbon source and the initial medium pH on growth and production of withanolide A in adventitious root cultures of Withania somnifera. A 2% sucrose concentration was found to be best for both biomass (113.58 g L−1 fresh weight (FW) and 11.33 g L−1 dry weight (DW)) and secondary metabolite accumulation (8.93 mg g−1 DW) in the tested range of concentration (1–8%). The biomass of adventitious roots was optimal when the initial medium pH was 5.8 (113.26 g L−1 FW and 11.33 g L−1 DW) but the withanolide A production was highest at the medium pH level of 5.5 (9.09 mg g−1 DW).
360. H. N. Murthy et al., “Establishment of Withania Somnifera Hairy Root Cultures for the Production of Withanolide A,” Journal of Integrative Plant Biology, vol. 50, no. 8, pp. 975–981, 2008. doi: 10.1111/j.1744-7909.2008.00680.x. Abstract
     Withania sominifera (Indian ginseng) was transformed by Agrobacterium rhizogenes. Explants from seedling roots, stems, hypocotyls, cotyledonary nodal segments, cotyledons and young leaves were inoculated with A. rhizogenes strain R1601. Hairy (transformed) roots were induced from cotyledons and leaf explants. The transgenic status of hairy roots was confirmed by polymerase chain reaction using nptII and rolB specific primers and, subsequently, by Southern analysis for the presence of nptII and rolB genes in the genomes of transformed roots. Four clones of hairy roots were established; these differed in their morphology. The doubling time of faster growing cultures was 8–14 d with a fivefold increase in biomass after 28 d compared with cultured, non-transformed seedling roots. MS-based liquid medium was superior for the growth of transformed roots compared with other culture media evaluated (SH, LS and N6), with MS-based medium supplemented with 40 g/L sucrose being optimal for biomass production. Cultured hairy roots synthesized withanolide A, a steroidal lactone of medicinal and therapeutic value. The concentration of withanolide A in transformed roots (157.4 μg/g dry weight) was 2.7-fold more than in non-transformed cultured roots (57.9 μg/g dry weight).
361. H. N. Murthy and N. Praveen, “Influence of Macro Elements and Nitrogen Source on Adventitious Root Growth and Withanolide-A Production in Withania Somnifera (L.) Dunal,” Natural Product Research, vol. 26, no. 5, pp. 466–473, Mar. 2012. doi: 10.1080/14786419.2010.490914. Abstract
     Withania somnifera (L.) Dunal. (Indian ginseng) is an important medicinal plant which yields pharmaceutically active compounds, namely withanolides. This study deals with the optimisation of the adventitious root suspension culture of W. somnifera for the production of biomass and withanolide-A. We investigated the effects of macro elements (NH4NO3, KNO3, CaCl2, MgSO4 and KH2PO4) and nitrogen source [ ] of Murashige and Skoog (MS) medium on the accumulation of biomass and withanolide-A content. The highest accumulation of fresh and dry biomass (127.52 and 12.45 g L−1) was recorded in the medium with 0.5× concentration of NH4NO3 and the highest production of withanolide-A was recorded in the medium with 2.0× KNO3 (14.00 mg g−1 DW). The adventitious root growth was greater when the concentration was higher than that of and the withanolide-A production was highest in the absence of . Maximum biomass growth was achieved at ratio of 14.38 : 37.60, while withanolide-A production was greatest (11.76 mg g−1 DW) when the ratio was 0.00 : 18.80 mM. The results of this study are useful for scale-up processes.
362. P. D. Naik and M. R. L, “Quantitative Analysis of Secondary Metabolites of Withania Somnifera and Datura Stramonium - Count Search,” International Journal of Science and Research, vol. 4, no. 3, pp. 300–303, Mar. 2015. https://www.ijsr.net/. Abstract
     Medicinal plants are importance resource of drugs of traditional system of medicine. Secondary metabolites are the chemical constituents present in the plants and are important in determining medicinal properties of the plant. The present paper deals with the methanol extract, n - Hexane extract of Withania somnifera, Datura stramonium were screened for the quantitative analysis by standard procedure and subjected to analysis by UV Spectrophotometer. Results shows that Steroids and Alkaloids are in higher amount than the phenols & flavonoids. This indicate that seasonal variation played a key role in increasing secondary metabolites rather than the phenological stage of the plant. - Count Search
363. F. Nakanishi, K. Sasaki, and K. Shimomura, “Kinetics of Littorine Content in Various Developing Stages of Regenerates of Atropa Belladonna,” Plant Cell Reports, vol. 19, no. 10, pp. 1021–1026, Oct. 2000. doi: 10.1007/s002990000231. Abstract
     Aseptically propagated regenerates were cultivated in a hydroponic apparatus, a phytotron or in the field, and their growth and littorine contents were investigated. No littorine was detected in aseptic regenerates cultured on solidified Murashige and Skoog medium, nor was it found in leaves under the three conditions tested. In roots, it was common features to all three conditions tested that littorine increased dramatically after transplantation from culture tubes and was a major alkaloid up to week 4; subsequently the littorine contents varied depending on the cultivation conditions. Roots cultivated in the field showed a marked thickening and rapid disappearance of littorine; those cultivated in the hydroponic apparatus were thin and maintained a high level of littorine for a long time. In a plant cultivated for 16 weeks in a pot, littorine content in the roots decreased with increasing root diameter.
364. A. G. Namdeo et al., “Metabolic Characterization of Withania Somnifera from Different Regions of India Using NMR Spectroscopy,” Planta Medica, vol. 77, no. 17, pp. 1958–1964, Nov. 2011. doi: 10.1055/s-0031-1279997. Abstract
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365. S. Nathiya, D. Pradeepa, T. Devasena, and K. Senthil, “Studies on the Effect of Sucrose, Light and Hormones on Micropropagation and in Vitro Flowering of Withania Somnifera Var. Jawahar-20,” J. Anim. Plant Sci., p. 7. Abstract
     Withania somnifera (solanaceae) popularly, known as “Ashwagandha” is one of the major herbal components of geriatric tonics mentioned in Indian system of medicine. In the present study, effect of various factors on in vitro germination such as sucrose concentration (1%, 2% and 3%) and light intensity (light and dark) in half strength Murashige Skoog medium (1/2 MS) has been studied. 1/2 MS supplemented with 2% sucrose incubated in dark at 25˚C was found to give maximum rate of germination compared to ½ MS supplemented with 1% and 3% sucrose. Different hormonal concentrations were used to test the efficiency of micro shoot formation at different time intervals after inoculation for a period of 90 days with regular subculturing. The maximum number of microshoots was developed in MS media supplemented with 0.5 BAP and 0.05 TDZ followed by 1.0 BAP and 0.05 TDZ. The well-propagated shoots were transferred to elongation media. MS medium supplemented with 1.0 KN and 0.05 BAP showed maximum elongation of shoots. The multiple shoots obtained from 0.5 BAP and 0.05 TDZ; 1 BAP and 0.05 TDZ were transferred to flower induction medium. Different hormonal combination were used to induce flowering, MS medium supplemented with combination of 1 KN and 0.05 BAP exhibits 66% of flowering followed by 0.1 BAP. The matured and immatured pollens in the flowers were observed by pollen staining which contribute to the mode of transfer of genes from one generation to another. Thus the number of matured pollens were more in the in vitro flowers similar to in vivo flowers. In this study an effective methodology for the in vitro seed germination, propagation and flowering was standardized.
366. P. Nayak, S. Upadhyaya, and A. Upadhyaya, “HPTLC Method for Analysis of Withaferin-A in Ashwagandha (Withania Somnifera),” JPC - Journal of Planar Chromatography - Modern TLC, vol. 22, no. 3, pp. 197–200, Jun. 2009. doi: 10.1556/jpc.22.2009.3.7. Abstract
     A densitometric high-performance thin layer chromatographic (HPTLC) method has been established for analysis of withaferin-A in Withania somnifera . The analyte was extracted with methanol. Withaferin-A standard and sample was spotted by use of a sample applicator. The plates were developed with toluene-ethyl acetate-formic acid 5:5:1 as mobile phase. Quantitative evaluation was performed by measuring the absorbance of the analyte zones at 200 nm in reflectance mode. The method was shown to have the selectivity, accuracy, precision, and high sample throughout useful for routine analysis of the preparation in industrial quality control and regulatory laboratories.
367. S. A. Nayak et al., “In Vitro Plant Regeneration from Cotyledonary Nodes of Withania Somnifera (L.) Dunal and Assessment of Clonal Fidelity Using RAPD and ISSR Markers,” Acta Physiologiae Plantarum, vol. 35, no. 1, pp. 195–203, Jan. 2013. doi: 10.1007/s11738-012-1063-2. Abstract
     An efficient large-scale clonal propagation protocol has been described for Withania somnifera (L.) Dunal, a valuable medicinal plant, using cotyledonary nodes derived from axenic seedlings. Murashige and Skoog’s (Physiol Plant 15:473–497, 1962) (MS) medium supplemented with 1.0 mg l−1N6-benzyladenine (BA) was found to be optimum for production of multiple shoots (100 % shoot proliferation frequency and 16.93 shoots per explant). Successive shoot cultures were established by repeatedly sub-culturing the original cotyledonary node on a fresh medium after each harvest of newly formed shoots. Multiple shoot proliferation was also achieved from nodal segments derived from in vitro raised shoots on MS medium augmented with 1.0 mg l−1 BA. Regenerated shoots were best rooted (95.2 %, 38.7 roots per shoot) in half-strength MS medium supplemented with 1.0 mg l−1 indole-3-butyric acid. The plantlets were successfully acclimated and established in soil. Random amplified polymorphic DNA and inter-simple sequence repeats analysis revealed a homogeneous amplification profile for all micropropagated plants analyzed validating the genetic fidelity of the in vitro regenerated plants.
368. A. Nefzi, R. A. B. Abdallah, H. Jabnoun-Khiareddine, N. Ammar, and M. Daami-Remadi, “Ability of Endophytic Fungi Associated with Withania Somnifera L. to Control Fusarium Crown and Root Rot and to Promote Growth in Tomato,” Brazilian Journal of Microbiology, vol. 50, no. 2, pp. 481–494, Apr. 2019. doi: 10.1007/s42770-019-00062-w. Abstract
     Fusarium crown and root rot (FCRR), caused by Fusarium oxysporum f. sp. radicis-lycopersici (FORL), is a soilborne tomato disease of increased importance worldwide. In this study, Withania somnifera was used as a potential source of biological control and growth-promoting agents. Seven fungal isolates naturally associated with W. somnifera were able to colonize tomato seedlings. They were applied as conidial suspensions or a cell-free culture filtrate. All isolates enhanced treated tomato growth parameters by 21.5–90.3% over FORL-free control and by 27.6–93.5% over pathogen-inoculated control. All tested isolates significantly decreased by 28.5–86.4% disease severity over FORL-inoculated control. The highest disease suppression, by 86.4–92.8% over control and by 81.3–88.8% over hymexazol-treated control, was achieved by the I6 isolate. FORL radial growth was suppressed by 58.5–82.3% versus control when dual cultured with tested isolates and by 61.8–83.2% using their cell-free culture filtrates. The most active agent was identified as Fusarium sp. I6 (MG835371), which displayed chitinolytic, proteolytic, and amylase activities. This has been the first report on the potential use of fungi naturally associated with W. somnifera for FCRR suppression and for tomato growth promotion. Further investigations are required in regard to mechanisms of action involved in disease suppression and plant growth promotion.
369. N. Nigam et al., “Exploring the Benefits of Biochar over Other Organic Amendments for Reducing of Metal Toxicity in Withania Somnifera,” Biochar, vol. 1, no. 3, pp. 293–307, Sep. 2019. doi: 10.1007/s42773-019-00023-4. Abstract
     Biochar is considered as a good metal sequester and ameliorates the metal toxicity and uptake in plants. However, its effectiveness over other organic amendments is not well discussed. The aim of this study is to compare the biochar amendments alleviating the levels of Pb and Cd in Withania somnifera L. Dunal with other organic manure. Farmyard manure, vermicompost, and biochar were applied in the field spiked with highly available Pb and Cd. Metal bioavailability and accumulation, plant growth and plant physiology, antioxidant enzymes and metabolite content of W. somnifera along with soil properties were evaluated in all treatments. Study indicates that the application of organic manures significantly alleviated the metal levels (33–72%) in the plant in comparison with control. The herbage yield was significantly higher (33–69%) under different organic manures as compared to the metal treatments. Secondary metabolite contents and antioxidant enzymes were higher in metal treatments in comparison with control. Organic manures were not only able to restrain the Pb and Cd in soil but also enhanced the soil microbial activities. Results indicate that biochar amendments were more promising than farmyard manure and vermicompost due to the presence of more stable carbon in biochar and more alleviation in metal uptake. In addition, net profit in the cultivation of W. somnifera was higher for biochar amendments (50% higher) compared to control. The study recommended that biochar could be a better option for commercial and safer production of W. somnifera.
370. \relax S. D. Ninama, \relax K. M. Gediya, \relax M. K. Rathwa, and \relax G. M. Vaghela, “Effect of Integrated Nutrient Management on Growth and Yield Parameters of Rustica Tobacco (Nicotiana Rustica L.) and Its Residual Impact on Succeeding Summer Green Gram (Vigna Radiata L.),” The Pharma Innovation Journal, vol. 11, no. 9, pp. 2520–2528, 2022. https://www.thepharmajournal.com/archives/?year=2022&vol=11&issue=9&ArticleId=15853. Abstract
     The present research work entitled “Effect of integrated nutrient management on growth and yield parameters of rustica tobacco (Nicotiana rustica L.) and its residual impact of succeeding summer green gram (Vigna radiata L.)” a field experiment was conducted during rabi and summer season of years 2020-21 and 2021-22 at Bidi Tobacco Research Station, Anand Agricultural University, Anand, Gujarat. The experimental field had an even topography with a gentle slope having good drainage and sandy loam in texture. The soil of the experimental field at 0-15 cm depth was low in organic carbon and available nitrogen, medium in available phosphorus and potassium and slightly alkaline in reaction. The ten integrated nutrient management treatments viz., T1: 100% RDF (200-00-00 kg/ha), T2: 75% RDF + 25% N from FYM, T3: 75% RDF + 25% N from poultry manure, T4: 75% RDF + 25% N from castor cake, T5: 50% RDF + 50% N from FYM, T6: 50% RDF + 50% N from poultry manure, T7: 50% RDF + 50% N from castor cake, T8: 50% RDF + 25% N from FYM + Azotobacter, T9: 50% RDF + 25% N from poultry manure + Azotobacter, T10: 50% RDF + 25% N from castor cake + Azotobacter were tested in Randomized Block Design with four replications. Rustica tobacco variety GCT 3 was considered as main rabi crop and green gram variety GAM 5 was considered as summer residual crop. The experiment was conducted on the same site during both the years without changing randomization of treatments. Results of the experiment showed that growth parameters viz. plant height of rustica tobacco at 30, 60 DATP and at harvest was found non-significant due to influence of integrated nutrient management treatments during the years 2020-21, 2021-22 and on pooled basis. However, pooled analysis at 30 DATP showed significant result. Integrated nutrient management manifested their non-significant effect on leaf length and leaf width of rustica tobacco recorded at 30, 60 DATP and at harvest for the years 2020-21, 2021-22 and in pooled analysis under application of 75% RDF + 25% N from poultry manure (T3), respectively. In case of dry weight per unit leaf area the result was found non-significant during both the years (202221 & 2021-22), but pooled analysis showed significant result in rustica tobacco. Cured leaf yield of rustica tobacco manifested significant result during both years (2022-21 & 2021-22) and in pooled analysis under application of 75% RDF + 25% N from poultry manure (T3), respectively. In residual summer green gram plant height did not affected significantly due to integrated nutrient management treatments during years 2021, 2022 and on pooled basis. However, number of branches per plant and length of pods were found non-significant during both individual years (2021 and 2022) in residual summer green gram. Although, in pooled analysis number of branches per plant (5.39) and length of pods (8.21 cm) showed their significant influence in residual summer green gram under application of 50% RDF + 50% N from FYM (T5). Whereas in yield parameters of residual summer green gram viz. number of pods per plant, number of seeds per pod, test weight and harvest index was found non-significant due to integrated nutrient management during both the years (2021 & 2022) and in pooled analysis. However, number of seeds per pod was found significant during pooled analysis. Seed and haulm yield of residual summer green gram was found significant due to various integrated nutrient management during both the individual years (2021 & 2022) and in pooled analysis under application of 50% RDF + 50% N from FYM (T5). Interaction effect (Y x T) was found non-significant on all the growth parameters during the experiment.
371. S. S. Nittala and D. Lavie, “Chemistry and Genetics of Withanolides in Withania Somnifera Hybrids,” Phytochemistry, vol. 20, no. 12, pp. 2741–2748, Jan. 1981. doi: 10.1016/0031-9422(81)85278-8. Abstract
     Hybrid plants of Withania somnifera from cross-pollinations of either chemotypes II or III (Israel) and Indian I (Delhi) have been examined. From both hybrids, 14β-hydroxywithanone (5α,14β,17α-trihydroxy-6α,7α-epoxy-1-oxo-22 R-witha-2,24-dienolide) has been identified. This compound is the first example of a 14β-substitution among withanolides. From the second hybrid three additional new compounds were characterized: a 2,4,6-trien-1-one (14α,2OαF-dihydroxy-i-oxo-22 R-witha-2,4,6,24-tetraenolide), a 14α-hydroxywithanone (5α,14α,17α-trihydroxy-6α,7α-epoxy-1-oxo-22 R-witha-2,24-dienolide) and a 6β,7β-epoxywithanone (5α,14α,17α-trihydroxy-6β,7β-epoxy-1-oxo-22 R-witha-2,24-dienolide). An analysis of the inheritance characteristics of various substituents on the withanolide skeleton was based on the occurrence in per cent of each substituent in relation to the total withanolide content in the hybrid plants and their respective parents.
372. S. S. Nittala, V. V. Velde, F. Frolow, and D. Lavie, “Chlorinated Withanolides from Withania Somnifera and Acnistus Breviflorus,” Phytochemistry, vol. 20, no. 11, pp. 2547–2552, Jan. 1981. doi: 10.1016/0031-9422(81)83091-9. Abstract
     The structure of a new chlorinated withanolide isolated from the hybrid plants of Withania somnifera, chemotypes III (Israel) by Indian I, is established as the chlorohydrin of withanolide D (6α-chloro-4β,5β,20αF-trihydroxy-l-oxo-22R-witha-2,24-dienolide). Two other known chlorinated withanolides had been isolated from different sources and additional data are provided, including the X-ray diffraction study of 4-deoxyphysalolactone (chlorohydrin of withanolide E). All available data are used for comparative analysis of the six known chlorinated withanolides. The origin of the chlorine atom in these compounds in the plants has been determined by carrying out a simple reaction of withanolide D with NaCl on silica gel.
373. A. Niyaz and E. N. Siddiqui, “Seed Germination of Withania Somnifera (L.) Dunal,” European Journal of Medicinal Plants, pp. 920–926, May 2014. doi: 10.9734/EJMP/2014/8916.
374. “Ashwagandha Cultivation, Planting, Care and Harvesting | Agri Farming.” Jan-2016. https://www.agrifarming.in/ashwagandha-cultivation. Abstract
     A step by step guide for growing ashwagandha herb. It covers ashwagandha cultivation practices, economics, project report, planting, care, and harvesting.
375. “Assessment of Genetic Variability and Character Association in Tobacco (Nicotiana Tabacum L.) Genotypes.” Abstract
     The present study was carried out to study yield and yield attributing traits in tobacco, to deliver information regarding the amount of variability, broad-sense heritability, genetic advance over mean and traits association. The experiment was laid out during Rabi 2020 in an Augmented Block Design with 242 genotypes and 4 checks. The box plot analysis revealed that maximum variability for all the traits and the Analysis of variance revealed that the magnitude of the phenotypic coefficient of variance is slightly higher than the genotypic coefficient of variance for all characters studied. Furthermore, a high amount of broad-sense heritability and genetic advance over mean were observed for traits like plant height, number of leaves per plant, internode length, leaf length, leaf width, and yield. This implies that direct selection for these traits might lead to significant progress in discovering superior tobacco genotypes. The character association study revealed that yield was positively and significantly correlated with plant height, the number of leaves per plant, leaf length, leaf width, and internode length hence, selection for these traits will help improve yield.
376. “Cold Tolerance of Tobacco Seedlings,” Tabakpflanzer Osterreichs, vol. 25, no. 69, pp. 5–8, Jan. 1974. https://eurekamag.com/research/000/318/000318268.php. Abstract
     In pot trials, tobacco seedlings were treated with 0-1500 ppm Ethrel before being placed in a growth cabinet and being subjected several days later...
377. “Comparison of Individual Randomisation and Plot Design for Assessing Genetic and Environmental Variation in N,” p. 11, 2018. Abstract
     This study aimed to investigate the effects of individual plant and plot randomizations on the expression of genetic variability for five characters among the generations of a selfing series which have been derived from a cross between V2 and V12 pure breeding varieties of Nicotiana. rustica.
378. “Curing Burley Tobacco from an Automated Harvesting System.” . https://doi.org/10.13031/2013.26216.
379. “Diffusion of Moisture from Burley Tobacco Leaves During Curing.” . https://doi.org/10.13031/2013.33676.
380. “Harvesting and Curing Flue-Cured Tobacco - University of Georgia.” . https://esploro.libs.uga.edu/esploro/outputs/report/Harvesting-and-curing-flue-cured-tobacco/9949316227802959?institution=01GALI_UGA.
381. “How to Grow Mandragora Officinarum | World Seed Supply.” Aug-2014. https://www.worldseedsupply.com/tag/how-to-grow-mandragora-officinarum/.
382. “New Constituents of Atropa Belladonna.” . https://www.proquest.com/openview/58dc0df4a061f713957c95a8972110c9/1?pq-origsite=gscholar&cbl=41445. Abstract
     Explore millions of resources from scholarly journals, books, newspapers, videos and more, on the ProQuest Platform.
383. “Nicotiana Plumbaginifolia and Nicotiana Rustica Var Asiatica Hybrids with Nicotiana Tabacum,” Genetika i Selektsiya, vol. 10, no. 1, pp. 50–57, Jan. 1977. https://eurekamag.com/research/005/983/005983680.php. Abstract
     Hybrids were obtained of the N. plumbaginifolia .times. N. tabacum reciprocal crosses and of the N. rustica var. asiatica .times. N. tabacum cross....
384. https://www.indianjournals.com/ijor.aspx?target=ijor:ija&volume=54&issue=4&article=014.
385. https://www.indianjournals.com/ijor.aspx?target=ijor:ijpgr&volume=32&issue=2&article=045.
386. https://www.indianjournals.com/ijor.aspx?target=ijor:ijh&volume=61&issue=2&article=029.
387. “Seed Germination Database - Perennials - D to N.” . https://tomclothier.hort.net/page03.html.
388. “Study of Withania Somnifera with the Spatial Reference of Phytochemical, FTIR and Flavonoids Quantification.” . https://web.s.ebscohost.com/abstract?direct=true&profile=ehost&scope=site&authtype=crawler&jrnl=09767126&AN=75332302&h=UMAIXtJcPGMdZq5W4ne%2fhpY3QpaJs7ACwLiQ2a9iig8nSZOh5tHM0oh5yF9V1R4q63eco83Ln1R6UIhcHet3fw%3d%3d&crl=c&resultNs=AdminWebAuth&resultLocal=ErrCrlNotAuth&crlhashurl=login.aspx%3fdirect%3dtrue%26profile%3dehost%26scope%3dsite%26authtype%3dcrawler%26jrnl%3d09767126%26AN%3d75332302.
389. “TR_redirect – Defense Technical Information Center.” . https://discover.dtic.mil/tr_redirect/.
390. “Trace Metal Analysis in Withania Somnifera - ProQuest.” . https://www.proquest.com/openview/2cd19dcbb0a24e646bc63784590132b5/1?pq-origsite=gscholar&cbl=2050705. Abstract
     Explore millions of resources from scholarly journals, books, newspapers, videos and more, on the ProQuest Platform.
391. “Tropane Alkaloid Biosynthesis in Atropa Belladonna - ProQuest.” . https://www.proquest.com/openview/25d93dfe480cb638695aed4ca87fa06d/1?pq-origsite=gscholar&cbl=18750&diss=y. Abstract
     Explore millions of resources from scholarly journals, books, newspapers, videos and more, on the ProQuest Platform.
392. “In Vitro Mass Propagation and Green House Establishment of Withania Somnifera (L.) Dunal (Solanaceae) and Eomparison of Growth and Comparison of Growth and Chemical Compounds of Tissue Cultured and Seed Raised Plants,” 2009. http://dr.lib.sjp.ac.lk/handle/123456789/903. Abstract
     Withania somnifera (L.) Dunal (Solanaceae) is normally propagated by seeds. The percentage germination is low, due to the presence of certain inhibitory compounds in the fruit. Among the local and Indian cultivars, the Indian cultivar is preferred by the pharmaceutical industry due to the starchy nature of roots. The local cultivar is listed as a threatened plant. In the present study, W. somnifera was successfully mass propagated in vitro, acclimatized and compared with seed raised plants. The best callus production was observed in Murashige and Skoog (MS) medium supplemented with 1.0 μM kinetin (Kin), 4.5 μM benzyl amino purine (BAP), and 1.5 μM naphthelene acetic acid (NAA) within a 14 day dark period. Shoot initiation was observed in calli produced from shoot tips and nodal segments cultured in the above medium but not from the calli produced from leaf pieces. The highest shoot multiplication was observed in calli from nodal segments cultured in the presence of 9.0 μM BAP and 1.0 μM indole-3-actic acid (IAA) (11.30±1.60). Growth regulator free MS medium was the best medium for rooting. In vitro produced plants were acclimatized successfully (80%) in a potting mixture of river sand: top soil: compost (2: 1: 1). The rate of photosynthesis was higher in tissue cultured plants at three months (4.86±0.40 and 5.67±0.31 for morning and noon respectively) and six months (6.20±0.52 and 6.67±0.33 for morning and noon respectively), while stomatal resistance showed the opposite of that. TLC fingerprints indicated that there was no significant difference in chemical identities (steroids) present in tissue cultured and seed raised plants.
393. “Web Page.” . http://bioweb.uwlax.edu/bio203/s2013/siebold_alic/index.htm.
394. “Withania Somnifera (Ashwagandha, Ashwagandha, Vedic, Indian Ginseng, Indian Winter Cherry, Poison Gooseberry, Winter Cherry) | North Carolina Extension Gardener Plant Toolbox.” . https://plants.ces.ncsu.edu/plants/withania-somnifera/.
395. S. Nyman, “Incorporation of Arginine, Ornithine and Phenylalanine into Tropane Alkaloids in Suspension-Cultured Cells and Aseptic Roots of Intact Plants Oi Atropa Belladonna,” Journal of Experimental Botany, vol. 45, no. 7, pp. 979–986, 1994. doi: 10.1093/jxb/45.7.979.
396. T. Ogino, N. Hiraoka, and M. Tabata, “Selection of High Nicotine-Producing Cell Lines of Tobacco Callus by Single-Cell Cloning,” Phytochemistry, vol. 17, no. 11, pp. 1907–1910, Jan. 1978. doi: 10.1016/S0031-9422(00)88731-2. Abstract
     Single-cell clones of Nicotiana tabacum callus showed wide variation in the production of nicotine. An efficient screening of numerous clones was made possible by the adoption of the ‘cell squash method’ devised for estimating the approximate alkaloid content of a small piece of callus. From these clones, several cell lines with higher yield of nicotine (1.0–3.4% of dry wt) have been isolated by repeated clonal selection.
397. S. Ohta and M. Yatazawa, “Effect of Light on Nicotine Production in Tobacco Tissue Culture,” Agricultural and Biological Chemistry, vol. 42, no. 4, pp. 873–877, Apr. 1978. doi: 10.1080/00021369.1978.10863076. Abstract
     The effect of light on nicotine production in cultured tobacco callus tissues was investigated. Illumination strikingly inhibited the nicotine production even though the growth of the tissues was slightly stimulated by the light. The inhibitory effect of light increased as the intensity and the length of the illumination increased. No decisive difference in nicotine production was observed between the effects of blue light and red light. The depression of nicotine production by light was restored completely when the tissue was transferred to the dark. Therefore, nicotine production is concluded to be regulated by the light just as in the case of plant growth regulators. The inhibitory effect of light was assumed to becaused rather by the inhibition of nicotine biosynthesis than by the promotion of nicotine decomposition.
398. K. Okayasu and M. Notaguchi, “Efficient Establishment of Interfamily Heterograft of Nicotiana Benthamiana and Arabidopsis Thaliana,” in Phloem: Methods and Protocols, J. Liesche, Ed. New York, NY: Springer, 2019, pp. 411–420. doi: 10.1007/978-1-4939-9562-2_31. Abstract
     The grafting technique has been applied to study systemic signaling in plants, especially to investigate whether gene action is graft transmissible and/or gene products such as RNAs and proteins are transported systemically. Here we describe an interfamily heterograft system between Nicotiana benthamiana scion plants and Arabidopsis stock plants for the identification of systemic phloem-mobile signals. Since these plants belong to evolutionary distant families and genome databases are available for both, we can reliably identify mobile substances transported from one to the other plant.
399. N. Orcen and G. R. Nazarİab, “Chromium Tolerance at Seedling Stage of Oriental Tobacco (Nicotiana Tabacum L.) Varieties.,” International Journal of AgriScience, vol. 3, no. 3, pp. 245–252, 2013. https://www.cabdirect.org/cabdirect/abstract/20133131309. Abstract
     The aim of this study was to compare the chromium tolerances of 2 oriental tobacco varieties (Basma and Dubek) during seedling stage. Seeds of each variety were subjected to 0, 10, 100, 150 and 200 µM Cr on MS medium. The effects of the different Cr concentrations and genotypic variation were determined on germination percentage, average number of days for germination, stem and root lengths,...
400. M. Öz, M. S. Fidan, C. Baltaci, O. Ücüncü, and S. M. Karatas, “Determination of Antimicrobial and Antioxidant Activities and Chemical Components of Volatile Oils of Atropa Belladonna L. Growing in Turkey,” Journal of Essential Oil Bearing Plants, vol. 24, no. 5, pp. 1072–1086, Sep. 2021. doi: 10.1080/0972060X.2021.1987334. Abstract
     In this study; the essential oils were obtained from the flowers and leaves of Atropa belladonna L. plants through the hydrodistillation method in a Clevenger type device. The chemical composition of volatile oils was determined by analyzing with the GC-MS/MS instrument. The antioxidant activity of the obtained volatile oils was determined according to total antioxidant content (TAC), free radical scavenging (ABTS) capacity, free radical scavenging (DPPH) amounts, total flavonoid content (TFC), total phenolic content (TPC), and ferric reducing antioxidant power (FRAP) capacity methods. Antimicrobial activities of essential oils were determined through the agar-disc diffusion method. As for the results; volatile oil yields of the A. belladonna were determined as 0.37 % in leaves and 1.57 % in flowers. The highest percentage of chemical compounds in essential oils of the A. belladonna plant were determined as (43.75 %) aldehydes in leaves and as (39.96 %) fatty acids in flowers. In addition, the main constituents found in volatile oils were eicosane (35.92 %) in the leaves and hexadecenoic acid (18.84 %) in the flowers of the A. belladonna. The highest amounts of antioxidant for the test specimens were found as 1.37-1.48 mg QE/100 mL in TFC, as 13.25-21.91 mg AA/100 mL in ABTS, as 51.49-62.87 mg GA/100 mL in TPC, and as 4.71-7.97 mg FeSO4/100 mL in FRAP capacity. As the results of the antimicrobial activities, it was determined that only the leaves of A. belladonna show antimicrobial activity against the mold-yeast and gram-positive and gram-negative bacteria microorganisms among the 17 bacteria and 6 yeast-mold types used in the study.
401. V. P. Oza, P. P. Parmar, D. H. Patel, and R. B. Subramanian, “Cloning, Expression and Characterization of l-Asparaginase from Withania Somnifera L. for Large Scale Production,” 3 Biotech, vol. 1, no. 1, pp. 21–26, Jul. 2011. doi: 10.1007/s13205-011-0003-y. Abstract
     l-Asparaginase (E.C. 3.5.1.1) is used as a therapeutic agent in the treatment of acute childhood lymphoblastic leukemia. It is found in a variety of organisms such as microbes, plants and mammals. In plants, l-asparaginase enzymes are required to catalyze the release of ammonia from asparagine, which is the main nitrogen-relocation molecule in these organisms. An Indian medicinal plant, Withania somnifera was reported as a novel source of l-asparaginase. l-Asparaginase from W. somnifera was cloned and overexpressed in E. coli. The enzymatic properties of the recombinant enzyme were investigated and the kinetic parameters (Km, kcat) for a number of substrates were determined. The kinetic parameters of selected substrates were determined at various pH and the pH- and temperature-dependence profiles were analyzed. WA gene successfully cloned into E. coli BL21 (DE3) showed high asparaginase activity with a specific activity of 17.3 IU/mg protein.
402. V. P. Oza, S. D. Trivedi, P. P. Parmar, and R. B. Subramanian, “Withania Somnifera (Ashwagandha): A Novel Source of L-Asparaginase,” Journal of Integrative Plant Biology, vol. 51, no. 2, pp. 201–206, 2009. doi: 10.1111/j.1744-7909.2008.00779.x. Abstract
     Different parts of plant species belonging to Solanaceae and Fabaceae families were screened for L-asparaginase enzyme (E.C.3.5.1.1.). Among 34 plant species screened for L-asparaginase enzyme, Withania somnifera L. was identified as a potential source of the enzyme on the basis of high specific activity of the enzyme. The enzyme was purified and characterized from W. somnifera, a popular medicinal plant in South East Asia and Southern Europe. Purification was carried out by a combination of protein precipitation with ammonium sulfate as well as Sephadex-gel filtration. The purified enzyme is a homodimer, with a molecular mass of 72 ± 0.5 kDa as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresisand size exclusion chromatography. The enzyme has a pH optimum of 8.5 and an optimum temperature of 37 °C. The Km value for the enzyme is 6.1 × 10−2 mmol/L. This is the first report for L-asparaginase from W. somnifera, a traditionally used Indian medicinal plant.
403. S. Pal et al., “Nitrogen Treatment Enhances Sterols and Withaferin A through Transcriptional Activation of Jasmonate Pathway, WRKY Transcription Factors, and Biosynthesis Genes in Withania Somnifera (L.) Dunal,” Protoplasma, vol. 254, no. 1, pp. 389–399, Jan. 2017. doi: 10.1007/s00709-016-0959-x. Abstract
     The medicinal plant Withania somnifera is researched extensively to increase the quantity of withanolides and specifically withaferin A, which finds implications in many pharmacological activities. Due to insufficient knowledge on biosynthesis and unacceptability of transgenic approach, it is preferred to follow alternative physiological methods to increase the yield of withanolides. Prior use of elicitors like salicylic acid, methyl jasmonate, fungal extracts, and even mechanical wounding have shown to increase the withanolide biosynthesis with limited success; however, the commercial viability and logistics of application are debatable. In this investigation, we tested the simple nitrogeneous fertilizers pertaining to the enhancement of withaferin A biosynthesis. Application of ammonium sulfate improved the sterol contents required for the withanolide biosynthesis and correlated to higher expression of pathway genes like FPPS, SMT1, SMT2, SMO1, SMO2, and ODM. Increased expression of a gene homologous to allene oxide cyclase, crucial in jasmonic acid biosynthetic pathway, suggested the involvement of jasmonate signaling. High levels of WRKY gene transcripts indicated transcriptional regulation of the pathway genes. Increase in transcript level could be correlated with a corresponding increase in the protein levels for WsSMT1 and WsWRKY1. The withaferin A increase was also demonstrated in the potted plants growing in the glasshouse and in the open field. These results implicated simple physiological management of nitrogen fertilizer signal to improve the yield of secondary metabolite through probable involvement of jasmonate signal and WRKY transcription factor for the first time, in W. somnifera besides improving the foliage.
404. J. Panchal and K. M. Gedia, “Effect of Different Management Practices on Growth, Yield Attributes and Yield of Rustica Tobacco.,” Trends in Biosciences, vol. 8, no. 17, pp. 4524–4527, 2015. https://www.researchgate.net/publication/352737292_Effect_of_Different_Management_Practices_on_Growth_Yield_Attributes_and_Yield_of_Rustica_Tobacco. Abstract
     200 kg Ammonium Sulphate ha-1 and manual removal practice were the most effective for securing higher cured leaf yield and yield attributes which resulted into increased cured leaf yield to the tune of 21.11% and 21.35% respectively over control. Among different levels of management through fertilizers, maximum net realization (105533 ha-1) along with BCR value of 4.51 were obtained under...
405. J. Panchal, K. M. Gediya, and J. N. Patel, “Response of Vegetable Intercrops on the Performance of Chewing Tobacco (Nicotiana Rustica L.) Based Intercropping System,” Agricultural Research Journal, vol. 60, no. 2, pp. 177–181, 2023. doi: 10.5958/2395-146X.2023.00028.5.
406. S. S. Pandey et al., “Endophytes of Withania Somnifera Modulate in Planta Content and the Site of Withanolide Biosynthesis,” Scientific Reports, vol. 8, no. 1, p. 5450, Apr. 2018. doi: 10.1038/s41598-018-23716-5. Abstract
     Tissue specific biosynthesis of secondary metabolites is a distinguished feature of medicinal plants. Withania somnifera, source of pharmaceutically important withanolides biosynthesizes withaferin-A in leaves and withanolide-A in roots. To increase the in planta withanolides production, a sustainable approach needs to be explored. Here, we isolated endophytes from different parts of W. somnifera plants and their promising role in in planta withanolide biosynthesis was established in both in-vivo grown as well in in-vitro raised composite W. somnifera plants. Overall, the fungal endophytes improved photosynthesis, plant growth and biomass, and the root-associated bacterial endophytes enhanced the withanolide content in both in-vivo and in-vitro grown plants by modulating the expression of withanolide biosynthesis genes in leaves and roots. Surprisingly, a few indole-3-acetic acid (IAA)-producing and nitrogen-fixing root-associated endophytes could induce the biosynthesis of withaferin-A in roots by inducing in planta IAA-production and upregulating the expression of withanolide biosynthesis genes especially MEP-pathway genes (DXS and DXR) in roots as well. Results indicate the role of endophytes in modulating the synthesis and site of withanolides production and the selected endophytes can be used for enhancing the in planta withanolide production and enriching roots with pharmaceutically important withaferin-A which is generally absent in roots.
407. R. Pandey, A. K. Mishra, S. Tiwari, and A. Kalra, “Nematode Inhibiting Organic Materials and a Strain of Trichoderma Harzianum Effectively Manages Meloidogyne Incognita in Withania Somnifera Fields,” Biocontrol Science and Technology, vol. 21, no. 12, pp. 1495–1499, Dec. 2011. doi: 10.1080/09583157.2011.625396. Abstract
     The usefulness of Trichoderma harzianum was tested along with farmyard manure, cow urine, neem oil seed cake, and vermicompost separately and in combination to manage Meloidogyne incognita in Withania somnifera. A treatment combination of nematode inhibitory vermicompost and T. harzianum was found to be most effective against M. incognita.
408. R. Pandey and A. Kalra, “Root-Knot Disease of Ashwagandha Withania Somnifera and Its Eco-Friendly Cost Effective Management,” Journal of Mycology And Plant Pathology (India), 2003. https://agris.fao.org/search/en/records/64723fde53aa8c8963034954. Abstract
     Ashwagandha (Withania somnifera L.) is an important medicinal plant and a major source of alkaloid and steroidal lactones (withanolide), which are regularly used in pharmaceutical industries. Plant growth retardation and gall formation in the root system indicated the presence of root-knot nematode, which was confirmed as Meloidogyne incognita race-2. Green house trials were conducted to determine the influence of different inoculum levels of Meloidogyne incognita (Kofoid and White) Chitwood on growth and yield of W. somnifera. Various organic materials viz., Neem compound, Mentha distillate, Murraya koengii distillate, Artemisia annua marc and Vermicompost, bio-agents viz., Glomus aggregatum and Trichoderma harzianum were tested individually as well as in different combinations for the management of root-knot nematode, M. incognita on W. somnifera. The experiment results indicated that most of the bio-agents and organic materials alone as well as in combination were root-knot nematode suppressive and enhanced the growth and yield of W. somnifera. Maximum root-knot suppression was noticed in vermicompost and T. harzianum combination followed by Mentha distillate and G. aggregatum. Maximum increase in plant yield was noticed when the soil was amended with Mentha and Murraya koengii distilled waste along with bio-agents.
409. V. Pandey, W. A. Ansari, P. Misra, and N. Atri, “Withania Somnifera: Advances and Implementation of Molecular and Tissue Culture Techniques to Enhance Its Application,” Frontiers in Plant Science, vol. 8, 2017. doi: 10.3389/fpls.2017.01390. Abstract
     Withania somnifera, commonly known as Ashwagandha an important medicinal plant largely used in Ayurvedic and indigenous medicine for over 3,000 years. Being a medicinal plant, dried powder, crude extract as well as purified metabolies of the plant has shown promising therapeutic properties. Withanolides are the principal metabolites, responsible for the medicinal properties of the plant. Availability and amount of particular withanolides differ with tissue type and chemotype and its importance leads to identification characterization of several genes/ enzymes related to withanolide biosynthetic pathway. The modulation in withanolides can be achieved by controlling the environmental conditions like, different tissue culture techniques, altered media compositions, use of elicitors, etc. Among all the in vitro techniques, hairy root culture proved its importance at industrial scale, which also gets benefits due to more accumulation (amount and number) of withanolides in roots tissues of W. somnifera. Use of media compostion and elicitors further enhances the amount of withanolides in hairy roots. Another important modern day technique used for accumulation of desired secondary metabolites is modulating the gene expression by altering environmental conditions (use of different media composition, elicitors, etc.) or through genetic enginnering. Knowing the significance of the gene and the key enzymatic step of the pathway, modulation in withanolide contents can be achieved upto required amount in therapeutic industry. To accomplish maximum productivity through genetic enginnering different means of Withania transformation methods have been developed to obtain maximum transformation efficiency. These standardized transformation procedues have been used to overexpress/silence desired gene in W. somnifera to understand the outcome and succeed with enhanced metabolic production for the ultimate benefit of human race.
410. D. Pandya, A. Mankad, and H. Pandya, “A Review on Withania Somnifera (l.) Dunal- as an Important Ayurveda Plant,” International Association of Biologicals and Computational Digest, vol. 1, no. 1, pp. 138–144, May 2022. doi: 10.56588/iabcd.v1i1.28. Abstract
     Withania somnifera (L.) Dunal is a well-known and important medicinal plant widely used in several indigenous systems of medicine for the treatment of various ailments, viz. asthma, bronchitis, inflammatory diseases, ulcer and stomach problems. Steroidal lactones have been reported as the major phytoconstituents of this species. Different pharmacological experiments in a number of in vitro and in vivo models have convincingly demonstrated the ability of W. somnifera to exhibit anti-inflammatory, anti-oxidative, antimicrobial, anti-anxiety, aphrodisiac, immunomodulation, anti- diabetic, anti-ulcer, anticancer, central nervous system depressant and hepatoprotective activities, lending support to the rationale behind several of its traditional uses. The species is also used to treat some neurological disorders like Parkinson’s and Alzheimer’s. The phytochemicals such as withaferin A, withanolide A and withanolide D isolated from this plant are potential bioactive molecules. Due to the remarkable biological activity of W. somnifera and its constituents, it will be appropriate to develop them as a medicine and make them more potent by chemical modifications and biotransformation. This review has covered botany, chemistry and pharmacology of the plant besides its traditional and folkloric uses.
411. G. Pant and P. Agrawal, “Isolation and Characterization of Indole Acetic Acid Producing Plant Growth Promoting Rhizobacteria from Rhizospheric Soil of Withania Somnifera,” Journal of Biological & Scientific Opinion, vol. 2, pp. 377–383, Dec. 2014. doi: 10.7897/2321-6328.02687.
412. S. Parajuli, D. A. Patel, M. G. Makwana, and D. J. Parmar, “Genetic Diversity Studies on Cured Leaf Yield and Its Components in Rustica Tobacco [Nicotiana Rustica (L.)],” p. 5, 2015. Abstract
     Study on genetic diversity was carried out with 40 genotypes of rustica tobacco (Nicotiana rustica L.) using Mahalanobis D² statistics at Bidi Tobacco Research Station, Anand Agricultural University, Anand (Gujarat) during rabi, 2012-2013. The 40 genotypes were grouped into 16 clusters based on D² analysis. The cluster II and V had maximum and clusters VIII, X, XI, XIII, XIV, XV and XVI had the minimum number of genotypes. The highest inter cluster distance (D2 =1979.3) was observed between cluster XII and XV and the lowest (D2 =133.37) between clusters IIIand VII. Cluster XII (D2 =173.45) had exhibited highest intra cluster distance and the lowest was observed in cluster XIII, X, XI, XIII, XIV, XV and XVI (D2 =0). The character nicotine content (17.46 %), days to flowering (14.73 %), reducing sugar content (11.43%), cured leaf yield (10.90 %) and plant height (10.88 %) contributed much to the total genetic divergence. On the basis of cluster mean, cluster XIII was superior for cured leaf yield (144.76 g) and nicotine content (4.50 %). The maximum number of leaves per plant (16.66) was observed in cluster XIV, while cluster IX sowed superiority for plant height (72.25 cm). The cluster XVI and VII showed highest quality for leaf length (44.46 cm) and leaf breadth (38.53 cm), respectively. The cluster XV had desirable rating in respect to days to flowering (29.33) and days to maturity (105.00). The cluster VIII was best for leaf thickness, while cluster IV and X were found outstanding for chloride content (2.49 %) and reducing sugar content (3.76%), respectively. Thus, the genotypes involved in these clusters may be taken into consideration for better parents for generating variability for the respective character and their rational improvement.
413. P. Parihar and M. Bora, “Effect of Mycorrhiza (Glomus Mosseae) on Morphological and Biochemical Properties of Ashwagandha (Withania Somnifera) (L.) Dunal,” Journal of Applied and Natural Science, vol. 10, no. 4, pp. 1115–1123, Dec. 2018. doi: 10.31018/jans.v10i4.1797. Abstract
     Mycorrhizal inoculation in the plant causing increase in growth and production of phytochemicals is well reported, however little information is available related to the effect of mycorrhiza on morphological and biochemical properties of the medicinal plants like Ashwagandha. The present study is an attempt on diversity analysis in Withania somnifera with an aim to ascertain the nature and extent of genetic diversity present among different accessions in presence of mycorrhiza. The major biochemical constituents of Ashwaganda roots are with nolides which are well known for its medicinal properties. Mycorrhizal associations confer benefits like better nutrition acquisition, enhanced growth, defense enhancement and improved abiotic and biotic stress tolerance in plants. The present investigation was undertaken to assess genetic diversity among five different accessions of W. somnifera using morphological and biochemical markers and the effect of mycorrrhizal inoculation on these marker. The present study concluded that presence of mycorrhiza was effective on plant growth and phytochemical constituents more than non-treated plants. Amongst five selected germplasms IC 283662, JA 134, RAS 23, MPAS 6 and MWS 205 of W. somnifera, JA 134 showed best response in pretext of the selected morphological and biochemical features in presence of mycorrhiza.
414. W. M. Park, G. P. Lee, K. H. Ryu, and K. W. Park, “Transmission of Tobacco Mosaic Virus in Recirculating Hydroponic System,” Scientia Horticulturae, vol. 79, no. 3, pp. 217–226, Feb. 1999. doi: 10.1016/S0304-4238(98)00215-5. Abstract
     The possible pathway of transmission of tobacco mosaic virus (TMV) from diseased plants to healthy plants in two hydroponic systems, deep flow culture (DFC) and container culture (CC), was investigated in this study. The tested plants were tobacco (Nicotiana tabacum cv. Samsun-nn and cv. Xanthi), tomato (Lycopersicon esculentum cv. Alchan), and hot pepper (Capsicum annuum cv. Dahong). The longevity in vitro and the dilution end point of the virus in the hydroponic solution were 5 days and 10−4 (w/v), respectively. When inoculum source plants and tested healthy plants were grown side by side in the same gully or containers, typical TMV symptoms appeared on the tested plants in 2 weeks. The presence and concentration of the virus in the tissues of the tested plants could be detected by DAS-ELISA. It was noticed that roots of the inoculum source plants grafted to those of the tested plants during growing period. However, when both inoculated plants and healthy plants were grown separately, and their root tips were either removed or remained intact, no transmission of the virus from the inoculum source plants to the tested healthy plants occurred even though they shared the same recycled nutrient solution. These results indicate that the virus in a recirculating hydroponic system could be transmitted from virus-diseased plants to healthy plants mainly via root-tip grafting but was hardly transmitted through regularly renewed hydroponic nutrient solution.
415. S. C. Parmar, S. V. Rathod, S. M. Gondaliya, and N. N. Chaudhary, “Influences of Varying Proportion of Urea and Ammonium Sulphate on Yield and Chemical Composition of Rustica Tobacco (Nicotiana Rustica L.) as Well as Chemical Properties of Soil,” The Bioscan, vol. 11, no. 4, pp. 2553–2557, 2016. http://thebioscan.com/supplements/56_7239_S.C.%20PARMAR_Agro.pdf. Abstract
     The present investigation was carried out to find out ideal proportion of urea and Amonium sulphate for the application of 200 kg N ha-1 and thereby different levels of S on yield, quality and chemical composition of rustica tobacco variety GC-1 and related soil properties in middle Gujarat condition. There are six treatments where comprised with four replications applied as urea and AS as follow: T1 100:80, T2 80:20, T3 60:40, T4 40:60, T5 20:80 and T6 00:100. The thickness of the leaf (30.75 mg/cm2) was significantly higher due to T2.. The yield (3203 kg ha-1) was significantly higher with T2 treatment which was at par with T1 while the quality parameters found non-significant. The N, K and S contents were increased up to 3.73, 0.80 and 0.68 % respectively in T5 while P content increased about 0.65 % in T1 treatments. The uptakes of all these macronutrients were significantly higher with T2 and found at par with T1 and T3. The Mn, Zn and Cu content of the leaf was about 232.0, 53.0 and 35.0 ppm respectively which was significantly higher with T6. The available P2O5 and S contents of the soil were significantly influenced by different treatments.
416. J. B. Patel, S. K. Lahiri, and M. B. Shah, “Development of a New Method for Identification and Estimation of Withania Somnifera Root, and a Method for Quantitative Analysis of Withaferin A in Young and Old Roots,” JPC – Journal of Planar Chromatography – Modern TLC, vol. 22, no. 4, pp. 283–286, Aug. 2009. doi: 10.1556/JPC.22.2009.4.8. Abstract
     Withania somnifera, commonly known as Ashwagandha, is a valued herb in Ayurvedic medicine. Root leaves, and preparations of the plant are traditionally used as tonic, hypnotic, sedative and diuretic. W. somnifera mainly contains withanolides which are specific to the Solanaceae family. The biological activity of withanolides, espeeially withaferin A, has been studied extensively. Methods for analysis of withaferin A and withnolide D require acetylation before analysis, or separation time is extremely long. The main problem in identification of withaferin A with other separated withanolides is that all withanolides absorb UV light, so without comparison with a standard it is not possible to identify withaferin A. The objectives of this paper are to present a new method of identification of withaferin A, using a destructive reagent, and an HPTLC method for quantification of the compound. Chromatography on silica with toluene-ethyl acetate-acetone 2:3:3 as mobile phase enabled good resolution of withaferin A without interference from other compounds present in W. somnifera. After spraying with anisaldehyde-sulfuric acid reagent and heating for 15 min at 105°C, characteristic orange fluorescence was observed for withaferin A only among all the spots resolved. When scanned at 214 nm the RF of withaferin A was 0.62. Interestingly, old root did not contain withaferin A whereas young root contained a large amount. The method was validated for accuracy, precision, specificky, linearity, and limits of detection and quantification. The method is simple, sensitive, and precise, and can be used for routine qualitycontrol testing of W. somnifera.
417. H. N. Patel, “Heterosis and Inbreeding Depression in Rustica Tobacco (Nicotiana Rustica L.) for Yield and Quality Parameters over Environments.,” Trends in Biosciences, vol. 10, no. 15, pp. 2742–2744, 2017. https://www.cabdirect.org/cabdirect/abstract/20193314920. Abstract
     The present investigation in rustica tobacco (Nicotiana rustica L.) was carried for the study of magnitude of heterosis and inbreeding depression of four hybrids (F1s) and their F2s developed by crossing of five lines in six generation mean analysis under two artificially created environments viz., low fertility and high fertility levels. The data were recorded on quantitative and qualitative...
418. N. Patel, P. Patel, U. Kumari, S. V. Kendurkar, and B. M. Khan, “Microprojectile Bombardment Assisted Agroinfection Increases Transformation Efficiency of Withania Somnifera (L.),” Research in Biotechnology, p. 12, 2014. Abstract
     Withania somnifera (L.) Dunal, an evergreen shrub distributed throughout the drier parts of India, is well known for its medicinal properties. Three different approaches were employed towards development of an efficient transformation system in Withania somnifera, namely Agrobacterium tumefaciens-mediated transformation, microprojectile bombardment and microprojectile bombardment assisted agroinfection. A. tumefaciens strain GV2260 containing a binary vector pCAMBIA 1301 was used carrying β-Glucuronidase (gus) reporter gene and hygromycin phosphotransferase (hptII) selection gene under the transcriptional control of Cauliflower Mosaic Virus 35S promoter. The apical and nodal segments of about one cm length excised from in vitro grown shoots were chosen as explants for transformation studies. The selection of transgenic plants was attained in the presence of 10 mg L-1 hygromycin B. Analysis of transgenic plants was done by histochemical Gus assay followed by polymerase chain reaction for the presence of gus and hptII genes. The overall transformation efficiencies obtained were 3.86%, 3.62% and 8.71% for A. tumefaciensmediated, microprojectile bombardment and a combination of both the techniques, respectively.
419. H. N. Pateli, “Effect of Biparental Mating and Extent of Genetic Variability in Tobacco (Nicotiana Rustica L.),” Green Farming, vol. 1, no. 2, pp. 158–160, Mar. 2010. Abstract
     S . Proginies of North Carolina design lll from two crosses of to bacco viz.,GC 1 x Rangpur (cross l) and GCT 3 x GC 1 (cross ll)along with theirselfed progenies ( F.) were evaluated in field. Mean characteristics values of progenies of NCD lllwere significantly superior for most of the characters as compared to their F. progenies. The additive variances for yield and plant height in both crosses and for days toflower in cross I and dominance variance for plani height in cross llwere found significant.
420. P. K. Pati, M. Sharma, R. K. Salar, A. Sharma, A. P. Gupta, and B. Singh, “Studies on Leaf Spot Disease of Withania Somnifera and Its Impact on Secondary Metabolites,” Indian Journal of Microbiology, vol. 48, no. 4, pp. 432–437, Dec. 2008. doi: 10.1007/s12088-008-0053-y. Abstract
     During an investigation of the disease profile of Withania somnifera, it was observed that leaf spot is the most prevalent disease. Repeated isolations from infected leaf tissues and pathogenicity tests showed the association of fungal pathogen identified as Alternaria alternata (Fr.) Keissler. Scanning electron microscopy showed various histological changes in the leaf tissues of infected plants. A decrease in total content of reducing sugars (20%) and chlorophyll (26.5%) was observed in diseased leaves whereas an increase was noticed in proline (25%), free amino acids (3%) and proteins (74.3%). High performance thin layer chromatography (HPTLC) analysis of secondary metabolites viz. withanolides, withaferin-A and total alkaloids of the diseased leaves vis-à-vis control revealed reduction in withaferin-A and withanolides contents by 15.4% and 76.3% respectively, in contrast to an increase in total alkaloids by 49.3%, information hitherto unreported in W. somnifera.
421. H. Patidar, V. S. Kandalkar, and S. Nigam, “Estimation of Leaf Area in Asgandh (Withania Somnifera).,” Indian Journal of Agricultural Sciences, vol. 60, no. 4, pp. 263–264, 1990. https://www.cabdirect.org/cabdirect/abstract/19911620194. Abstract
     On the basis of a study in which all the leaves were sampled from 20 plants of each of 5 diverse genotypes, a simple, accurate non-destructive method for estimating total plant leaf area is presented. For each leaf, greatest leaf length is multiplied by greatest leaf width and the result multiplied by the constant 0.7028.
422. S. Paul et al., “Withania Somnifera (L.) Dunal (Ashwagandha): A Comprehensive Review on Ethnopharmacology, Pharmacotherapeutics, Biomedicinal and Toxicological Aspects,” Biomedicine & Pharmacotherapy, vol. 143, p. 112175, Nov. 2021. doi: 10.1016/j.biopha.2021.112175. Abstract
     Withania somnifera (L.) Dunal (Solanaceae) has been used as a traditional Rasayana herb for a long time. Traditional uses of this plant indicate its ameliorative properties against a plethora of human medical conditions, viz. hypertension, stress, diabetes, asthma, cancer etc. This review presents a comprehensive summary of the geographical distribution, traditional use, phytochemistry, and pharmacological activities of W. somnifera and its active constituents. In addition, it presents a detailed account of its presence as an active constituent in many commercial preparations with curative properties and health benefits. Clinical studies and toxicological considerations of its extracts and constituents are also elucidated. Comparative analysis of relevant in-vitro, in-vivo, and clinical investigations indicated potent bioactivity of W. somnifera extracts and phytochemicals as anti-cancer, anti-inflammatory, apoptotic, immunomodulatory, antimicrobial, anti-diabetic, hepatoprotective, hypoglycaemic, hypolipidemic, cardio-protective and spermatogenic agents. W. somnifera was found to be especially active against many neurological and psychological conditions like Parkinson’s disease, Alzheimer’s disease, Huntington’s disease, ischemic stroke, sleep deprivation, amyotrophic lateral sclerosis, attention deficit hyperactivity disorder, bipolar disorder, anxiety, depression, schizophrenia and obsessive-compulsive disorder. The probable mechanism of action that imparts the pharmacological potential has also been explored. However, in-depth studies are needed on the clinical use of W. somnifera against human diseases. Besides, detailed toxicological analysis is also to be performed for its safe and efficacious use in preclinical and clinical studies and as a health-promoting herb.
423. G. J. PAXMAN, “Differentiation and Stability in the Development of Nicotiana Rustica,” Annals of Botany, vol. 20, no. 2, pp. 331–347, Apr. 1956. doi: 10.1093/oxfordjournals.aob.a083526. Abstract
     Stability, or homeostasis, is that property of the organism buffering it against small random fluctuations of the environment and accidents of development. A means of measuring, within the individual, the stability of expression of one foliar and two floral characters is given.Data are presented from a set of diallel crosses among five varieties of Nicotiana rustica, derived from different populations and grown in two successive seasons. This allows stability to be examined over a number of genotypes in the parent and F1 generations, in two differing environments.The leaf characters show some change from node to node, i.e. manifesting a gradual differentiation, as well as random fluctuations which are taken as evidence of instability. Like stability, this differentiation rate varies from genotype to genotype and shows genetic control.Some attempt is made to find to what extent the different phenomena are related.
424. R. C. Pearce, Y. Li, and L. P. Bush, “Calcium and Bicarbonate Effects on the Growth and Nutrient Uptake of Burley Tobacco Seedlings: Hydroponic Culture,” Journal of Plant Nutrition, vol. 22, no. 7, pp. 1069–1078, Jul. 1999. doi: 10.1080/01904169909365696. Abstract
     In soilless production systems, water quality can have a major impact on the growth of plants. It has become evident that moderately alkaline water is a problem for tobacco transplant growers in some regions of Kentucky. To determine the level of bicarbonate (HCO3 −) alkalinity, which is detrimental to burley tobacco transplants, and to better understand the effect of calcium (Ca) and the interaction of HCO3 − and Ca on the growth of burley tobacco transplants grown in a float system, three levels of CaCl2 (25,75, and 125 mg L−1 Ca++) in factorial combination with five levels of HCO3 − (0, 122, 244, 366, and 488 mg HCO3 −L−1) were tested in nutrient solution culture. Four‐week‐old burley tobacco (Nicotiana tabacum L. var. KY‐907) seedlings were transplanted to 18‐L containers filled with aerated Hoagland’s solution with the different levels of calcium and HCO3 − for two weeks. High HCC3 − alkalinity caused root system damage and plant growth inhibition, but did not induce iron (Fe) chlorosis. A significantly lower concentration of Zinc (Zn) was measured in the shoots as HCO3 − levels in solution increased. In the presence of high calcium, plant growth was not significantly improved. No significant interaction of HCO3 − and Ca on growth or nutrient uptake was observed in this study.
425. J. L. Pérez-Rodríguez, R. C. Rodríguez Escriba, G. Y. Lorente González, J. L. González Olmedo, and M. E. Martínez-Montero, “Effect of Desiccation on Physiological and Biochemical Indicators Associated with the Germination and Vigor of Cryopreserved Seeds of Nicotiana Tabacum L. Cv. Sancti Spíritus 96,” In Vitro Cellular & Developmental Biology - Plant, vol. 53, no. 4, pp. 440–448, Aug. 2017. doi: 10.1007/s11627-017-9857-y. Abstract
     Cryopreservation is a valuable technique for the long-term conservation of plant germplasm and complementary to traditional seed storage methods. However, critical factors such as seed moisture content should be optimized before using this technique as a safe strategy for storing seeds such as those of Nicotiana spp. This study aimed to determine the effect of desiccation on physiological and biochemical indicators associated with germination and vigor in cryopreserved seeds of Nicotiana tabacum cv. Sancti Spíritus 96 (SS-96). The germination and vigor of seeds with a range of moisture content were assessed using electrolyte leakage and accelerated aging tests. In addition, these physiological indicators were related to the oxidative state of the seeds, in terms of the rate of O2·− generation and the H2O2 content, and the activity of enzymatic antioxidants superoxide dismutase and catalase. The cryopreserved seeds of N. tabacum SS-96 with a moisture content of 2.1% exhibited higher vigor probably due to the retention of membrane integrity, reflected by lower levels of lipid peroxidation and electrolyte leakage associated with the absence of oxidative stress. The results suggest 2.1% as the optimal moisture content for the storage of seeds of this cultivar, both at cryogenic temperatures and at 5°C.
426. J. D. Phillipson and S. S. Handa, “Hyoscyamine N-Oxide in Atropa Belladonna,” Phytochemistry, vol. 15, no. 5, pp. 605–608, Jan. 1976. doi: 10.1016/S0031-9422(00)94402-9. Abstract
     Separated organs of Atropa belladonna have been examined for their total alkaloid, hyoscyamine and hyoscyamine N-oxide contents during ontogenesis. Marked fluctuations in N-oxide content were observed, the highest being found in the ripe fruit. [G-3H]-atropine was fed to A. belladonna fruits and radioactively labelled hyoscyamine N-oxide isolated.
427. • PK, A. Kumar, A. Ahuja, and M. Kaul, “Effect of Biofertilizers on the Morphology, Root Yield and Biochemical Parameters of Withania Somnifera (L.) Dunal; Journal of Plant Biology, 33 (1&2), Pp. 163-167,” Journal of Plant Biology, vol. 33, pp. 163–167, Jan. 2006.
428. H. B. D. and P.Manivel, “Inheritance of Growth Habit and Berry Colour in Ashwagandha (Withania Somnifera (L.) Dunal) – A Medicinal Plant,” Electronic Journal of Plant Breeding, vol. 5, no. 2, pp. 244–247, Nov. 2014. https://www.ejplantbreeding.org/index.php/EJPB/article/view/126. Abstract
     Genetics studies were studied for the two qualitative character ivz., growth habit (Erect × Procumbent) and for berry (fruit) colour (Yellow × Red) in ashwagandha. The inheritance of the growth habit was controlled by single gene and the procumbent was dominant over the erect type. The gene symbol P for procumbent and p for erect is proposed. Inheritance studies of yellow versus red berry colour indicated that the trait was in control of classical duplicate recessive epitasis and followed the ratio of 9:7 (yellow: red). The berry colour in this cross is controlled by two genes (Y1 and Y2) with complementary recessive epitasis and gene symbols are proposed.
429. H. S. Pooni, R. Foster, and B. Zhao, “Impact of Pollination Time, Seed Size, Position and Maturity on Quantitative Variation in Nicotiana Rustica,” The Journal of Agricultural Science, vol. 128, no. 2, pp. 181–188, Mar. 1997. doi: 10.1017/S0021859696004042. Abstract
     Many biological experiments and almost all quantitative genetic studies draw conclusions from differences between families. Such differences are caused as much by heritable as by non-heritable sources. This paper investigates the impact of seed quality on the expression of quantitative variability among 20 highly inbred lines of Nicotiana rustica using ten seed categories representing different pollination times, seed sizes, seed positions in the capsule and seed maturity. Analysis of variance revealed that while the mean performance of the inbred lines remained more or less the same across all the seed categories, their individual performances differed widely, showing a highly significant interaction between lines and seed categories. This interaction accounted for up to 12% of the total variation and was largely attributable to late pollinations and immature seed. Seed categories were also the main source of heterogeneity among the within-line variances, but only immature seed lowered the correlation between the inbred means. In general, seed quality affected the developmental characters much less than the traits measured at or after flowering, and the expression of genetic variation was not critically affected by seed size or position. Immature seed, on the other hand, generated the highest level of unreliability in the comparative performance of lines and such seeds should therefore not be used in quantitative genetic experiments under any circumstances. Further implications of seed quality effects on the interpretation of genetic components are discussed in detail.
430. H. S. Pooni and J. L. Jinks, “Retrospective Selection and Sources of Superior Inbreds amongst Pedigree Inbred Families of Nicotiana Rustica,” Heredity, vol. 54, no. 2, pp. 255–260, Apr. 1985. doi: 10.1038/hdy.1985.34. Abstract
     A single cycle of selection in the F2, F3, F4 and F7 generations using the overall means of the individual hierarchies as their predicted breeding values, has been applied retrospectively to seven metrical traits scored on 784 F7 inbreds extracted from the cross of varieties 2 and 12 of Nicotiana rustica by the method of pedigree inbreeding. The hierarchical structure within 98 F2 groups each of 8 F7 families is further used to determine the distribution and source of the superior inbreds within each hierarchy. While the efficiency of selection improved steadily the more advanced the generations in which it was applied, superior recombinants originated more frequently in the early than in the late generations. The hierarchical method of inbreeding, however, favours the recovery of recombinants from the advanced generations and in the present case the two balance each other exactly. The improvement in response to selection in the advanced generations is completely associated with the relative reductions in error variances and is, therefore, not attributable to any genuine change in the genetic covariances.
431. G. C. Popescu and M. Popescu, “Effects of Different Potting Growing Media for Petunia Grandiflora and Nicotiana Alata Link & Otto on Photosynthetic Capacity, Leaf Area, and Flowering Potential,” Chilean journal of agricultural research, vol. 75, no. 1, pp. 21–26, Mar. 2015. doi: 10.4067/S0718-58392015000100003.
432. V. T. Popova et al., “Chemical Constituents in Leaves and Aroma Products of Nicotiana Rustica L. Tobacco,” International Journal of Food Studies, vol. 9, no. 1, Apr. 2020. doi: 10.7455/ijfs/9.1.2020.a2. Abstract
     Nicotiana rustica L. (Aztec tobacco) is the only Nicotiana species, except common tobacco (N. tabacum L.), which is cultivated for tabacco products. The leaves of N. rustica, however, accumulate various specialized metabolites of potential interest. Therefore, the objective of this study was to evaluate certain classes of metabolites (by HPLC and GC-MS) in the leaves, the essential oil (EO), concrete and resinoid of N. rustica. Three pentacyclic triterpenes were identified in the leaves (by HPLC): betulin (252.78 µg g-1), betulinic (182.53 µg g-1) and oleanolic  (69.44 µg g-1) acids. The dominant free phenolic acids in the leaves (by HPLC) were rosmarinic (4257.38 µg g-1) and chlorogenic (1714.40 µg g-1), and conjugated forms of vanillic (3445.71 µg g-1), sinapic (1963.11 µg g-1), and syringic (1784.96 µg g-1). The major flavonoids in the leaves were luteolin (960.44 µg g-1), apigenin (880.66 µg g-1) and hyperosid (780.72 µg g-1). The GS-MS profiling of the EO identified 19 components and the major ones were phytol (43.68 %), solanone (5.54 %), cis-5-butyl-4-methyldihydrofuran-2(3H)-one (5.23 %), dihydro-β-ionone (4.25 %), α-ionene (3.54 %),and β-damascenone (3.03 %). The major volatiles in the concrete were  isoamyl alcohol (28.82 %), oxynicotine (9.02 %), phytol (7.80 %), 4-mеthyl-1-penthanol (6.33 %), cotinine (5.55 %) and 3-metyl-3-penthanol (4.09 %). Resinoid composition was dominant by nicotine (39.75 %), phytol (11.23 %), eicosane (4.88 %), diethyl phthalate (4.19 %), dibutyl phthalate (3.48 %) and solanone (3.27 %). Concrete and resinoid showed weak antibacterial activity . These results create grounds for considering N. rustica as a source to obtain aroma or other bioproducts.
433. V. Popova, T. Ivanova, A. Stoyanova, V. Nikolova, T. Hristeva, and V. D. Zheljazkov, “GC-MS Composition and Olfactory Profile of Concretes from the Flowers of Four Nicotiana Species,” Molecules, vol. 25, no. 11, p. 2617, Jan. 2020. doi: 10.3390/molecules25112617. Abstract
     The genus Nicotiana (Solanaceae) includes over 70 species, with a long history of traditional use; many of them are nowadays used in bioengineering, biosynthesis, molecular biology, and other studies, while common tobacco, N. tabacum L., is one of the most economically important industrial crops worldwide. Although Nicotiana species have been extensively investigated, relatively less research has focused on flowers, especially research related to obtaining aromatic products for cosmetic and perfumery use. On the other hand, there is evidence that Nicotiana flowers accumulate various secondary metabolites with a distinct aroma and biological activities, and the flowers represent a biomass available in sufficient quantities. Therefore, this study aimed to determinate the chemical composition (by GC-MS) and the olfactory profiles of a specific type of natural aromatic product (concrete), obtained from the flowers of four Nicotiana species, in a direct comparison between them. The yields of extracted concrete were sufficiently high, varying between the species, 1.4% (N. rustica L.), 2.5% (N. glutinosa L.), 1.6% (N. alata Link&Otto genotype with white flowers), 2.7% (N. alata genotype with pink flowers), 3.2% (N. tabacum, Oriental type), and 5.2% (N. tabacum, Virginia type). The major components of the obtained concretes belonged to different chemical classes: N. rustica and N. tabacum (OR), the hydrocarbons n-tetratriacontane (14.5%; 15.0%) and n-triacontane (12.1%; 13.3%), and 3-methyl-pentanoic acid (11.1%; 12.2%); N. glutinosa, the diterpenes sclareol (25.9%), 3-α-hydroxy-manool (16.3%), and 13-epimanool (14.9%); N. alata (WF), the phenylpropanoid terephthalic acid and di(2-ethylhexyl) ester (42.9%); N. alata (PF), the diterpene tributyl acetylcitrate (30.7%); and N. tabacum (FCV), the hydrocarbons n-hexacosane (12.9%) and n-pentacosane (12.9%). Each of the flower concretes revealed a characteristic odor profile. This is the first report about Nicotiana species as a source for obtaining flower concretes; these initial results about the concrete yield, olfactory profile, and chemical composition are a prerequisite for the possible processing of Nicotiana flowers into new aromatic products for use in perfumery and cosmetics. The study provides new data in favor of the potential of the four Nicotiana species as aromatic plants, as well as a possible alternative use of flowers, a valuable, but discarded, plant material in other applications.
434. F. B. Power and A. H. Salway, “LIX.—The Constituents of Withania Somnifera,” Journal of the Chemical Society, Transactions, vol. 99, no. INVALID_SCITE_VALUE, pp. 490–507, Jan. 1911. doi: 10.1039/CT9119900490. Abstract
     No abstract available
435. R. B. Prajapati, K. M. Gediya, and D. M. Patel, “Feasibility of Vegetable Intercropping in Rustica Tobacco (Nicotiana Rustica L.) Under Middle Gujarat Conditions,” Advances in Life Sciences, p. 6, 2016. Abstract
     A field experiment was conducted at Bidi Tobacco Research Station, Anand Agricultural University, Anand, Gujarat during rabi season of 2013-14. The experiment consisted of ten treatments comprising of tobacco + vegetable crops like T1(TobaccoAlone), T2 (Tobacco + Cabbage, 1:2), T3 (Tobacco + Cauliflower, 1:2), T4 (Tobacco + Onion, 1:2), T5 (Tobacco + Garlic, 1:3), T6 (Tobacco + Radish, 1:3), T7 (Tobacco + Beet, 1:3), T8 (Tobacco + Fennugreek, 1:9), T9 (Tobacco + Spinach, 1:3) and T10 (Tobacco + Coriander, 1:3) was tried out in a RBD with four replications. Cured leaf yield (3633 kg ha-1) of tobacco was significantly higher with tobacco + garlic over sole tobacco. Tobacco equivalent yield (7762 kg ha-1) was significantly the highest with tobacco + spinach over sole tobacco. Increase in tobacco equivalent yield was 154 % over rest of the treatments including sole tobacco. Net return ( 213678 ha-1) was higher with tobacco + spinach with 1:2.9 BCR ratio. It was concluded that higher tobacco equivalent yield and net returns could be achieved with tobacco + spinach, an economical viable intercropping system for tobacco belt of Middle Gujarat.
436. T. Pramodkumar, M. Shiragur, and B. Chandrakant, “Effect of Bio-Inoculants on Growth, Dry Root Yield and Quality in Ashwagandha (Withania Somnifera L. Dunal.),” International Journal of Current Microbiology and Applied Sciences, vol. 7, no. 09, pp. 2203–2212, Sep. 2018. doi: 10.20546/ijcmas.2018.709.272.
437. C. W. Prather and E. C. Sisler, “Glycine and Glyoxylate Decarboxylation in Nicotiana Rustica Roots,” Phytochemistry, vol. 11, no. 5, pp. 1637–1647, May 1972. doi: 10.1016/0031-9422(72)85011-8. Abstract
     Glycine was decarboxylated only by intact mitochondria to yield carbon dioxide, formaldehyde, and ammonia, probably present as pyridoxamine phosphate. The formaldehyde could become incorporated into serine, via N5N10 methylene-FH4, and a requirement was demonstrated for pyridoxal phosphate. Similarly, glyoxylate with pyridoxamine phosphate was also decarboxylated to formaldehyde and carbon dioxide. Glyoxylate could be decarboxylated by at least two additional pathways. One consisted of oxidative decarboxylation yielding formate and carbon dioxide, and requiring thiamine pyrophosphate, manganese ions, and oxygen. The other consisted of glyoxylate condensation with 2-oxoglutarate, yielding carbon dioxide and an intermediate which, upon decarboxylation, appeared to be hydroxylevulinic acid.
438. N. Praveen, P. M. Naik, S. H. Manohar, and H. N. Murthy, “Distribution of Withanolide a Content in Various Organs of Withania Somnifera (L.) Dunal.,” International Journal of Pharma and Bio Sciences, vol. 1, no. 3, 2010. https://www.cabdirect.org/cabdirect/abstract/20113372347. Abstract
     Withanolide A is an important secondary metabolite in Withania somnifera, which is having a high medicinal value and possesses potent anti-tumor and antioxidant properties. Distribution of withanolide A in various organs of Withania somnifera was investigated by High Performance Liquid Chromatography (HPLC) method. The quantitative distribution of withanolide A was different in various organs...
439. S. U. Qadir, V. Raja, W. A. Siddiqui, M. N. Alyemeni, and P. Ahmad, “Foliar Concentrations of Selected Elements, Assessment of Oxidative Stress Markers and Role of Antioxidant Defense System Is Associated with Fly Ash Stress Tolerance in Withania Somnifera,” Journal of Plant Growth Regulation, vol. 40, no. 4, pp. 1450–1465, Aug. 2021. doi: 10.1007/s00344-020-10200-6. Abstract
     Increased dependence on thermal power has resulted in a significant increase in the generation of fly ash (FA), which exacerbates environmental pollution. In order to mitigate this source of pollution, we propose covering FA dumps with a layer of planted vegetation. Due to varying degrees of tolerance and their sessile nature, plants are themselves susceptible to stress from pollution. This suggests that an investigation to assess the role of wild growing plants for management of FA dumps, where selected wild plants could be grown to mitigate consequences of FA. The present study assesses oxidative damage and the foliar concentration of metals in Withania somnifera growing wild at the Badarpur Thermal Power Plant (BTPP) compared to those growing at a control site. Plants growing at the BTPP showed significantly higher foliar concentrations of Pb, Mn, and Fe, and low concentrations of Ni and Cd. The plants at the BTPP site showed signs of oxidative stress as indicated by enhanced levels of malondialdehyde and electrolyte leakage from cells. The CO2 assimilation rate, net photosynthetic rate, rate of transpiration, stomatal conductance decrease, while water use efficiency, and air pollution tolerance index increase. Among air pollution tolerance index parameters, relative water content showed a significant increase with FA pollution stress at the BTPP. A significant decrease was observed in leaf morphology single leaf area, leaf length, and leaf width and biochemical parameters (Chlorophyll a, Chlorophyll b, total chlorophyll, and carotenoids. Moreover, FA pollution stress induces oxidative stress in W. somnifera through a significant and enhanced production of reactive oxygen species (ROS). According to our observations, the ability of W. somnifera to effectively coordinate superoxide dismutase, ascorbate peroxidase, and glutathione reductase activities involved in the scavenging of ROS along with the enhanced increment of nonenzyme activities (total ascorbic acid, proline, and oxidized glutathione) could be related to FA stress tolerance in W. somnifera.
440. A. C. Qamaruddin Jogi, N. A. P. Naseeruddin, and A. H. J. and N. Babar, “38. Impact of Phyto-Hormone Concentrations in Optimizing Cell Suspension Culture of Flue-Cured Tobacco (Nicotiana Tabaccum L.) Cultivars,” Pure and Applied Biology (PAB), vol. 9, no. 4, pp. 2589–2598, Oct. 2020. https://mail.thepab.org/index.php/journal/article/view/1619. Abstract
     Plant cell suspension cultures are mostly used for the biochemical investigation of cell physiology, growth, metabolism and production of secondary metabolites. The study was undertaken to optimize protocol for callus induction and cell suspension in K-326 and Honghuadajinyuan (HD) cultivars of tobacco (Nicotiana tabaccum). The experiment was conducted at the Key Laboratory of Tobacco Genetic Improvement and Biotechnology, Chinese Academy of Agricultural Sciences (CAAS), Qingdao, P.R. China. In this study explants leaf were inoculated on the (Murashig and Skoog) media with supplementation of exogenous growth regulators of plant. For the morphogenic callus proliferation and indication form explants, (MS) medium was used with (NAA, 6-BA and 2, 4-D phytohormones. Callus derived leaves were stabilized for 4 week of period. The tobacco cell suspension culture was established initially through the culture of friable leaf derived callus in the medium of liquid callus induction. The results of growth regulator combination significantly (P<.0.05) affected on the calli growth. In this study between different treatments, the highest frequency of callus induction was recorded in the level (1) (i.e. NAA) (.33mg/l) and 2, 4-D (0.5mg/L, followed by the level of 03 (i.e. 0.75mg/LNAA, 2.4mg/L 6-BA and 0.5mg/L2, 4-D), respectively. The subsequently sub-culturing of friable callus on callus induction media enhanced callus biomass subculture cycle. The callus induction obtained from HD and K-326 leaf explants was 82.5% and 80%, respectively. The cell growth curve showed that, cells of HD and K-326 produced highest fresh cell mass of 38.07 and 37.67 g, respectively. The production of callus biomass became stable after three subculture cycles. The cells subsequently grew healthy and maintained well in MS liquid medium supplement with the optimal hormone. The browning occurred when the cultures reached the highest fresh cell mass. Therefore, in order to maintain healthy cultures, sub-culturing should be done before browning occurs. Keywords: Callus induction; Cell culture; Tobacco; Growth regulators http://dx.doi.org/10.19045/bspab.2020.90275
441. R. K. Sharma, “Evaluation of Antioxidant Activities of Withania Somnifera Leaves Growing in Natural Habitats of North-West Himalaya, India,” Journal of Medicinal Plants Research, vol. 6, no. 5, Feb. 2012. doi: 10.5897/JMPR11.257. Abstract
     Evaluation of antioxidant properties of medicinal plants from Indian Himalayan region has been very rarely carried out. Withania somnifera L. is one of the commercially available and most preferred medicinal plants in the Himalayan region due to its aphrodisiac property and potential to cure various diseases. The review of literature has indicated that the antioxidant activities of W. somnifera have not been carried out for so long. Therefore, in the present study an attempt was made to evaluate the antioxidant properties of W. somnifera collected from two different habitats that is, forest and roadside at Kullu, north-west Himalaya. The total phenolic and flavonoid contents and DPPH (1, 1-Diphenyl-2pycrylhydrazyl) scavenging potential of leaves extract of W. somnifera varied significantly between the habitat (p<0.05). The results further showed that the DPPH scavenging potential of leaves extract at forest site was found significantly higher that is 51% over the roadside. The study indicates that antioxidant activities of leaves of W. somnifera L. varies habitat to habitat and antioxidant properties decrease if plants are exposed to vehicular pollution. Therefore, plantation and cultivation of this species in vehicular pollution free areas are suggested.
442. R. A. Raguso, R. A. Levin, S. E. Foose, M. W. Holmberg, and L. A. McDade, “Fragrance Chemistry, Nocturnal Rhythms and Pollination ‘Syndromes’ in Nicotiana,” Phytochemistry, vol. 63, no. 3, pp. 265–284, Jun. 2003. doi: 10.1016/S0031-9422(03)00113-4. Abstract
     GC–MS analyses of nocturnal and diurnal floral volatiles from nine tobacco species (Nicotiana; Solanaceae) resulted in the identification of 125 volatiles, including mono- and sesquiterpenoids, benzenoid and aliphatic alcohols, aldehydes and esters. Fragrance chemistry was species-specific during nocturnal emissions, whereas odors emitted diurnally were less distinct. All species emitted greater amounts of fragrance at night, regardless of pollinator affinity. However, these species differed markedly in odor complexity and emission rates, even among close relatives. Species-specific differences in emission rates per flower and per unit fresh or dry flower mass were significantly correlated; fragrance differences between species were not greatly affected by different forms of standardization. Flowers of hawkmoth-pollinated species emitted nitrogenous aldoximes and benzenoid esters on nocturnal rhythms. Four Nicotiana species in section Alatae sensu strictu have flowers that emit large amounts of 1,8 cineole, with smaller amounts of monoterpene hydrocarbons and α-terpineol on a nocturnal rhythm. This pattern suggests the activity of a single biosynthetic enzyme (1,8 cineole synthase) with major and minor products; however, several terpene synthase enzymes could contribute to total monoterpene emissions. Our analyses, combined with other studies of tobacco volatiles, suggest that phenotypic fragrance variation in Nicotiana is shaped by pollinator- and herbivore-mediated selection, biosynthetic pathway dynamics and shared evolutionary history.
443. M. Rai, D. Acharya, A. Singh, and A. Varma, “Positive Growth Responses of the Medicinal Plants Spilanthes Calva and Withania Somnifera to Inoculation by Piriformospora Indica in a Field Trial,” Mycorrhiza, vol. 11, no. 3, pp. 123–128, Aug. 2001. doi: 10.1007/s005720100115. Abstract
     The medicinal plants Spilanthes calva and Withania somnifera were inoculated with Piriformospora indica, a plant growth-promoting root endophyte, in nurseries and subsequently transferred to the field. A significant increase in growth and yield of both plant species was recorded relative to uninoculated controls. Shoot and root length, biomass, basal stem, leaf area, overall size, number of inflorescences and flowers and seed production were all enhanced in the presence of the fungus. Net primary productivity was also higher than in control plants. The results clearly indicate the commercial potential of P. indica for large-scale cultivation of S. calva and W. somnifera.
444. S. Rajaeian, A. A. Ehsanpour, and M. A. Toghyani, “Changes in Phenolic Compound, TAL, PAL Activity of Nicotiana Rustica Triggered by Ethanolamine Pretreatment under in Vitro Salt Stress Condition,” Iranian Journal of Plant Biology, vol. 7, no. 26, pp. 1–12, Dec. 2015. https://ijpb.ui.ac.ir/article_18984.html. Abstract
     Salt stress is one of the most important factors that endanger plant growth and production all around the world. Application of compounds which increase plant resistance to salt stress obviously has significant importance in reduction of economic losses. One of these compounds is the biological alcohol called ethanolamine, and the present study aimed to understand the way by which ethanolamine exerts its protecting effect against salt stress. For this purpose, 4-week-old Nicotiana rustica plants which had been kept in MS medium were pretreated with ethanolamine and 2 days later they were transferred to MS medium supplemented with 200 mM NaCl for 3 weeks. The our results revealed that Nicotiana rustica plants pretreated with exogenous ethanolamine showed elevated level of TAL activity and anthocyanin content whereas PAL activity was decreased, and reduced level of MDA, total flavonoid and phenol content in plants under salt stress condition. According to these results, it can be suggested that ethanolamine exerts its protective effect by anthocyanin content increase.
445. S. Rajaeian, A. A. Ehsanpour, M. Javadi, and B. Shojaee, “Ethanolamine Induced Modification in Glycine Betaine and Proline Metabolism in Nicotiana Rustica under Salt Stress,” Biologia Plantarum, vol. 61, no. 4, pp. 797–800, Dec. 2017. doi: 10.1007/s10535-017-0704-0. Abstract
     The present study aimed to investigate the effects of ethanolamine on glycine betaine and proline metabolism in Nicotiana rustica under salt stress. The in vitro grown tobacco (Nicotiana rustica) plants were pretreated with ethanolamine (at concentrations 70, 130, 270, and 530 μM for biochemical analysis and only at the concentration of 530 μM for molecular analysis) and then transferred to Murashige and Skoog medium containing 200 mM NaCl for 3 weeks. Our results showed that ethanolamine promoted glycine betaine biosynthesis by an increase in betaine aldehyde dehydrogenase (BADH) gene expression and BADH enzymatic activity. Moreover, ethanolamine pretreatment possibly reduced proline content in salt stressed plants via its negative effect on Δ-pyrroline-5-carboxylate synthase (P5CS) gene expression and P5CS enzymatic activity and its positive effect on proline dehydrogenase (PDH) gene expression and PDH activity.
446. S. O. Rajaeian and A. A. Ehsanpour, “Physiological Responses of Tobacco Plants (Nicotiana Rustica) Pretreated with Ethanolamine to Salt Stress,” Russian Journal of Plant Physiology, vol. 62, no. 2, pp. 246–252, Mar. 2015. doi: 10.1134/S1021443715020156. Abstract
     Plant exposure to stress results in the decomposition of their cell membrane phospholipids, and therefore it can elevate the level of EA (ethanolamine) in the cell, and this elevated level of EA induces an alarm response that activates cellular resistance and tolerance mechanisms. In the present study, in vitro cultured tobacco plants (Nicotiana rustica L.) were pretreated with ethanolamine (EA) before salt treatment. After 3 weeks of salt treatment (200 mM NaCl), the plants pretreated with exogenous EA showed the elevated levels of SOD, CAT and APX activity compared with unpretreated plants. Furthermore, total antioxidant capacity, fresh and dry weight and the content of photosynthetic pigments were also increased. In contrast, H2O2 content decreased under similar conditions. According to the results of this study, it can be suggested that EA pretreatment increased salt tolerance of tobacco plants at least partly by stimulation of antioxidative responses.
447. B. R. Rajeswara Rao, \relax R. A. J. P. U. T. D.K., and \relax A. D. I. G, “Opportunities and Challenges in the Cultivation of Ashwagandha Withania Somnifera (L.) Dunal,” Journal of Pharmacognosy, vol. 3, pp. 88–91, Jan. 2012. Abstract
     Winter cherry or Ashwagandha (Withania somnifera) is an important medicinal plant native to India. The roots are widely used in Indian systems of medicine for more than 4000 years. The roots are credited with several medicinal properties. Recent research suggests phytochemical withaferin A present in the leaves possess antitumor activity. It is drought tolerant annual, hence is cultivated under rainfed condition in marginal soils by small and marginal farmers of Madhya Pradesh, Rajasthan, Andhra Pradesh, Karnataka and other Indian states. The ease of cultivation and high price for the roots is attracting farmers for large scale cultivation. In addition to the roots, leaves and seeds are also marketed enhancing the profits of the farmers. The future is excellent for extending its cultivation and production of numerous value added products from its roots.
448. A. Ramachandran and M. S. Kumar, “FT-IR, UV and Antimicrobial Activity with Ania Somnifera and Withania Obtusifolia.,” International Journal of Pharma and Bio Sciences, vol. 5, no. 4, 2014. https://www.cabdirect.org/globalhealth/abstract/20153064150. Abstract
     The present study was to compare the Phytochemical and Antimicrobial analysis of W. somnifera and W. obtusifolia. In the Ethanolic leaf and root extract of Withania species were screened for phytochemical compounds by qualitative and FT-IR, UV method. Withania species have been used for remedies of human pathogen because they contain therapeutic compounds. This research work was mainly focused to...
449. R. Ramesh Kumar, L. Prasanna Anjaneya Reddy, A. Niranjana Kumar, K. Komaraiah, S. Purnanand, and K. P. Sastry, “Root Textural Quality in Ashwagandha (Withania Somnifera) as Influenced by Crop Growth Periods and Morphotypes,” Industrial Crops and Products, vol. 34, no. 1, pp. 1231–1234, Jul. 2011. doi: 10.1016/j.indcrop.2011.03.016. Abstract
     Ashwagandha (Withania somnifera) is a commercially important medicinal crop and roots are the main portion of the plant used therapeutically. The root textural quality determined by high starch–fiber ratio fetches premium price for the produce. Influence of morphological variants and crop growth intervals on accumulation of starch and crude fiber in roots was studied to assess morphotypical differences and to determine the right harvest stage for high root yield with better physical quality. The morphotypes showed similar trend for dry root yield, whereas differences were observed for starch and crude fiber accumulation. The pattern of starch and fiber accumulation varied with different crop growth periods. In general, crude fiber content increased with the crop duration, whereas starch content and starch–fiber ratio followed a trend of decrease–increase–decrease as influenced by different phenophases. Discussions on suitable harvest stage based on dry root yield and starch–fiber ratio are made.
450. S. Rana et al., “A 12-Deoxywithastramonolide-Rich Somaclonal Variant in Withania Somnifera (L.) Dunal—Molecular Cytogenetic Analysis and Significance as a Chemotypic Resource,” In Vitro Cellular & Developmental Biology - Plant, vol. 48, no. 5, pp. 546–554, Oct. 2012. doi: 10.1007/s11627-012-9458-8. Abstract
     Withania somnifera, commonly known as ashwagandha or Indian ginseng, is a valuable medicinal plant, synthesizing a wide array of pharmacologically active secondary metabolites known as withanolides. In this study, we investigated variation among 54 regenerated plants attained through indirect organogenesis from leaf explants. Organogenic calli were induced on Murashige and Skoog medium containing 2 mg l−1 kinetin and 1 mg l−1 indole-3-butyric acid. High-performance liquid chromatography was used for quantitative determination of the major withanolides in the somaclones. One somaclone (WS-R-1) showed significantly higher accumulation of 12-deoxywithastramonolide (WS-12D; 0.516%) compared to the explant donor mother plant (0.002%). The incidence of somaclonal variation at the cytological level was investigated by studying mitosis and meiosis in relation to chromosome number and structural organization. There were no alterations in chromosome phenotypes, somatic chromosome count, or meiotic behavior. Fidelity at genomic level was evaluated by random amplification of polymorphic DNA (RAPD) analyses, which revealed multiple genetic polymorphisms between the WS-12D over-producing somaclone and the explant donor mother plant. This study demonstrates the capability of inducing chemotypic variability for the development of high-yielding clones due to molecular instability in W. somnifera using an in vitro approach.
451. S. Rana et al., “Molecular Characterization of Two A-Type P450s, WsCYP98A and WsCYP76A from Withania Somnifera (L.) Dunal: Expression Analysis and Withanolide Accumulation in Response to Exogenous Elicitations,” BMC Biotechnology, vol. 14, no. 1, p. 89, Nov. 2014. doi: 10.1186/s12896-014-0089-5. Abstract
     Pharmacological investigations position withanolides as important bioactive molecules demanding their enhanced production. Therefore, one of the pivotal aims has been to gain knowledge about complete biosynthesis of withanolides in terms of enzymatic and regulatory genes of the pathway. However, the pathway remains elusive at the molecular level. P450s monooxygenases play a crucial role in secondary metabolism and predominantly help in functionalizing molecule core structures including withanolides.
452. G. Rani, G. S. Virk, and A. Nagpal, “Somatic Embryogenesis in Withania Somnifera (L.) Dunal,” Journal of Plant Biotechnology, vol. 6, no. 2, pp. 113–118, 2004. https://koreascience.kr/article/JAKO200412910492656.page. Abstract
     Somatic embryos were formed from calli obtained from axillary shoots (raised from nodal segments of glasshouse-grown plants under aseptic conditions), internodal segments (from in vitro-raised plants), and root and coty-ledonary leaf segments (from in vitro-raised seedlings) after 8 weeks of initial culture. Embryo formation was the highest (97.33%) from cotyledonary leaf callus on Mura-shige and Skoog’s (MS) medium containing kinetin (KN) (3 mg/L). Somatic embryo induction was lesser with different combinations of auxins while it increased to 100% in internodal segment and cotyledonary leaf calli with 6-benzyladenine (BA) (2mg/L) along with 2,3,5-triiodobenzoic acid (TIBA) (2mg/L). The shoots were induced from somatic embryos raised from root, coty-ledonary leaf and internodal segment calli grown on MS medium containing BA in combination with indole-3-acetic acid (IAA). Maximum of 66.67% cultures formed shoots on MS medium containing BA (1mg/L) in combination with IAA (2mg/L). The shoots raised from somatic embryos were rooted on MS medium supplemented with indole-3-butyric acid (IBA) (2mg/L). The plantlets transferred to the field showed 70% survival rate after one year.
453. G. Rani and I. S. Grover, “In Vitro Callus Induction and Regeneration Studies in Withania Somnifera,” Plant Cell, Tissue and Organ Culture, vol. 57, no. 1, pp. 23–27, Apr. 1999. doi: 10.1023/A:1006329532561. Abstract
     Callus cultures were initiated from axillary leaves, axillary shoots, hypocotyls, and root segments on Murashige and Skoog (MS) (1962) medium supplemented with 2,4-D (2 mg l−1) and KN (0.2 mg l−1). Shoots differentiated best from axillary shoot base callus on MS medium containing BA (2 mg l−1). Regenerated shoots rooted best on MS medium containing IBA (2 mg l−1) alone, and IBA (2 mg l−1) with IAA (2 mg l−1). Plantlets were transferred to pots containing sand and soil mixture, acclimatized in a culture room and afterwards transferred to the glasshouse.
454. A. Rani, M. Kumar, and S. Kumar, “In Vitro Propagation of Withania Somnifera (L.) Dunal. from Shoot Apex Explants,” Journal of Applied and Natural Science, vol. 6, no. 1, pp. 159–163, Jun. 2014. doi: 10.31018/jans.v6i1.393. Abstract
     In vitro rapid micropropagation of Withania somnifera; has been achieved using shoot apex explants, when cultured on MS medium with different auxins: Indole Butyric Acid (IBA) 0.5-4 mg/l, Indole Acetic Acid (IAA) 2 mg/l and Naphthalene Acetic Acid (NAA) 0.5-3 mg/l and cytokinin: Benzyl Amino Purine (BAP) 0.5-4 mg/l. NAA and BAP stimulated direct as well as callus mediated multiple shoot. IBA alone and in combination with IAA stimulated rhizogenesis, while IBA with NAA proliferated callus-mediated rhizogenesis. The rooted plantlets through sequential hardening process could be established in the field.
455. P. S. Rao and N. S. Rangaswamy, “In Vitro Development of the Pollinated Pistils of Nicotiana Rustica L.,” Botanical Gazette, vol. 133, no. 4, pp. 350–355, Dec. 1972. doi: 10.1086/336654. Abstract
     In the self-incompatible species Nicotiana rustica attempts were made to overcome the incompatibility barrier through culture of ovule and pollen, but these were unsuccessful. On the contrary, in cultures of entire pistils which were cross-pollinated in vitro, the process of fertilization and the development of embryo and endosperm proceeded normally. The physiology of fruit growth in vitro and in vivo and the role of calyx in fruit development were investigated. It is suggested that the pistil culture technique may prove useful in overcoming certain prefertilization barriers to incompatibility, particularly the precocious abscission of the ovary.
456. C. D. Raper Jr. and W. T. Smith, “Factors Affecting the Development of Flue-Cured Tobacco Grown in Artificial Environments. V. Effects of Humidity and Nitrogen Nutrition1,” Agronomy Journal, vol. 67, no. 3, pp. 307–312, 1975. doi: 10.2134/agronj1975.00021962006700030007x. Abstract
     Controlled environment rooms (CERs) were used to investigate effects of relative humidity on physical and chemical properties of mature, flue-cured tobacco (Nicotiana tabacum L.) leaves. The relative humidities in separate CERs were monitored with a dewpoint hygrometer and maintained at 90% (high) or 65% (low) relative humidity. Plants were moved between the two CERs to evaluate effects of diurnal and seasonal variations in relative humidity. The apparent effects of humidity on leaf size and shape were dependent on age of the plants. For plants grown in continuous high humidity, leaf sizes were greater at lower and middle stalk positions, but were less at upper stalk positions, than for plants grown in continuous low humidity. The reduction of areas of upper leaves by high humidity was a residual response to the humidity during the initial 3 to 5 weeks after transplanting. Initial periods of high humidity, whether followed by low or high humidity, also resulted in abnormally elongated upper leaves. Furthermore, variations in total plant leaf areas which resulted from diurnal variations in humidity were highly correlated with total nitrogen content in leaf tissues. These results suggest that suppression of nitrogen uptake by high humidity during a critical period of leaf formation was directly involved. Humidity during later stages of growth apparently has effects on the chemical balance within tobacco plants unassociated with nitrogen accumulation. High humidity during the light period tended to increase both starch and alkaloid composition of leaves.
457. B. Rathod, D. Chaudhari, J. Patel, S. K. Patel, and R. Patel, “Correlation and Path Analysis Studies in Rustica Tobacco (Nicotiana Rustica L.),” Jan. 2021. doi: 10.18782/2582-2845.8787. Abstract
     The present investigation was carried out at Agricultural Research Station, Sardarkrushinagar Dantiwada Agricultural University, Ladol during rabi 2019-20. The experimental material consisted of thirty-five genotypes which were evaluated in Randomized Block Design with four replications. The genotypes were evaluated for 11 characters. Correlation studies revealed that cured leaf yield per plant had positive and significant association with plant height, number of leaves per plant, leaf length, leaf width at both genotypic and phenotypic levels. Path coefficients analysis based on genotypic correlation revealed that number of leaves per plant showed highest positive direct effect followed by leaf width, days to flowering, plant height, reducing sugar content and leaf length. Path coefficients analysis based on genotypic correlation revealed that number of leaves per plant showed highest positive direct effect followed by leaf width, days to flowering, plant height, reducing sugar content and leaf length. Hence, these traits may be directly attributed for the improvement of cured leaf yield and important in the selection of better genotypes in rustica tobacco. Hence, these traits may be directly attributed for the improvement of cured leaf yield and important in the selection of better genotypes in rustica tobacco. To improve cured leaf yield, proper attention should therefore be paid to number of leaves per plant, leaf length, leaf width, days to flowering and plant height. Selection for these traits will therefore directly become helpful in increasing the cured leaf yield in rustica tobacco.
458. S. Ray and S. Jha, “Production of Withaferin A in Shoot Cultures of Withania Somnifera,” Planta Medica, vol. 67, no. 5, pp. 432–436, 2001. doi: 10.1055/s-2001-15811. Abstract
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459. S. Ray, B. Ghosh, S. Sen, and S. Jha, “Withanolide Production by Root Cultures of Withania Somnifera Transformed with Agrobacterium Rhizogenes,” Planta Medica, vol. 62, no. 6, pp. 571–573, Dec. 1996. doi: 10.1055/s-2006-957977. Abstract
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460. S. Ray and S. Jha, “Withanolide Synthesis in Cultures of Withania Somnifera Transformed with Agrobacterium Tumefaciens,” Plant Science, vol. 146, no. 1, pp. 1–7, Aug. 1999. doi: 10.1016/S0168-9452(99)00077-1. Abstract
     Transformed organ cultures of Withania somnifera were established following infection with wild type nopaline and octopine strains of Agrobacterium tumefaciens. The oncogenic strains had different levels of virulence on two genotypes studied, the main difference was found in the nature of the galls formed and in their subsequent morphological competence. Ten percent of the galls obtained following infection with nopaline strain N2/73 spontaneously developed shooty teratomas of altered phenotype. The shooty teratomas grew in unsupplemented basal medium and were able to synthesize both the major withanolides of the parent plants. Withanolide synthesis in shooty teratomas was much higher (0.07–0.1% withaferin A and 0.085–0.025% withanolide D) than in non-transformed shoot cultures.
461. M. J. C. Rhodes, M. Hilton, A. J. Parr, J. D. Hamill, and R. J. Robins, “Nicotine Production by ‘Hairy Root’ Cultures of Nicotiana Rustica: Fermentation and Product Recovery,” Biotechnology Letters, vol. 8, no. 6, pp. 415–420, Jun. 1986. doi: 10.1007/BF01026743. Abstract
     The production of nicotine by cultures ofNicotiana rustica transformed withAgrobacterium rhizogenes has been examined in a packed bed fermenter as a two-stage batch/continuous-flow system. A substantial proportion of the nicotine synthesised in the batch phase may be subsequently harvested from the medium. The possibility of improving product recovery using macroreticular adsorbents is considered.
462. P. Rita and A. Datta, “An Updated Overview on Atropa Belladonna L,” International Research Journal of Pharmacy, vol. 2, pp. 11–17, Nov. 2011. Abstract
     Atropa belladonna L. (Family: Solanaceae; commonly known as belladonna, deadly nightshade, devil’s berries amongst others), a perennial herb (2n=72) is native of Europe, North Africa and Western Asia, possesses a long tradition as one of the classic poisons of antiquity. The species is also the source of atropine alkaloid (dl-hyoscyamine) and is important in the study of autonomic pharmacology. Considering the therapeutic uses of A. belladonna as well as its significant toxic effects (due to tropane alkaloids including scopolamine and hyoscyamine), an overview on all necessary aspects is documented to provide information for further exploration of the species for human benefits.
463. R. J. Robins, J. D. Hamill, A. J. Parr, K. Smith, N. J. Walton, and M. J. C. Rhodes, “Potential for Use of Nicotinic Acid as a Selective Agent for Isolation of High Nicotine-Producing Lines of Nicotiana Rustica Hairy Root Cultures,” Plant Cell Reports, vol. 6, no. 2, pp. 122–126, Apr. 1987. doi: 10.1007/BF00276668. Abstract
     The addition of exogenous nicotinic acid, nicotinamide or nicotine was studied with reference to their effects on growth and alkaloid production by hairy root cultures of Nicotiana rustica. Nicotinic acid and nicotinamide were toxic (50% phytostatic dose being 2.4 and 9 mM respectively) while nicotine was not toxic below 10 mM. Nicotinic acid (up to 5 mM) was found to be phytostatic rather than phytotoxic. Roots exposed to increasing nicotinic acid or nicotinamide levels had altered alkaloid accumulation patterns relative to the controls. The principal effects were to increase the intracellular and extracellular levels of anatabine and nicotine, with a markedly greater proportion of anatabine being produced. The use of nicotinic acid as a selection agent for the recovery of higher alkaloid-producing lines is identified and discussed.
464. W. Röper, M. Schulz, E. Chaouiche, and K. A. Meloh, “Nicotine Production by Tissue Cultures of Tobacco as Influenced by Various Culture Parameters,” Journal of Plant Physiology, vol. 118, no. 5, pp. 463–470, Apr. 1985. doi: 10.1016/S0176-1617(85)80206-6. Abstract
     Tissue culture systems of tobacco (Nicotiana tabacum L., Nicotiana rustica L.) are described, in which up to 5% and more nicotine accumulated on a dry weight basis. Maximum amounts of nicotine (5.3 %, yield 920 mg/1) were detected in mixotrophic green cell suspensions of a newly developed flue-cured tobacco cultivar (N. tabacum cv. Delcrest X cv. McNair 133). This cultivar also showed maximum nicotine content compared to other strains when cultured as callus or heterotrophic cell suspension. Nicotine content was increased up to 29-fold (2. 9 %, yield 360 mg/1) compared to the stock cultures, when a two- or three-stage culture was employed by reducing the auxin and cytokinin levels, raising medium sucrose to 40 gil and replacing ammonium nitrate by potassium nitrate in the production medium. Cells in 20 1 batch fermentor cultures accumulated up to 1 % nicotine with a final yield of 28 mg/1, thus producing much less nicotine.
465. J. R. Rout et al., “Biochemical, Molecular, and Elemental Profiling of Withania Somnifera L. with Response to Zinc Stress,” Environmental Science and Pollution Research, vol. 26, no. 4, pp. 4116–4129, Feb. 2019. doi: 10.1007/s11356-018-3926-6. Abstract
     Zn stress seriously induces various toxic responses in Withania somnifera L., when accumulated above the threshold level which was confirmed by investigating the responses of protein, expression of antioxidant enzymes, and elemental profiling on accumulation of Zn. Zn was supplemented in the form of ZnSO4 (0, 25, 50, 100, and 200 μM) through MS liquid medium and allowed to grow the in vitro germinated plants for 7 and 14 days. The study revealed that when the application of Zn increased, a significant reduction of growth characteristics was noticed with alterations of proteins (both disappearance and de novo synthesis). The activity of CAT, SOD, and GPX were increased up to certain concentrations and then declined, which confirmed through in-gel activity under different treatments. RT-PCR was conducted by taking three sets of genes from CAT (RsCat, Catalase1, Cat1) and SOD (SodCp, TaSOD1.2, MnSOD) and found that gene RsCat from CAT and MnSOD from SOD have shown maximum expression of desired genes under Zn stress, which indicate plant’s stress tolerance mechanisms. The proton-induced X-ray emission study confirmed an increasing order of uptake of Zn in plants by suppressing and expressing other elemental constituents which cause metal homeostasis. This study provides insights into molecular mechanisms associated with Zn causing toxicity to plants; however, cellular and subcellular studies are essential to explore molecule-molecule interaction during Zn stress in plants.
466. J. R. Rout, S. S. Ram, R. Das, A. Chakraborty, M. Sudarshan, and S. L. Sahoo, “Copper-Stress Induced Alterations in Protein Profile and Antioxidant Enzymes Activities in the in Vitro Grown Withania Somnifera L.,” Physiology and Molecular Biology of Plants, vol. 19, no. 3, pp. 353–361, Jul. 2013. doi: 10.1007/s12298-013-0167-5. Abstract
     Withania somnifera L. seedlings were grown in half-strength MS (Murashige and Skoog) basal medium for 4 weeks and then transferred to full-strength MS liquid medium for 3 weeks. The sustainable plants were subcultured in the same medium but with different concentrations (0, 25, 50, 100 and 200 μM) of Cu for 7 and 14 days. The growth parameters (root length, shoot length, leaf length and total number of leaves per plant) showed a declining trend in the treated plants in a concentration dependant manner. Roots and leaves were analyzed for protein profiling and antioxidant enzymes [catalase (CAT, EC 1.11.1.6), superoxide dismutase (SOD, EC 1.15.1.1) and guaiacol peroxidase (GPX, EC 1.11.1.7)]. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of crude protein extracts showed the appearance of some new proteins due to Cu treatment. In plant samples grown with 25 and 50 μM of Cu, a rapid increase in antioxidant activities were noticed but at higher concentration (100 and 200 μM) the activities declined. Isoforms of CAT, SOD and GPX were separated using non-denaturing polyacrylamide gel electrophoresis and concentration specific new isoforms were noticed during the study. Isoforms of the antioxidant enzymes synthesized due to Cu stress may be used as biomarkers for other species grown under metal stress.
467. J. R. Rout et al., “Effect of Iron Stress on Withania Somnifera L.: Antioxidant Enzyme Response and Nutrient Elemental Uptake of in Vitro Grown Plants,” Ecotoxicology, vol. 24, no. 2, pp. 401–413, Mar. 2015. doi: 10.1007/s10646-014-1389-1. Abstract
     In the present study the response of antioxidant enzyme activities and the level of expression of their corresponding genes on bioaccumulation of iron (Fe) were investigated. In vitro germinated Withania somnifera L. were grown in Murashige and Skoog’s liquid medium with increasing concentrations (0, 25, 50, 100 and 200 µM) of FeSO4 for 7 and 14 days. Root and leaf tissues analyzed for catalase (CAT, EC 1.11.1.6), superoxide dismutase (SOD, EC 1.15.1.1) and guaiacol peroxidase (GPX, EC 1.11.1.7), have shown an increase in content with respect to exposure time. Isoforms of CAT, SOD and GPX were separated using non-denaturing polyacrylamide gel electrophoresis and observed that the isoenzymes were greatly affected by higher concentrations of Fe. Reverse transcriptase polymerase chain reaction analysis performed by taking three pairs of genes of CAT (RsCat, Catalase1, Cat1) and SOD (SodCp, TaSOD1.2, MnSOD) to find out the differential expression of antioxidant genes under Fe excess. RsCat from CAT and MnSOD from SOD have exhibited high levels of gene expression under Fe stress, which was consistent with the changes of the activity assayed in solution after 7 days of treatment. Analysis by proton induced X-ray emission exhibited an increasing uptake of Fe in plants by suppressing and expressing of other nutrient elements. The results of the present study suggest that higher concentration of Fe causes disturbance in nutrient balance and induces oxidative stress in plant.
468. J. R. Rout and S. L. Sahoo, “Morphological and Protein Profile Alterations in Withania Somnifera l. with Response to Iron Stress,” Indian Journal of Life Sciences, vol. 2, no. 1, pp. 19–24, Dec. 2012. https://go.gale.com/ps/i.do?p=AONE&sw=w&issn=22771743&v=2.1&it=r&id=GALE%7CA357968979&sid=googleScholar&linkaccess=abs. Abstract
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469. S. Roy, S. Amarnath, and P. M. Bhattacharyya, “Screening of Nicotiana Rustica Tobacco Germplasm for Resistance to Hollow Stalk Disease in North Bengal,” Jun. 2012. http://krishi.icar.gov.in/jspui/handle/123456789/50368. Abstract
     Rustica tobacco (Nicotiana rustica) germplasm maintained at Central Tobacco Research InstituteResearch Station, Dinhata (West Bengal) were screened for resistance to hollow stalk disease under artificial inoculation. A total of 182 N. rustica germplasm entries were screened in a phased manner from the year 2007-08 onwards. A scale was considered for disease reaction (soft rot in stem pith) measuring up to 2, 2 - 3 and > 3 cm for resistant, moderately resistant and susceptible types, respectively for cataloguing the entries. In the year 2007-08 a total of 90 entries were screened and 20 were selected based on disease reaction from < 2 cm to 2.05 cm. For confirmation, these 20 entries were again tested in 2008-09 along with 60 new entries. In the year 2009-10, 23 entries were included from the previous year (2008-09) along with 32 new entries. Only accession White Pathar was found to be resistant (soft rot up to 2 cm linear length) consistently for successive three years. Bengthuli, an entry from Assam showed resistant disease reaction for the years 2009 -10 and 201011 only.
470. S. T. S, D. Divakaran, S. S, S. Rebello, and D. J. M. S*, “Antimicrobial and Immunomodulatory Potential of Endophytic Fungus Fusarium Solani Isolated from Withania Somnifera,” World Journal of Pharmaceutical Research, vol. 3, no. 10, pp. 879–890, 2014. https://wjpr.net/abstract_show/1832. Abstract
     Recent research indicate that pharmaceutical compounds produced by medicinal plants can also be produced by their inhabitant endophytes and this paves way to a new methodology of science utilizing endophytes as a source of bioactive compounds. This study aimed to add more information on endophyte driven biodiversity and bioactive potential of Withania. About 29 endophytic fungal isolates belonging to 16 different species were isolated from Withania root segments. Preliminary screening for antibacterial agents against common pathogenic bacteria led to isolation six endophytic fungi viz, F.semitectum, F.avenaceum, Fusarium sp, F.oxysporum, Aspergillus sp and F. redolens. The minimum inhibitory concentration of the extracts ranged in concentrations 21-78μg/ml. Endophytic isolate Fusarium solani also possessed immunomodulatory role and bioactive compounds in its the ethyl acetate extract were analysed by GC-MS. Based on its antibacterial and immunomodulatory potential, Fusarium solani was selected, characterized genotypically by 18S rDNA typing and the sequence was deposited in NCBI database under accession number KJ193849. The current study also describes F.solani as a suitable source for biocompound extraction due to its antibacterial and immunomodulatory role. Works on the mycotic biodiversity of endophytes of Withania and their potential to produce novel bioactive compounds are meager and this work becomes relevant.
471. M. P. S. S, “A Comprehensive Review on Medicinal Herb Withania Somnifera (L.) Dunal in Women’s Health: A Rejuvenator From Siddha Medicine,” Current Pharmacology Reports, vol. 8, no. 1, pp. 72–77, Feb. 2022. doi: 10.1007/s40495-021-00274-5. Abstract
     Medicinal herbs have served mankind since several centuries and hold the rich heritage of treating dreadful diseases. One such potential herb that exists in the siddha system of medicine is Withania somnifera (WS) known by its name Amukkara. Numerous preclinical and clinical studies substantiate the biomedical application of Amukkara, some of which includes anti-cancer, antioxidant, anxiolytic, adaptogen, and anti-inflammatory properties. Despite its extensive efficacy, the herb WS is a well-known remedy for diseases and infections that are specific to women’s health. The main objective of the present review is to prime the health benefits of WS over the health and well-being of women in our society.
472. F. Sabir et al., “Bioconversion of Artemisinin to Its Nonperoxidic Derivative Deoxyartemisinin through Suspension Cultures of Withania Somnifera Dunal,” Zeitschrift für Naturforschung C, vol. 65, no. 9-10, pp. 607–612, Oct. 2010. doi: 10.1515/znc-2010-9-1013. Abstract
     Biotransformation of artemisinin was investigated with two different cell lines of suspension cultures of Withania somnifera. Both cell lines exhibited potential to transform artemisinin into its nonperoxidic analogue, deoxyartemisinin, by eliminating the peroxo bridge of artemisinin. The enzyme involved in the reaction is assumed to be artemisinin peroxidase, and its activity in extracts of W. somnifera leaves was detected. Thus, the non-native cell-free extract of W. somnifera and suspension culture-mediated bioconversion can be a promising tool for further manipulation of pharmaceutical compounds.
473. F. Sabir, S. Mishra, R. S. Sangwan, J. S. Jadaun, and N. S. Sangwan, “Qualitative and Quantitative Variations in Withanolides and Expression of Some Pathway Genes during Different Stages of Morphogenesis in Withania Somnifera Dunal,” Protoplasma, vol. 250, no. 2, pp. 539–549, Apr. 2013. doi: 10.1007/s00709-012-0438-y. Abstract
     Withania somnifera Dunal is an important and extensively studied medicinal plant; however, there is no report available that relates withanolide content and its profile in relation to the expression of pathway genes during different morphogenic stages. In this study, withanolide A, withaferin A, and withanone, the major withanolides of W. somnifera, were measured in different in vitro stages during organogenesis, viz., shoot to root (direct rhizogenesis)/root to shoot (indirect via callus phase) transition vis-à-vis expression levels of key pathway genes involved in withanolide biosynthetic pathways. The morphogenic transitions were found to be tightly linked to the pattern of accumulation of withanolides. The high expression levels of most of the pathway genes in in vitro shoots in comparison to in vitro root and callus tissues exhibited a direct co-relation with the maximum withanolide content (>2.7 mg/gDW). The biogenesis of withaferin A, a major constituent of the leaves, was however found to be tightly linked to shoots/green tissue. In addition, we were also able to establish an efficient regeneration system from roots for their further utilization in biotechnological applications.
474. F. Sabir, R. S. Sangwan, R. Kumar, and N. S. Sangwan, “Salt Stress-Induced Responses in Growth and Metabolism in Callus Cultures and Differentiating In Vitro Shoots of Indian Ginseng (Withania Somnifera Dunal),” Journal of Plant Growth Regulation, vol. 31, no. 4, pp. 537–548, Dec. 2012. doi: 10.1007/s00344-012-9264-x. Abstract
     In vitro-grown shoots and calli of Withania somnifera, an important medicinal plant, were exposed to various types of salts under in vitro culture conditions. Membrane permeability, lipid peroxidation, and the antioxidant system increased in shoots as well as in unorganized callus tissues under all the three concentrations of KCl, NaCl, KNO3, NaNO3, and CaCl2. The growth responses of shoots and callus cultures under various salt treatments revealed that the tissue could grow better under NaCl and KNO3 compared to other salts and the in vitro shoots appeared healthy at 50 mM concentration of NaCl and KNO3. The activity of antioxidant enzymes such as catalase (CAT), ascorbate peroxidase, guaiacol peroxidase, lipoxygenase, polyphenol oxidase, and glutathione reductase increased under salt treatments, especially at higher concentrations. The greatest activity increase was recorded for peroxidases, whereas CAT was the least responsive. Only two isoforms, Mn-superoxide dismutase (Mn-SOD) and Fe-SOD, could be visualized in callus tissue while Cu/Zn-SOD was absent. Diaphorase 4 was totally missing in callus tissue and was detected only in shoots. Phenolics accumulated at all the concentrations of the salts tested as an induced protective response. The higher concentration of CaCl2 produced maximum increases in antioxidants and enzymatic activities compared to other salts. Thus, for W. somnifera the presence of excess calcium in the growing medium is most deleterious compared to other salts. Results also suggest that the nonenzymatic and enzymatic antioxidant systems of both the tissues played a primary role in combating the imposed salt stress.
475. F. Sabir, N. S. Sangwan, N. D. Chaurasiya, L. N. Misra, and R. S. Sangwan, “In Vitro Withanolide Production by Withania Somnifera L. Cultures,” Zeitschrift für Naturforschung C, vol. 63, no. 5-6, pp. 409–412, Jun. 2008. doi: 10.1515/znc-2008-5-616. Abstract
     In vitro multiple shoots, root, callus and cell suspension cultures of Withania somnifera exhibited the potentiality to produce pharmacologically active withanolides. Multiple shoots cultures exhibited an increase in withanolide A accumulation compared to shoots of the mother plant. In vitro generated root cultures as well as callus and suspension cultures also produced withanolides albeit at lower levels.
476. M. Sabour, J. Simmonds, and G. Setterfield, “Variation in Nicotine Content of Cultured Cell Lines of Nicotiana Species and Their Somatic and Sexual Hybrids,” Plant Breeding, vol. 97, no. 4, pp. 324–333, 1986. doi: 10.1111/j.1439-0523.1986.tb01073.x. Abstract
     Several Nicotiana species, their intergeneric sexual and somatic hybrids were compared for their in vitro growth rate and nicotine content. Cell yield on a dry weight basis and HPLC analysis of their nicotine content revealed a very high variation within each cell line from parental, sexual and somatic hybrids for the two traits analyzed. The trigeneríc sexual hybrids (N. rustica × N. debneyi) × N. tabacum and only two of the N. rustica × N. tabacum somatic hybrid lines show a positive heterosis in nicotine content in vitro The in vitro growth rate and nicotine content did not demonstrate a statistically significant correlation.
477. N. Sachan, D. T. Rogers, K.-Y. Yun, J. M. Littleton, and D. L. Falcone, “Reactive Oxygen Species Regulate Alkaloid Metabolism in Undifferentiated N. Tabacum Cells,” Plant Cell Reports, vol. 29, no. 5, pp. 437–448, May 2010. doi: 10.1007/s00299-010-0833-9. Abstract
     Plants produce an immense number of natural products and undifferentiated cells from various plant tissues have long been considered an ideal source for their synthesis. However, undifferentiated plant cells often either lose their biosynthetic capacity over time or exhibit immediate repression of the required pathways once dedifferentiated. In this study, freshly prepared callus tissue was employed to further investigate the regulation of a natural product pathway in undifferentiated tobacco cells. Putrescine N-methyltransferase (PMT) is a pathway-specific enzyme required in nicotinic alkaloid production in Nicotiana species. Callus derived from transgenic Nicotiana tabacum plants harboring PMT promoter–GUS fusions were used to study factors that influence PMT expression. Under normal callus growth conditions in the presence of light and auxin, PMT promoter activity was strongly repressed. Conversely, dark conditions and the absence of auxin were found to upregulate PMT promoter activity, with light being dominant to the repressive effects of auxin. Since reactive oxygen species (ROS) are known by-products of photosynthesis and have been implicated in signaling, their involvement was investigated in transgenic callus by treatment with the ROS scavenger, dimethylthiourea, or catalase. Under highly repressive conditions for alkaloid synthesis, including normal culture conditions in the light, both ROS scavengers resulted in significant induction of PMT promoter activity. Moreover, treatment of callus with catalase resulted in the upregulation of PMT promoter activity and alkaloid accumulation in this tissue. These results suggest that ROS impact the regulation of the alkaloid pathway in undifferentiated cells and have implications for regulation of the pathway in other plant tissues.
478. D. Saco, M. Alvarez, and S. Martín, “Activity of Nitrate Reductase and the Content of Proteins in Nicotiana Rustica Grown with Various Levels of Molybdenum,” Journal of Plant Nutrition, vol. 18, no. 6, pp. 1149–1157, Jun. 1995. doi: 10.1080/01904169509364969. Abstract
     The effect that different levels of molybdenum (11 μMand 111 μM) exert on nitrate reductase (E.C. 1.6.6.1–3) activity (NRA), and the content of total proteins in Nicotiana rustica was studied. Eleven μM and 111 μM of molybdenum increased the activity of nitrate reductase during the growth cycle of N. rustica. Likewise, total protein content (mg/g DW) was increased during the vegetative growth of the plants.
479. S. Saema, L. ur Rahman, R. Singh, A. Niranjan, I. Z. Ahmad, and P. Misra, “Ectopic Overexpression of WsSGTL1, a Sterol Glucosyltransferase Gene in Withania Somnifera, Promotes Growth, Enhances Glycowithanolide and Provides Tolerance to Abiotic and Biotic Stresses,” Plant Cell Reports, vol. 35, no. 1, pp. 195–211, Jan. 2016. doi: 10.1007/s00299-015-1879-5. Abstract
     Overexpression of sterol glycosyltransferase (SGTL1) gene ofWithania somniferashowing its involvement in glycosylation of withanolide that leads to enhanced growth and tolerance to biotic and abiotic stresses.
480. S. Saha, S. B. Modak, and P. K. Basu, “Seed Germination Behavior of Two Varieties of Nicotiana Sp. in Relation to Growth Substances.,” Environment and Ecology, vol. 8, no. 1B, pp. 482–484, 1990. https://www.cabdirect.org/cabdirect/abstract/19900734182. Abstract
     N. rustica and N. tabacum seeds were germinated in 0.001-1000 p.p.m. concn of 4 growth regulators. In N. rustica the greatest increases in percentage germination occurred with 0.01-1.0 p.p.m. IAA, 0.01-0.1 p.p.m. GA and rutin, and 1.0-10.0 p.p.m. quercetin. All treatments increased germination of N. tabacum, with the greatest effects at the lowest concn, except in the case of quercetin. Growth...
481. S. Sahu, A. Prakash, and K. Shende, “Talaromyces Trachyspermus, an Endophyte from Withania Somnifera with Plant Growth Promoting Attributes,” Environmental Sustainability, vol. 2, no. 1, pp. 13–21, Mar. 2019. doi: 10.1007/s42398-019-00045-5. Abstract
     The medicinal plant, Withania somnifera is attributed by valuable medicinal properties and is widely cultivated. It is a need to take care of this plant from synthetic agrochemicals that may be hazardous for health and environment. The aim of the present study was to isolate and screen the endophytic fungi of W. somnifera that  have potential of plant growth promotion and antagonism against plant pathogens. In this study, 22 potential fungal endophytes comprising of species of Alternaria, Aspergillus, Fusarium, Nigrospora, Colletotrichum and Talaromyces identified at National Fungal Culture Collection of India (NFCCI), Pune were isolated. The potential isolate, Talaromyces trachyspermus was confirmed by BLAST and phylogenetic analysis of sequences of rDNA ITS, LSU (D1 D2) and β-tubulin genes. Among all the isolates, T. trachyspermus exhibited comparatively higher activity for hydrolytic enzymes, protease, chitinase, amylase, cellulase and pectinase that are required for antagonistic property. It was observed to be a promising biocontrol agent against plant pathogen, Sclerotinia sclerotiorum. This strain is also characterized with high level of indole acetic acid (IAA), siderophore synthesis, and phosphate solubilization activities that are important for plant growth promotion. This is the first report on endophyte, T. trachyspermus from W. somnifera having potential plant growth promoting traits and biocontrol, which can be further exploited to enhance the medicinal value of the plant.
482. \relax M. A. Saiyem, S. A. Sabur, \relax M. A. Khan, and M. I. Hossain, “Profitability Options of Medicinal Plants Production under Risk: Understanding from Aloe Vera, Bombax Ceiba Root and Withania Somnifera Cultivation in North-West Region, Bangladesh,” Journal of Applied Research on Medicinal and Aromatic Plants, vol. 31, p. 100416, Dec. 2022. doi: 10.1016/j.jarmap.2022.100416. Abstract
     Medicinal plants (MPs) cultivation are an integral part of agricultural diversification, sustainable livelihoods, and profitable options for farmers in Bangladesh. The study is conducted to examine the profitability scenarios of MPs under different risks, i.e. price, yield and market absorption. Primary data were collected from 196 MPs growers of Aloe vera, Bombax ceiba root and Withania somnifera from Natore and Bogura districts of Bangladesh. Monte-Carlo simulation method is applied with considering the embedded risks. Results show that MPs production is profitable alternative to traditional crops in the study area. A. vera production is more profitable compared with B. ceiba root and W. somnifera. The lion share of cost is for human labor followed by seedlings for the selected MPs. Results from Monte-Carlo simulation indicate that there is a significant negative impact on growers’ profits taking into consideration of price, yield and market absorption risks, however, there is a greater possibility of making a profit from A. vera, B. ceiba and W. somnifera production by addressing these risks. The policy should be triggered on taking risk reduction strategies emphasizing more prices and market absorption of MPs by the farmers through establishing contract farming. Farmers should exploit more efficient marketing channels by contracting upstream market actors and avoid traditional long channels that they solely rely on.
483. A. Samad, A. K. Shasany, S. Gupta, P. V. Ajayakuar, M. P. Darokar, and S. P. S. Khanuja, “First Report of a 16SrVI Group Phytoplasma Associated with Witches’-Broom Disease on Withania Somnifera,” Plant Disease, vol. 90, no. 2, pp. 248–248, Feb. 2006. doi: 10.1094/PD-90-0248A. Abstract
     Withania somnifera (L.) Dunal is cultivated in India as an important medicinal cash crop. The whole plant is of great importance in the Indian system of medicine and pharmaceutical industries, but the roots are the main source of active alkaloids. Some of the important alkaloids are tro-pine, pseudotropine, somniferine, colin, withaferin A, withanoides, and a few flavanoides. Typical disease symptoms include phyllody, little leaf, dense clusters of highly proliferating branches with shortened internodes, and resulting witches’-broom. The disease was first observed in and around Lucknow, Uttar Pradesh Province, India during January and February 1992. On the basis of symptoms, transmission electron microscopy (TEM), and antibiotic treatment, the causal organism was identified as a phytoplasma (4). The disease is now spreading to other parts of the country (Gujrat, Haryana, Madhya Pradesh, Punjab, and Rajasthan provinces) with a high disease incidence (70%). In this report, molecular characterization and taxonomic position of the associated phytoplasma is reported. Total genomic DNA was extracted from healthy and infected plants with a modified cetyltrimethylammoniumbromide (CTAB) buffer method. The samples were assayed for the presence of phytoplasma using polymerase chain reaction (PCR) with universal phytoplasma primers P1/P6 (2) for amplification of ribosomal 16S rDNA. PCR product was diluted by 1:200 and used directly as DNA template for nested PCR with primers R16F2n and R16R2 (1). Results showed the presence of an expected 1.5-kb rDNA fragment amplified with the direct PCR and a 1.2-kb product of the nested PCR from infected W. somnifera samples. No PCR product was observed in the healthy counterparts. The PCR assay confirmed the presence of phytoplasma as causal agent. The PCR product was cloned with TOPO TA cloning kit (Invitrogen, Carlsbad, CA) and isolated plasmids were again assessed by restriction enzyme (EcoRI) digestion before sequencing. Purified plasmids were sequenced. Partially sequenced nucleotide sequence analysis of 16SrRNA gene cloned from W. somnifera phytoplasma showed high similarity with several isolates of the 16SrVI group of phytoplasmas. The highest nucleotide matching (99 and 98%) was observed with Centaurea solstitialis virescence phytoplasma (Genbank Accession No. AY270156) and Periwinkle little leaf phytoplasma (PPL-Bd; Genbank Accession No. AF 228053) reported in Italy and Bangladesh, respectively. In restriction fragment length polymorphism (RFLP) analysis, AluI, EcoRI, HhaI, HincII, KpnI, and Sau3AI (Promega, Madison, WI; 5 U per reaction) were used for comparison of restriction pattern of present/reference phytoplasma and with that previously reported (3). The present phytoplasma produced identical restriction profile to those of periwinkle infected by PPL-Bd (periwinkle little leaf phytoplasma, Bangladesh, group 16SrVI). On the basis of PCR studies, absence of virus particles under TEM in infected samples, RFLP analysis and nucleotide sequence matching with previously characterized phytoplasma, this phyto-plasma is classified as a member of Clover proliferation group (16SrVI). To our knowledge, this is the first report of a phytoplasma belonging to 16Sr VI group from W. somnifera. References: (1) S. Deng and C. Hiruki. J. Microbiol. Methods 14:53, 1991. (2) D. E. Gundersen and I.-M. Lee. Phytopathol. Mediterr.35:144, 1996. (3) I.-M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998. (4) M. Zaim and A. Samad. Plant Sci. 109:225,1995.
484. T. Samaddar et al., “Karyotype Analysis of Three Important Traditional Indian Medicinal Plants, Bacopa Monnieri, Tylophora Indica and Withania Somnifera,” The Nucleus, vol. 55, no. 1, pp. 17–20, Apr. 2012. doi: 10.1007/s13237-012-0048-2. Abstract
     Chromosome characters of three indigenous medicinal plants of high repute were studied. Their karyotypes were commonly mono-modal, decreasing in length from the longest to the shortest chromosomes. The chromosome complement in centromeric positions was 6sm+34M+24m for Bacopa monnieri (L.) Wettst. (2n\,= 64), 2m.st+12M+6sm+2m for Tylophora indica (Burm.f.) Merrill (2n\,= 22) and 2m.st+4m.sm+4M+18m+20sm for Withania somnifera (L.) Dunal (2n\,= 48).
485. Sanchita, S. Singh, and A. Sharma, “Bioinformatics Approaches for Structural and Functional Analysis of Proteins in Secondary Metabolism in Withania Somnifera,” Molecular Biology Reports, vol. 41, no. 11, pp. 7323–7330, Nov. 2014. doi: 10.1007/s11033-014-3618-3. Abstract
     Withania somnifera (Ashwagandha) is an affluent storehouse of large number of pharmacologically active secondary metabolites known as withanolides. These secondary metabolites are produced by withanolide biosynthetic pathway. Very less information is available on structural and functional aspects of enzymes involved in withanolides biosynthetic pathways of Withiana somnifera. We therefore performed a bioinformatics analysis to look at functional and structural properties of these important enzymes. The pathway enzymes taken for this study were 3-Hydroxy-3-methylglutaryl coenzyme A reductase, 1-Deoxy-d-xylulose-5-phosphate synthase, 1-Deoxy-d-xylulose-5-phosphate reductase, farnesyl pyrophosphate synthase, squalene synthase, squalene epoxidase, and cycloartenol synthase. The prediction of secondary structure was performed for basic structural information. Three-dimensional structures for these enzymes were predicted. The physico-chemical properties such as pI, AI, GRAVY and instability index were also studied. The current information will provide a platform to know the structural attributes responsible for the function of these protein until experimental structures become available.
486. Sanchita et al., “Physiological Performance, Secondary Metabolite and Expression Profiling of Genes Associated with Drought Tolerance in Withania Somnifera,” Protoplasma, vol. 252, no. 6, pp. 1439–1450, Nov. 2015. doi: 10.1007/s00709-015-0771-z. Abstract
     Physiological, biochemical, and gene expression responses under drought stress were studied in Withania somnifera. Photosynthesis rate, stomatal conductance, transpiration rate, relative water content, chlorophyll content, and quantum yield of photosystems I and II (PSI and PSII) decreased in response to drought stress. Comparative expression of genes involved in osmoregulation, detoxification, signal transduction, metabolism, and transcription factor was analyzed through quantitative RT–PCR. The genes encoding 1-pyrroline-5-carboxylate synthetase (P5CS), glutathione S-transferase (GST), superoxide dismutase (SOD), serine threonine-protein kinase (STK), serine threonine protein phosphatase (PSP), aldehyde dehydrogenase (AD), leucoanthocyanidin dioxygenase/anthocyanin synthase (LD/AS), HSP, MYB, and WRKY have shown upregulation in response to drought stress condition in leaf tissues. Enhanced detoxification and osmoregulation along with increased withanolides production were also observed under drought stress. The results of this study will be helpful in developing stress-tolerant and high secondary metabolite yielding genotypes.
487. O. Sangwan, R. Avtar, and A. Singh, “Genetic Variability, Character Association and Path Analysis in Ashwagandha [Withania Somnifera (L.) Dunal] under Rainfed Conditions.,” Research in Plant Biology, vol. 3, no. 2, pp. 32–36, 2013. https://www.cabdirect.org/cabdirect/abstract/20133182722. Abstract
     Twenty six diverse genotypes of ashwagandha [Withania somnifera (L.) Dunal] were evaluated for fourteen quantitative traits under rainfed conditions. Both phenotypic and genotypic coefficients of variation were higher for important traits like number of berries/plant, fresh weight of berries/plant, biomass yield at maturity and ratio of fresh above ground biomass: fresh root biomass. High...
488. R. S. Sangwan et al., “Withanolide A Biogeneration in in Vitro Shoot Cultures of Ashwagandha (Withania Somnifera DUNAL), a Main Medicinal Plant in Ayurveda,” Chemical and Pharmaceutical Bulletin, vol. 55, no. 9, pp. 1371–1375, 2007. doi: 10.1248/cpb.55.1371. Abstract
     Multiple shoot cultures of two experimental lines of Withania somnifera plants (RS-Selection-1 and RS-Selection-2) were established using nodal segments as explants. The hormonal combinations of benzyl adenine and kinetin not only influenced their morphogenetic response but also differentially modulated the level of biogeneration of withanolide A in the in vitro shoots of the two lines. Interestingly, withanolide-A, that was hardly detectable in the aerial parts of field-grown Withania somnifera (explant source), accumulated considerably in the in vitro shoot cultures of the plant. The productivity of withanolide A in the cultures varied considerably (ca. 10-fold, 0.014 to 0.14 mg per gram fresh weight) with the change in the hormone composition of the culture media as well as genotype used as source of the explant. The shoot culture of RS-Selection-1 raised at 1.00 ppm of BAP and 0.50 ppm of kinetin displayed the highest concentration of withanolide A in the green shoots of 0.238 g per 100 g dry weight tissue. This was a more analytical concentration keeping in view the isolation yields so far reported from the dried roots of the field-grown plant (ca. 0.015 g per 100 g dry weight), even if isolation losses are considered during purification. The enhanced de novo biogenesis of withanolide A in shoot cultures was corroborated with radiolabel incorporation studies using [2-14C] acetate as a precursor. Production of withaferin A was also found in the in vitro shoot cultures. As this compound is a predominant withanolide of native shoots as well and has been already reported to be accumulated in in vitro shoot cultures, its biogeneration observed in these shoot cultures is not discussed in detail.
489. R. S. Sangwan, N. D. Chaurasiya, P. Lal, L. Misra, R. Tuli, and N. S. Sangwan, “Withanolide A Is Inherently de Novo Biosynthesized in Roots of the Medicinal Plant Ashwagandha (Withania Somnifera),” Physiologia Plantarum, vol. 133, no. 2, pp. 278–287, 2008. doi: 10.1111/j.1399-3054.2008.01076.x. Abstract
     Ashwagandha (Withania somnifera Dunal., Solanaceae) is one of the most reputed medicinal plants of Ayurveda, the traditional medical system. Several of its traditionally proclaimed medicinal properties have been corroborated by recent molecular pharmacological investigations and have been shown to be associated with its specific secondary metabolites known as withanolides, the novel group of ergostane skeletal phytosteroids named after the plant. Withanolides are structurally distinct from tropane/nortropane alkaloids (usually found in Solanaceae plants) and are produced only by a few genera within Solanaceae. W. somnifera contains many structurally diverse withanolides in its leaves as well as roots. To date, there has been little biosynthetic or metabolism-related research on withanolides. It is thought that withanolides are synthesized in leaves and transported to roots like the tropane alkaloids, a group of bioactive secondary metabolites in Solanaceae members known to be synthesized in roots and transported to leaves for storage. To examine this, we have studied incorporation of 14C from [2-14C]-acetate and [U-14C]-glucose into withanolide A in the in vitro cultured normal roots as well as native/orphan roots of W. somnifera. Analysis of products by thin layer chromatography revealed that these primary metabolites were incorporated into withanolide A, demonstrating that root-contained withanolide A is de novo synthesized within roots from primary isoprenogenic precursors. Therefore, withanolides are synthesized in different parts of the plant (through operation of the complete metabolic pathway) rather than imported.
490. S. Saranya, R. Santhi*, K. Appavu, and K. Rajamani, “Soil Test Based Integrated Plant Nutrition System for Ashwagandha on Inceptisols,” Indian Journal of Agricultural Research, vol. 46, no. 1, pp. 88–90, 2012. https://arccjournals.com/journal/indian-journal-of-agricultural-research/.
491. A. Saravanakumar, A. Aslam, and A. Shajahan, “Development and Optimization of Hairy Root Culture Systems in Withania Somnifera (L.) Dunal for Withaferin-A Production,” African Journal of Biotechnology, vol. 11, no. 98, 2012. doi: 10.4314/ajb.v11i98. Abstract
     Transformation of Withania somnifera was carried out by using three Agrobacterium rhizogenes strains (ATCC 15834, R1000 and K599) for hairy root induction. Induction of hairy root was carried out in leaf, petiole and internodal explants. Hairy root induction was successful only in ATCC 15834 and R1000. The highest frequency of hairy root was obtained in petiole explants (64%) infected with R1000 and it resulted in five distinct morpho-types (callus (fragile), callus (hard), callus + hairy roots, hairy roots and callusing roots). The frequency of R1000 transformation was increased up to 93.2% by the addition of acetosyringone during various steps of infection. Molecular identification through PCR analysis of rolC confirmed the presence of Ri T-DNA. The half strength Murashige and Skoog (MS) liquid medium was found to be the best medium that supports the high root biomass accumulation than the other tested medium types (MS full strength, Gamborg B5 medium (B5) full strength and B5 half strength). High performance liquid chromatography (HPLC) analysis of hairy roots revealed the accumulation of withaferin-A (72.3 mg/g dw). This study reports the influence of Agrobacterium strains, explant types and acetosyringone in the hairy root induction of W. somnifera.Key words: Withania somnifera, hairy root, Agrobacterium rhizogenes, hairy root morpho-types.
492. G. Sardarkrushinagar, A. P. Balakrishnan, N. B. Patel, Dept. of Genetics and Plant Breeding, C. P. College of Agriculture, S. D. Agricultural University, Sardarkrushinagar, Gujarat (385 506), India, M. P. Patel, and Pulses Research Station, S. D. Agricultural University, Sardarkrushinagar, Gujarat (385 506), India, “Assessment of Heterosis for Root Yield and Attributing Traits in Ashwagandha [Withania Somnifera (L.) Dunal],” International Journal of Bio-resource and Stress Management, vol. 13, no. 1, pp. 93–98, Jan. 2022. doi: 20220209025844892. Abstract
     Astudy was conducted was conducted to estimate the extent of heterosis for root yield and attributing traits in ashwagandha at the Botanical Garden, Department of Genetics and Plant Breeding, C. P. C. A, S. D. Agricultural University, Sardarkrushinagar, Gujarat, India. Thirty six hybrids were generated from the diallel mating design of 9 diverse parents during October 2018 – March 2019 (rabi season). Fourty five entries were evaluated in RBD with 3 replications during October 2019–March 2020 (rabi season) for the yield and attribuiting traits. The root traits are given more emphasis because of its economic value.All the forty-five genotypes exhibited moderate to good mean performance along with an adequate amount of variability among and between the parents and hybrids. Most of the hybrids showed highly significant better parent and standard heterosis in desirable direction for the traits considered. The hybrids like SKA 10×JA 20, JA 134×AWS 1, SKA 24×SKA 26, SKA 24×AWS 1, SKA 11×AWS 1, SKA 11×SKA 26 and JA 20×AWS 1 were with a markedly significant heterotic response for fresh root yield trait. The results also suggest that the cross JA 134×AWS 1is appropriate to exploit heterosis in root length, fresh root yield and dry root yield. Further evaluation and generation advancement of these successful crosses can derive transgressive segregants and hybrids with high yield potential for commercialization.
493. K. V. Saritha and C. V. Naidu, “In Vitro Flowering of Withania Somnifera Dunal.—An Important Antitumor Medicinal Plant,” Plant Science, vol. 172, no. 4, pp. 847–851, Apr. 2007. doi: 10.1016/j.plantsci.2006.12.016. Abstract
     In vitro flowering, in vitro fruiting and effective micropropagation protocol were studied in Withania somnifera, an antitumor medicinal plant using axillary bud explants. The Murashige and Skoog’s medium (MS) supplemented with N6-benzyl adenine (BA) 2.0mgl−1 and α-naphthalene acetic acid 0.1mgl−1 was found optimum for production of multiple shoots. The regenerated plantlets were found to form tiny green floral buds after 4–6 weeks of culture in MS medium supplemented with Kinetin (0.5–4.0mgl−1) and indole-3-acetic acid (0.1mgl−1). In vitro fruiting was observed in the presence of Kn (2.0mgl−1) and IAA (0.1mgl−1). This paper describes in vitro flowering system to overcome problems associated with flower growth and development as well as fruit and seed production in vitro.
494. A. Saroj, A. Kumar, N. Qamar, M. Alam, H. N. Singh, and A. Khaliq, “First Report of Wet Rot of Withania Somnifera Caused by Choanephora Cucurbitarum in India,” Plant Disease, vol. 96, no. 2, pp. 293–293, Feb. 2012. doi: 10.1094/PDIS-09-11-0801. Abstract
     Withania somnifera (Ashwagandha) is native to India and commercially cultivated for the production of root withanolides that have anticarcinogenic properties. A disease appeared on plantings of W. somnifera during the 2010 monsoon at the CIMAP and in adjoining areas of northern India. Symptoms first appeared as water-soaked lesions on leaves and stems that progressed to a wet rot. Mature lesions harbored black fructifications of the suspect pathogen. Pathogen isolations were done by placing pieces of infected tissues on potato dextrose agar. A fungus tentatively identified as a Choanephora sp. that produced white aerial mycelia that later turned pale yellow was consistently isolated from infected plant parts. Mycelia were hyaline and nonseptate. Sporangiophores bearing sporangiola were erect, hyaline, unbranched, apically dilated to form a clavate vesicle from which arose dichotomously branched distally clavate secondary vesicles. Sporangiola were indehiscent, ellipsoid, brown to dark brown with distinct longitudinal striations, and measured 12 to 20 × 6 to 12 μm. Sporangia were multispored, spherical, initially white to yellow and pale brown to dark brown at maturity, and measured 40 to 160 μm. Sporangiospores from sporangia were ellipsoid to broadly ellipsoid, brown to dark brown, indistinctly striate with fine hyaline polar appendages, and measured 16 to 20 × 8 to 12 μm. On the basis of the cultural as well as morphological characteristics and description in the monograph by Kirk (2), the fungus was identified as a Choanephora sp. The identification was also confirmed by IMTECH, Chandigarh, India with Accession No. MTCC-10731. The species was later characterized as Choanephora cucurbitarum (Berk. & Ravenel) Thaxt (GenBank Accession No. AB470642) by using universal primers ITS-1 and ITS-4. Its sequence comprising of 18S rRNA partial, complete ITS 1, 5.8S rRNA, ITS 2, and 28S rRNA partial was submitted to NCBI GenBank with Accession No. JN639861. Pathogenicity of the fungus was established on five healthy plants by artificial inoculation with spray of an aqueous spore suspension containing 106 spores/ml. Plants sprayed with sterile distilled water were used as controls. Both inoculated and control plants were kept in a humidity chamber (96%) for 3 days and thereafter placed in the glasshouse at 28 ± 2°C. Initial symptoms developed in 2 to 3 days while typical disease symptoms appeared on all the inoculated plants after 7 to 10 days. Control plants were free from infection. The reisolation from artificially infected plants again yielded a Choanephora sp., thus fulfilling Koch’s postulates. W. somnifera cultivation has been affected by root rot and wilt caused by Fusarium solani and leaf spot caused by Alternaria dianthicola (3). The occurrence of a Choanephora sp. was reported on periwinkle, petunia (1), and Boerhavia diffusa (4). However, to our knowledge, incidence of this pathogen on W. somnifera has not been reported so far. Thus, wet rot of W. somnifera caused by C. cucurbitarum is a new report from India and worldwide. References: (1) G. E. Holcomb. Plant Dis. 87:751, 2003. (2) P. M. Kirk. Mycol. Pap. 152:1, 1984. (3) C. K. Maiti et al. Plant Dis. 91:467, 2007. (4) N. Singh et al. New Dis. Rep. 23:29, 2011.
495. A. Saroj et al., “New Report of Black Leaf Spot Mold (Pseudocercospora Fuligena) on Withania Somnifera from India,” Plant Disease, vol. 98, no. 9, pp. 1275–1275, Sep. 2014. doi: 10.1094/PDIS-03-14-0231-PDN. Abstract
     Withania somnifera (family solanaceae) commonly known as ashwagandha and Indian ginseng, originated in India is one of the most powerful medicinal plants for more than 3,000 years (1). It is commercially cultivated for its roots, a natural rich source of glycowithanolides, tannins, potassium nitrate, etc., which are an anti-inflammatory, anti-tumor, anti-oxidant, anti-ulcer, and regulator of the nervous system and sleep (2). During the monsoon of July 2011, black spots on the leaves of infected plants were observed in the ashwagandha growing Lucknow, Raibareilly, and adjoining areas of Uttar Pradesh province with 10 to 20% disease incidence. Early stage of disease were characterized by the presence of light chlorotic spots on both sides of old leaves that later turned into dark black spots resulting in early defoliation. About 27 samples were collected from different locations of the fields for isolation of the causal organism and microscopic studies. Infected leaves were cut into small pieces, surface sterilized with 1% sodium hypochlorite for 1 min, rinsed thrice with sterilized distilled water, and placed onto potato dextrose agar (PDA) plates. After 21 days of dark incubation at 25°C, 8- to 10-mm grayish-brown colonies were observed. Microscopic studies at early and mature stages of infection showed production of conidia in conidiophores. Conidiophores were mostly 5 to 9, few dense pale brown, simple unbranched, septate, geniculate and 14 to 55 × 3 to 5.5 μm. Conidia were subhyline, obclavate to cylindrical, some were straight to slightly curved, multiseptate, base long obconic to long obconically truncate, and 12 to 85 × 3.5 to 5 μm. On the basis of cultural and morphological studies, the pathogen was identified as Pseudocercospora fuligena (3). The pathogen identity was further confirmed at molecular level using universal primers ITS1/ITS4 through PCR (4). An amplification of the expected size (~550 bp) was generated, eluted from agarose gel by QIAquick gel extraction kit (Qiagen), cloned into pGEM-T Easy vector (Promega), sequenced, and deposited in GenBank (Accession No. KF881898). NCBI BLASTn showed 99% identity with P. fuligena (GU214675) strain CPC 12296, isolated from Lycopersicon sp. Pathogenicity test was carried out on 10 plants of W. somnifera cv. Poshita through two approaches, one using mycelia from culture and another using spore suspension from naturally infected leaves. In the first approach, fungal mycelia were applied onto the healthy ashwagandha leaves, whereas in the second approach, infected leaves were washed with distilled water and spore suspension of 106 spores/ml was sprayed on healthy plants. Plants sprayed with sterilized distilled water served as controls. Inoculated plants were placed in a growth chamber at 28°C under 90% humidity for 3 days. After, pots were placed in the glasshouse at 27 ± 2°C with 70 to 80% humidity for 21 days. Initial symptoms appeared on the 7th day while typical symptoms appeared on all the inoculated plants after 12 to 17 days. Control plants remained free of infection. Re-isolation of the pathogen on PDA fulfilled Koch’s postulates. Black leaf mold caused by P. fuligena has been reported on tomato (5). This is the first report of black leaf mould caused by P. fuligena on W. somnifera from India. P. fuligena has the potential to reduce yield of W. somnifera. References: (1) Anonymous. Alt. Med Rev. 9:211, 2004. (2) B. D. Basu and K. R. Kirtikar. Indian Medicinal Plants: Plates, vol. 1-4. Bishen Singh Mahendra Pal Singh, Dehradun, India, 1991. (3) T. C. Wang et al. Plant Dis. 79:661, 1995. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990. (5) S. Yamada. Ann. Phytopathol. Soc. Jpn. 15:13, 1951.
496. T. M. Sathees Kannan, S. Sownthariya, and S. Anbazhakan, “In Vitro Mass Propagation of Withania Somnifera Dunal Using Seaweed Extract,” International Letters of Natural Sciences, vol. 19, 2014. http://agro.icm.edu.pl/agro/element/bwmeta1.element.agro-f45ddf96-3b8b-45b5-808a-74d5753b1e27.
497. S. S. Saxena and I. D. Tyagi, “Effect of Gibber-Ellic Acid on the Growth of Hookah Tobacco (Nicotiana Rustica L.,” Labdev Journal of Science and Technology, B, vol. 8, no. 1, pp. 41–3, 1970. https://www.cabdirect.org/cabdirect/abstract/19711702583. Abstract
     Pre-sowing treatment of hookah tobacco seed with 10, 50 and 100 ppm. GA solutions for 1, 4 and 8 h markedly increased germination, plant height, stem diameter, number of leaves/plant, leaf size, and fresh and dry weight of a plant, compared with untreated seeds; treatment with 100 ppm. GA for 4 h gave the best results, followed by 50 ppm. GA for 8 h.-M.S.M.
498. P. Saxena, S. Ahlawat, A. Ali, S. Khan, and M. Z. Abdin, “Gene Expression Analysis of the Withanolide Biosynthetic Pathway in Hairy Root Cultures of Withania Somnifera Elicited with Methyl Jasmonate and the Fungus Piriformospora Indica,” Symbiosis, vol. 71, no. 2, pp. 143–154, Feb. 2017. doi: 10.1007/s13199-016-0416-9. Abstract
     Withania somnifera, also known as Indian ginseng is known to contain valuable bioactive compounds, called withanolides that structurally resemble ginsenosides of Panax ginseng. These compounds provide the basis of pharmacological relevance in traditional systems of medicine. In the present study, 150 hairy root lines of W. somnifera were induced of which nine fast growing lines were analysed for their growth and withanolide content. Hairy root line W9 was selected due to its high specific growth rate (0.196 ± 0.005 d−1) and high withanolide content. The response to different concentrations of elicitors (methyl jasmonate and P. indica cell homogenate) and various exposure durations was assessed in the W9 hairy root line. The withanolide content as well as the pattern of gene expression from MVA, MEP and sterol pathway, was evaluated using qPCR. Though gene expression and withanolide content were found to be elevated in almost all MeJ and CHP treatments, the exposure of hairy roots to 15 μM MeJ for 4 h gave the maximum withanolide yield. The results suggest that the elicitation potential of methyl jasmonate was higher than that of P. indica cell homogenate for increasing withanolide levels in hairy roots of W. somnifera.
499. P. Scartezzini, F. Antognoni, L. Conte, A. Maxia, A. Troìa, and F. Poli, “Genetic and Phytochemical Difference between Some Indian and Italian Plants of Withania Somnifera (L.) Dunal,” Natural Product Research, vol. 21, no. 10, pp. 923–932, Aug. 2007. doi: 10.1080/14786410701500169. Abstract
     The geographical distribution of Withania somnifera (L.) Dunal is quite wide. However, in Italy, this species is very rare and grows spontaneously only in Sicily and in Sardinia. The PCR–RAPD technique has been utilized in this work to determine the genetic relationship among Sicilian, Sardinian and Indian samples and the HPLC analysis of whitaferin A was used as a marker to evaluate the phytochemical differences. The genetic difference between Indian and Sicilian plants of W. somnifera turned out to be smaller than that between Indian and Sardinian plants of this species. The phytochemical analysis as well showed that the Sardinian specimen strongly differed from the Indian and Sicilian ones in its contents of withaferin A. Our results seem to confirm the hypothesis that the Italian populations of this species may not be indigenous but naturalised. Due to the high withaferin A content of the Sardinian samples, these plants could be used as a source for pharmaceutical purposes.
500. L. J. Schermeister, F. A. Crane, and R. F. Voigt, “Growth and Differentiation of Atropa Belladonna L. as Affected by Different Sources of Nitrogen,” Journal of the American Pharmaceutical Association, vol. 49, no. 11, pp. 694–697, 1960. doi: 10.1002/jps.3030491104. Abstract
     Growth and morphology were compared on plants furnished six levels of nitrate and three of ammonia in water culture. The production of dry weight with ammonia was significantly greater than with nitrate. Ratios of shoot/root and leaf/stem indicated that use of the nitrogen was very different at low vs. high levels, particularly with ammonia nitrogen.
501. F. Seehofer and H. W. Lorenz, “A Curing Chamber with Air Circulation / Klimakammer Mit Luftumwälzung,” Contributions to Tobacco & Nicotine Research, vol. 1, no. 2, pp. 35–38, Apr. 1961. doi: 10.2478/cttr-2013-0006. Abstract
     Abstract For chemical and physical tests samples of homogeneous tobaccos of uniform humidity are required. To this end a curing chamber was constructed allowing conditioning of cigarettes and tobacco in very short time. The new apparatus can also be used for determining the hygroscopic compensation of different blends. Compared with conventional machines operating according to the same principle with pressure reduction, saturated salt solutions and air circulation it offers considerable improvements regarding duration and uniformity of conditioning
502. J. Sen and A. K. Sharma, “Micropropagation of Withania Somnifera from Germinating Seeds and Shoot Tips,” Plant Cell, Tissue and Organ Culture, vol. 26, no. 2, pp. 71–73, Aug. 1991. doi: 10.1007/BF00036108. Abstract
     Shoot multiplication was achieved in vitro from shoot tips of aseptically germinated seedlings of Withania somnifera L. using low concentrations of 6-benzyladenine (BA), viz. 2.2, 4.4 and 8.9 μM. Maximum number of shoots were obtained when 2.3 μM 2,4-dichlorophenoxyacetic acid (2,4-D) or 2.5 μM indolebutyric acid (IBA) was added to medium containing 4.4 μM BA during initiation of shoot multiplication, but not when added later. Direct multiple shoot initiation was also obtained from germinating seeds in the presence of BA alone. Rooting was successful in excised shoots grown on growth regulator-free MS medium. Rooted shoots were successfully established in soil in a greenhouse.
503. E. Shafiyon and A. A. Ehsan Pour, “Effect of Vitamin E (Alpha-Tocopherol) on Some Physiological & Biochemical Parameters of Tobacco Plant (Nicotiana Rustica L.) under in Vitro Salt Stress,” Applied Biology, vol. 31, no. 4, pp. 81–99, Feb. 2019. doi: 10.22051/jab.2019.4226. Abstract
     Tocopherols, identified as vitamin E, are amphipathic molecules known as antioxidant & able to remove ROS in lipophilic conditions. In this study, tobacco plants were cultured on MS medium containing concentrations of 0, 100, 200 mM NaCl supplemented with 0, 100, 200 & 400 mg.L ˉ1 α-tocopherol for 4 weeks. Then growth parameters including fresh & dry weight, photosynthetic pigments including chlorophyll a, b, total chlorophyll & carotenoids, sodium & potassium, proline, total phenol, soluble sugar, anthocyanins, & total protein were measured. The results showed that, increasing of α-tocopherolconcentrations increased fresh & dry weight of plants & the amount of photosynthetic pigments, proline, soluble sugar, total phenol, anthocyanins & potassium while, amount of sodium was decreased significantly.Protein content increased by the highest concentration of α-tocopherol. It seemed that, treatment with vitamin E by changing physiological parameters increased salt tolerance of tobacco plant.
504. S. Shah, R. Saravanan, and N. A. Gajbhiye, “Phytochemical and Physiological Changes in Ashwagandha (Withania Somnifera Dunal) under Soil Moisture Stress,” Brazilian Journal of Plant Physiology, vol. 22, pp. 255–261, 2010. doi: 10.1590/S1677-04202010000400005. Abstract
     Ashwagandha, commonly known as the Indian Ginseng (Withania somnifera) is an ingredient in many herbal preparations used for healing various ailments. It is cultivated in semi-arid regions of India as rain-fed crop. The response of this crop (cv. JA-134) to progressive soil moisture deficit on growth, physiology and content of phytochemicals was studied. At the end of treatment period, soil water potential (Ψw) decreased to -10.93, -1.15 MPa at 30 cm depth, predawn leaf water potentials were reduced to -0.62 and -0.51 MPa in the severe and moderate stress. Leaf area was reduced by 27.4% and 34.4% in moderate and severe water stress treatment compared to control. Chlorophyll content was reduced by 60% in the severe water stress. Changes in the steroidal alkaloids and lactones that are the major medicinal active principles derived from root extracts of this plant species was also analyzed. A reduction of 52.6% in net photosynthesis was noticed under severe stress. At the end of the treatment period, moderate and severe stress treatments had 0.751 and 0.714 mg g-1 12-deoxywithastramonolide (12-DWS) content in the roots respectively. The content of 12-DWS and withanolide were at 77% and 93% of the control in the moderate and severe stress, respectively, while yield of these compounds were at 65% and 78% of control in mild and severe stress. Thus, preferential partitioning of biomass in roots leads to higher root yield in moderate stress. Higher root yield partially offset the loss of bioactive compounds under soil moisture stress.
505. B. P. S. Shahram and D. Z. Hamid, “Effects of Topping and Suckericide on Leaf Quality of Tobacco (Nicotiana Tabacum),” vol. 2, no. 3, pp. 723–731, Jan. 2014. https://www.sid.ir/en/journal/ViewPaper.aspx?id=377387. Abstract
     Download Free Full-Text of an article EFFECTS OF TOPPING AND SUCKERICIDE ON LEAF QUALITY OF TOBACCO (NICOTIANA TABACUM)
506. A. Shajahan et al., “An Efficient Hairy Root System for Withanolide Production in Withania Somnifera (L.) Dunal,” in Production of Plant Derived Natural Compounds through Hairy Root Culture, S. Malik, Ed. Cham: Springer International Publishing, 2017, pp. 133–143. doi: 10.1007/978-3-319-69769-7_7. Abstract
     Withania somnifera (L.) Dunal is one of the most important medicinal plant belonging to the family Solanaceae. Its root have been used as a drug since ancient times, and various pharmacological effects have been attributed to the occurrence of secondary compounds like withaferin-A and withanolide-A. Recently, huge interests are generated for production of these bioactive compounds through Agrobacterium rhizogenes-mediated hairy root culture techniques. The present review explores the culture conditions for efficient Agrobacterium-mediated hairy root culture system of W. somnifera for withanolide production. The hairy root induction is influenced by several factors like bacterial stain, type of explant, and cocultivation methods. The transformation efficiency could be enhanced by the addition of acetosyringone and SAAT treatments during cocultivation. Recent studies have also shown positive correlations of elicitors and biosynthetic pathway genes on withanolide production in hairy root culture of W. somnifera.
507. M. Sharada, A. Ahuja, and S. P. Vij, “Application of Biotechnology in Indian Ginseng (Ashwagandha): Progress and Prospects,” in Recent Advances in Plant Tissue Culture and Its Applications, New Delhi, India: I.K. International Publishers Pvt. Ltd., 2008, p. 694. https://www.researchgate.net/publication/231228099_Application_of_Biotechnology_in_Indian_Ginseng_Ashwagandha_Progress_and_prospects. Abstract
     Medicinal plant biotechnology offers many uncommon opportunities and techniques to conserve, propagate, improve and utilize medicinal plants and herbs in a better fashion. Ever since the idea of cellular totipotency, contemporary developments in the area of plant tissue culture for devising methods for rapid and round the year multiplication of desired genotypes, production of pathogen-free stocks, raising uniform clones from highly heterozygous plants, monitoring production of useful natural products in vitro, propagating plants with changed/altered genotypes including the ones genetically transformed have added new dimensions and pushed this science to realm of technological applications by providing the base for modern biotechnology. In India, cultivation of medicinal plants is limited to some selected species including Indian Ginseng (Withania somnifera) which synthesizes withanolides and/or related compounds but its wide spread chemodiversity is not conducive for their commercial isolation from natural populations. Plant tissue culture offers opportunity for improvement of Ashwagandha in many ways such as, development of a mass propagation system, to obtain somaclones or mutants with desired chemical constituents, genetic transformations and in vitro production of biologically active metabolites. The focus of the present review is the application of tissue culture technology, progress made and further scope on the application of biotechnology in this commercially important promising Indian medicinal plant. Also, we describe the results of in vitro studies on regeneration and production of selected biologically active withanolides obtained in our laboratory.
508. M. Sharada et al., “Withanolide Production by in Vitro Cultures of Withania Somnifera and Its Association with Differentiation,” Biologia Plantarum, vol. 51, no. 1, pp. 161–164, Mar. 2007. doi: 10.1007/s10535-007-0031-y. Abstract
     Withanolides-steroidal lactones, isolated from various Solanaceous plants have received considerable attention due to their potential biological activities. Five selected withanolides (withanone, withaferin A, withanolide A, withanolide B, withanolide E) were identified by HPLC-UV (DAD) — positive ion electrospray ionization mass spectroscopy in Withania somnifera (L.) Dunal cv. WSR plants and tissues cultured in vitro at different developmental phases. Cultures were established from five explants on Murashige and Skoog’s medium supplemented with different plant growth regulators. Results suggest that production of withanolides is closely associated with morphological differentiation.
509. R. Sharma, H. Singh, M. Kaushik, R. Nautiyal, and O. Singh, “Adaptive Physiological Response, Carbon Partitioning, and Biomass Production of Withania Somnifera (L.) Dunal Grown under Elevated CO2 Regimes,” 3 Biotech, vol. 8, no. 6, p. 267, May 2018. doi: 10.1007/s13205-018-1292-1. Abstract
     Winter cherry or Ashwagandha (Withania somnifera) is an important medicinal plant used in traditional and herbal medicine system. Yet, there is no information available on response of this plant to changing climatic conditions particularly elevated atmospheric CO2 concentrations. Therefore, we conducted an experiment to examine the effect of elevated CO2 concentrations (ECs) on Withania somnifera. The variations in traits of physiological adaptation, net primary productivity, carbon partitioning, morphology, and biomass in response to elevated CO2 concentrations (ambient, 600 and 800 µmol mol−1) during one growth cycle were investigated within the open top chamber (OTC) facility in the foothill of the Himalayas, Dehardun, India. ECs significantly increased photosynthetic rate, transpiration rate, stomatal conductance, water use efficiency, soil respiration, net primary productivity and the carbon content of plant tissues (leaf, stem, and root), and soil carbon. Furthermore, ECs significantly enhanced biomass production (root and shoot), although declined night leaf respiration. Overall, it was summarized that photosynthesis, stomatal conductance, water use efficiency, leaf, and soil carbon and biomass increased under ECs rendering the physiological adaptation to the plant. Increased net primary productivity might facilitate mitigation effects by sequestering elevated levels of carbon dioxide. We advocate further studies to investigate the effects of ECs on the accumulation of secondary metabolites and health-promoting substances of this as well as other medicinal plants.
510. P. Sharma and R. Pandey, “Biological Control of Root-Knot Nematode; Meloidogyne Incognita in the Medicinal Plant; Withania Somnifera and the Effect of Biocontrol Agents on Plant Growth,” African Journal of Agricultural Research, vol. 4, no. 6, pp. 564–567, Jun. 2009. doi: 10.5897/AJAR.9000087. Abstract
     The medicinally important plant; Withania somnifera, is highly susceptible to root knot nematode; Meloidogyne incognita. Various nematode antagonistic fungi have been studied for their use as biocontrol agents. In the experiment, the potential of fungi Trichoderma harzianum, Paecilomyces lilacinus and Arthrobotrys oligospora along with natural organic compound (Neem compound mix) to control the nematode; M. incognita was evaluated. Also, their potential to control nematodes was compared with that achieved by using the chemical control agent; carbofuran. The fungal agents evaluated significantly controlled nematode population and enhanced plant growth.
511. A. Sharma, S. Rana, G. A. Rather, P. Misra, M. K. Dhar, and S. K. Lattoo, “Characterization and Overexpression of Sterol Δ22-Desaturase, a Key Enzyme Modulates the Biosyntheses of Stigmasterol and Withanolides in Withania Somnifera (L.) Dunal,” Plant Science, vol. 301, p. 110642, Dec. 2020. doi: 10.1016/j.plantsci.2020.110642. Abstract
     Withanolides constitute an extensive and vital class of metabolites displaying wide array of structural and therapeutic properties with unique side-chain modifications. These show diversified scaffolds and are promising pharmaceutical molecules with well documented anti-inflammatory and anti-cancer properties. Sterols are dynamic class of compounds and essential molecules having structural and functional significance. These contribute to the synthesis of withanolides by providing structural precursors. In this context, we have characterized sterol Δ22-desaturase from Withania somnifera and also functionally validating it by confirming its desaturase nature in conjunction with quantitative real-time expression profiling and metabolite evaluation. Further, transgenic hairy roots of W. somnifera displayed a higher accumulation of stigmasterol and withanolides. The increase in chemical constituents was concomitant with an increased gene copy number predicted via Southern blotting. Additionally, transgenic lines of tobacco over-expressing WsCYP710A11 displayed a substantial increase in its expression, corroborating well with enhanced stigmasterol content. Characterization of CYP710A11 from W. somnifera and its homologous transgenic expression has demonstrated its role in the regulation of withanolides biosynthesis. It also exhibited a differential transcriptional profile in response to exogenous elicitations. These empirical findings suggest the crucial role of CYP710A11 in stigmasterol biosynthesis. This in turn has implications for the overproduction of withanolides via pathway channelling.
512. A. Sharma and P. K. Pati, “First Report of Withania Somnifera (L.) Dunal, as a New Host of Cowbug (Oxyrachis Tarandus, Fab.) In Plains of Punjab, Northern India,” p. 3, 2011. Abstract
     During February-June 2009-10, cowbugs were found on aerial apical parts of Ashwagandha (Withania somnifera) plants in the Amritsar District of Punjab. The cowbugs fed on apical portions of the stem, making them rough and woody in appearance, brown in colour that gradually dried and apical leaves were shed off. These cowbugs were identified as Oxyrachis tarandus [1] (Hemiptera: Membracidae). To the best of our knowledge, this is the first report of Withania somnifera (L.) Dunal as a new host for O. tarandus in Punjab province of Northern India.
513. A. Sharma and P. Kumar Pati, “First Record of the Carmine Spider Mite, Tetranychus Urticae , Infesting Withania Somnifera in India,” Journal of Insect Science, vol. 12, no. 50, pp. 1–4, Apr. 2012. doi: 10.1673/031.012.5001.
514. A. Sharma, G. A. Rather, P. Misra, M. K. Dhar, and S. K. Lattoo, “Gene Silencing and Over-Expression Studies in Concurrence With Promoter Specific Elicitations Reveal the Central Role of WsCYP85A69 in Biosynthesis of Triterpenoids in Withania Somnifera (L.) Dunal,” Frontiers in Plant Science, vol. 10, 2019. doi: 10.3389/fpls.2019.00842. Abstract
     Withania somnifera (Ashwagandha) synthesizes a wide spectrum of triterpenoids that are produced via an intricate isoprenoid pathway whose biosynthetic and regulatory mechanism remains elusive. Their pharmacological examination position them as potent bioactive molecules, hence demanding their copious production. Previous investigations have revealed that P450 monooxygenases are pivotal enzymes involved in the biosynthetic machinery of various metabolites and assist in decorating their core skeletal structures. The present study entails the isolation and functional characterization of castasterone synthase (CYP85A69) from W. somnifera. The full length WsCYP85A69, having an open reading frame of 1413 bp, encodes 470 amino acid residues. Further, in vitro conversion of 6-deoxocastasterone into castasterone validated its oxidative functionality. Product formation was confirmed using LC-PDA-MS with a m/z value of 506 [M+ACN]+. In planta transient over-expression of WsCYP85A69 significantly enhanced castasterone, stigmasterol and withanolides (WS-I, WS-II, WS-III). Artificial micro-RNA mediated silencing of WsCYP85A69 resulted in the reduced accumulation of castasterone, stigmasterol and withanolides (WS-I, WS-II, WS-III). Altogether, these non-complementary approaches plausibly suggest a key role of WsCYP85A69 in the biosynthesis of castasterone and the accumulation of withanolides and stigmasterol. Furthermore, a promoter analysis of WsCYP85A69 resulted in the identification of several potential cis-regulatory elements. Elicitations, given on the basis of identified cis-regulatory elements, demonstrated methyl jasmonate as an effective inducer of WsCYP85A69. Overall, these empirical findings suggest that functional characterization of WsCYP85A69 may conceivably be helpful to unravel the mechanism of brassinosteroids biosynthesis and could also pave the way for targeted metabolic engineering.
515. A. Sharma, G. A. Rather, P. Misra, M. K. Dhar, and S. K. Lattoo, “Jasmonate Responsive Transcription Factor WsMYC2 Regulates the Biosynthesis of Triterpenoid Withanolides and Phytosterol via Key Pathway Genes in Withania Somnifera (L.) Dunal,” Plant Molecular Biology, vol. 100, no. 4, pp. 543–560, Jul. 2019. doi: 10.1007/s11103-019-00880-4. Abstract
     Functional characterization of WsMYC2 via artificial microRNA mediated silencing and transient over-expression displayed significant regulatory role vis-à-vis withanolides and stigmasterol biosyntheses in Withania somnifera. Further, metabolic intensification corroborated well with higher expression levels of putative pathway genes. Additionally, copious expression of WsMYC2 in response to exogenous elicitors resulted in enhanced withanolides production.
516. A. Sharma, I. Sharma, and P. K. Pati, “Post-Infectional Changes Associated with the Progression of Leaf Spot Disease in Withania Somnifera,” Journal of Plant Pathology, vol. 93, no. 2, pp. 397–405, 2011. https://www.jstor.org/stable/41999011. Abstract
     Leaf spot caused by Alternaria alternata is a prevalent disease of Withania somnifera, a high value medicinal plant. The severity of infection is closely related with the progression of the disease. In the present work, various biochemical parameters such as free proline, total chlorophyll, total protein, lipid peroxidation, ion leakage and the activity of some antioxidant enzymes were analyzed to assess the post infectional changes associated with infection. As disease progressed, a decrease of chlorophyll and total protein content was observed. The lowest level of total chlorophyll (16 fold decrease) was observed when the severity of infection was at its peak. A significant increase in free proline content, lipid peroxidation and ion leakage was also observed. Antioxidant enzymes behaved in a differential way. Catalase, ascorbate peroxidase, guaiacol peroxidase and superoxide dismutase specific activities showed a significant increase concomitantly with disease progression. By contrast, glutathione reductase and monodehydroascorbate reductase decreased significantly (2.5 and 6.5 folds, respectively) as disease progressed. Besides Superoxide dismutase, the other antioxidant enzymes analysed did not have a significant correlation with the number of spots/cm² leaf area. Although the present study suggests a complex pattern of post infectional changes, the information obtained is useful for a better understanding of host pathogen interaction.
517. V. Sharma, A. P. Gupta, P. Bhandari, R. C. Gupta, and B. Singh, “A Validated and Densitometric HPTLC Method for the Quantification of Withaferin-A and Withanolide-A in Different Plant Parts of Two Morphotypes of Withania Somnifera,” Chromatographia, vol. 66, no. 9, pp. 801–804, Nov. 2007. doi: 10.1365/s10337-007-0396-2. Abstract
     A simple, sensitive and accurate high performance thin layer chromatographic (HPTLC) method has been developed for the estimation of withaferin-A and withanolide-A in different plant parts such as, leaf, root, stem and fruit of two morphotypes of Withania somnifera. HPTLC of W. somnifera methanolic extract was performed on Si 60 F254 (20 cm × 20 cm) plates with toluene:ethyl acetate:formic acid (5:5:1), as mobile phase. Quantitative evaluation of the plate was performed in the absorption-reflection mode at 530 nm. The method was validated for precision, repeatability, and accuracy. The average recovery of withaferin-A and withanolide-A in two levels were 96.0 and 96.7%, showing the excellent reproducibility of the method. The calibration curves were linear for both in the range of 200–3,200 ng. The technique has been applied, for the first time, for the estimation of withaferin-A and withanolide-A in different parts of the two morphotypes of Withania somnifera. The method is simple, precise, specific, sensitive and accurate and can be used for routine analysis as well as for quality control of raw materials and herbal formulations.
518. Q. Shi, C. Li, and F. Zhang, “Nicotine Synthesis in Nicotiana Tabacum L. Induced by Mechanical Wounding Is Regulated by Auxin,” Journal of Experimental Botany, vol. 57, no. 11, pp. 2899–2907, Aug. 2006. doi: 10.1093/jxb/erl051. Abstract
     The effects of different kinds of mechanical wounding on nicotine production in tobacco plants were compared, with sand or hydroponics culture under controlled conditions. Both removal of the shoot apex and damage of the youngest unfolded leaves nos 1 and 2 by a comb-like brusher with 720 punctures caused an increase in nicotine concentration in whole plants at day 3, and reached its highest level at day 6. The nicotine concentration induced by excision of the shoot apex was much higher than that induced by leaf wounding. Both treatments also caused an increase in jasmonic acid (JA) concentration within 90 min in the shoot, followed by an increase in the roots (210 min), in which the JA concentration induced by leaf wounding was significantly higher than that induced by excision of the shoot apex. The increase in nicotine concentration occurred throughout the whole plant, especially in the shoot, while the increase in JA concentration in the shoot was restricted to the damaged tissues, and was not observed in the adjacent tissues. Removal of the lateral buds that emerged after excision of the shoot apex caused a further increase in nicotine concentrations in the plant tissues. Removal of mature leaves, however, did not cause any changes in nicotine concentration in the plant, even though the degree of wounding in this case was comparable with that occurring with apex removal. The results suggest that the nicotine production in tobacco plants was not correlated with the degree of wounding (cut-surface or punctures), but was highly dependent on the removal of apical meristems and hence on the major sources of auxin in the plant. Furthermore, immediate application of 1-naphthylacetic acid (NAA) on the cut surface after removing the shoot apex completely inhibited the increase both in nicotine in whole plants and in JA in the damaged stem segment and roots. Application of an auxin transport inhibitor around the stem directly under the shoot apex of intact plants also caused an increase in nicotine concentration in the whole plant. The results strongly suggest that auxin serves as a negative signal to regulate nicotine synthesis in roots of tobacco plants.
519. H. B. Shilpashree, S. J. Sudharshan, A. K. Shasany, and D. A. Nagegowda, “Molecular Characterization of Three CYP450 Genes Reveals Their Role in Withanolides Formation and Defense in Withania Somnifera, the Indian Ginseng,” Scientific Reports, vol. 12, no. 1, p. 1602, Jan. 2022. doi: 10.1038/s41598-022-05634-9. Abstract
     The medicinal properties of Ashwagandha (Withania somnifera) are attributed to triterpenoid steroidal lactones, withanolides, which are proposed to be derived from phytosterol pathway, through the action of cytochrome P450 (CYP450) enzymes. Here, we report the characterization of three transcriptome-mined CYP450 genes (WsCYP749B1, WsCYP76 and WsCYP71B10), which exhibited induced expression in response to methyl jasmonate treatment indicating their role in secondary metabolism. All three WsCYP450s had the highest expression in leaf compared to other tissues. In planta characterization of WsCYP450s through virus induced gene silencing (VIGS) and transient overexpression approaches and subsequent metabolite analysis indicated differential modulation in the accumulation of certain withanolides in W. somnifera leaves. While WsCYP749B1-vigs significantly enhanced withaferin A (~ 450%) and reduced withanolide A (~ 50%), its overexpression drastically led to enhanced withanolide A (> 250%) and withanolide B (> 200%) levels and reduced 12-deoxywithastramonolide (~ 60%). Whereas WsCYP76-vigs led to reduced withanolide A (~ 60%) and its overexpression increased withanolide A (~ 150%) and reduced 12-deoxywithastramonolide (~ 60%). Silencing and overexpression of WsCYP71B10 resulted in significant reduction of withanolide B (~ 50%) and withanolide A (~ 60%), respectively. Further, while VIGS of WsCYP450s negatively affected the expression of pathogenesis-related (PR) genes and compromised tolerance to bacteria P. syringae DC3000, their overexpression in W. somnifera and transgenic tobacco led to improved tolerance to the bacteria. Overall, these results showed that the identified WsCYP450s have a role in one or several steps of withanolides biosynthetic pathway and are involved in conferring tolerance to biotic stress.
520. A. Shinde, P. Gahunge, and S. Rath, “Conservation and Sustainability of Ashwagandha: A Medicinal Plant,” Journal of Biological & Scientific Opinion, vol. 3, pp. 94–99, Apr. 2015. doi: 10.7897/2321-6328.03220. Abstract
     Ayurveda become a globally popular healthcare sy stem leading to constant rise in the demand for herbal medicines resulting in increased world herbal trade which stands at US 120 billion and is expected to reach US 7 trillion by 2050. As majority of Ayurvedic medicines are plant based, this demand has resulted in a huge pressure on the traditional sources of raw materials. Obviously, the wild sources are not able to meet the demand of raw materials. To overcome this situation alternate sources of raw materials like cultivated fields are look for supply. Dwindling forest cover, unscientific and exploitative collection leads to loss of supply source and threat to germplasm. Germplasm conservation and cultivation seems to be the best way forward for sustainable utilization of medicinal plants. Ashwagandha ( Withania somnifera Dunal) is widely used, prioritised Ayurvedic herb having annual demand 7000 tonnes /yr but its actual production is 1500 tonnes/yr. It grows naturally in subtropical region and is collected from wild and fields. Its r ampant collection has once leaded to critical pressure situation on its germplasm. This was noticed and a lot of effort was put in for its sustainable production. There i s lot of evidence we will discuss in the various research field of Ashwagandha like Agronomic research , Phy tochemical and Clinical research and Market Linkage . Ashwagndha due to such sustained and concerted multi - disciplinary effort has come near to a sustainable use mode. It is thus logical to extend the Ashwagandha model to other high demand medicinal plants and its feasibility.
521. N. Shitan, M. Morita, and K. Yazaki, “Identification of a Nicotine Transporter in Leaf Vacuoles of Nicotiana Tabacum,” Plant Signaling & Behavior, vol. 4, no. 6, pp. 530–532, Jun. 2009. doi: 10.4161/psb.4.6.8588. Abstract
     Alkaloids are nitrogen-containing low-molecular-weight organic substances that have a wide variety of chemical structures and often show diverse biological activities. Some alkaloids are transported from the source organ after their biosynthesis and moved to sink organs via long distance transport. One representative of such translocation of alkaloids is nicotine, a pyridine alkaloid, which is biosynthesized in root tissues, then translocated to the leaves, and finally accumulated in the leaf vacuoles in Nicotiana species. Although it is more than 10 years ago since nicotine translocation was identified, no transport protein has been characterized concerning the inter-organ movement of this alkaloid.
522. S. Shrivastava and P. K. Dubey, “In-Vitro Callus Induction and Shoot Regeneration in Withania Somnifera (L.) Dunal,” International Journal of Biotechnology & Biochemistry, vol. 3, no. 1, pp. 1–13, Jan. 2007. https://go.gale.com/ps/i.do?p=AONE&sw=w&issn=09732691&v=2.1&it=r&id=GALE%7CA172131882&sid=googleScholar&linkaccess=abs. Abstract
     Gale Academic OneFile includes In-vitro callus induction and shoot regeneration in Wit by S. Shrivastava and P.K. Dubey. Click to explore.
523. A. K. Shrivastava and P. K. Sahu, “Economics of Yield and Production of Alkaloid of Withania Somnifera (L.) Dunal,” American Journal of Plant Sciences, vol. 04, no. 10, p. 2023, Sep. 2013. doi: 10.4236/ajps.2013.410253. Abstract
     The yield parameters and cost economics of Withania somnifera were studied using Integrated Nutrient Management (INM) treatments. Withania somnifera is cultivated in around 10,780 ha with a production of 8429 tones in India. The annual demand of this herb increased from 7028 tones (2001-2002) to 9127 tones (2004-2005). The field experiment was conducted at the research farm of Department of Plant Physiology, College of Agriculture, Jabalpur during Kharif season of 2008-2009 (1st year) and 2009-2010 (2nd year), whereas the laboratory work was carried out in Quality Laboratory, College of Agriculture, Jabalpur. The soil of experimental field was sandy loam in texture with pH 7.5, EC 0.18 dsm-1 having good drainage. Soil analysis revealed that available nitrogen was low (202.0 kg·ha-1) whereas available phosphorus (16.25 kg·ha-1) and potassium (236.0 kg·ha-1) were in the medium range. The present paper shows how to determine the economics of varying INM treatments. Cultivation of W. somnifera in India is gaining popularity among farmers; however, due to poor soil fertility, and costly chemical fertilizers and pesticides its production is not economical or profitable.
524. A. Shrivastava, V. Upadhyay, D. Gautam, S. Sarvade, and R. Sahu, “Effect of Integrated Nutrient Management on Growth and Productivity of Withania Somnifera (L.) Dunal in Kymore Plateau and Satpura Hills of Madhya Pradesh, India,” Archives of Agriculture and Environmental Science, vol. 3, pp. 202–208, Jun. 2018. doi: 10.26832/24566632.2018.0302015. Abstract
     The field experiment was carried out at the Dusty Acres Farm, Jawaharlal Nehru Krishi Vishwa Vidyalaya (JNKVV), Jabalpur, Madhya Pradesh to study the effect of integrated nutrient man�agement for growth and yield of Withania somnifera (L.) Dunal (Variety: Jawahar Ashwagandha- 134). Tallest plants (49.35 cm) were observed at harvest stage, whereas higher number of branches per plant (5.78) of W. somnifera was observed at 90 DAS with T11, respectively. The mean number of leaves per plant (125.40) and LAI (10.345) of W. somnifera was higher at 90 DAS with T11. Mean CGR was maximum (2.536 g m-2 week-1 ) and mean RGR was highest (0.098 gg-1 day-1 ) of W. somnifera at 90 DAS and thereafter decline at 120 DAS and harvest stage with T11. Whereas, mean dry matter (1392.60 kg/ha) production of W. somnifera was higher at harvesting stage of crop with T11. Significantly higher mean root length (16.30 cm), root girth (2.26 cm) and mean dry root yield (612.8 kg/ha) of W. somnifera was recorded under T11 than the rest of treatments. Mean number of berries per plant (30.78) and mean number of seeds per berry (194.17) of W. somnifera were significantly higher under T11. Higher mean seed yield (62.6 kg/ha) and harvest index (43.61%) of W. somnifera was observed under T11 as compared to other treatments. Therefore, 100% recommended dose of NPK through fertiliz�ers along with vermicompost (2.5 t/ha), FYM (5.0 t/ha) and ZnSO4 20kg/ha (T11) followed by 50% recommended dose of NPK through fertilizers along with vermicompost (2.5 t/ha), FYM (5.0 t/ha) and ZnSO4 20kg/ha (T14) and 100% recommended dose of NPK through fertilizers along with vermicompost (2.5 t/ha) and ZnSO4 20kg/ha (T10) were found to be better integrated nutrient management for the cultivation of W. somnifera. �
525. D. D. Shukla, N. Bhattarai, and B. Pant, “In-Vitro Mass Propagation of Withania Somnifera (L.) Dunal,” Nepal Journal of Science and Technology, vol. 11, pp. 101–106, 2010. doi: 10.3126/njst.v11i0.4131. Abstract
     Ashwagandha (Withania somnifera L.) Dunal] is an important medicinal plant and a major source of alkaloids and steroids (withanolids), which is regularly used in pharmaceutical industries. Various vegetative parts were studied for its mass propagation through tissue culture technique. Seeds were pretreated with GA3 (50 and 100 mgl-1) for 24 h and 80% germination was achieved. All the explants were taken from in-vitro germinated plant. Among the different explants tested, multiple shoot formation was achieved from shoot-tip and nodal explants in MS medium + 0.25, 0.5, and 1.0 mgl-1 kinetin. Nodal explants were selected for mass propagation protocol because it formed maximum number of shoots (16.25 shoots per explant) on MS medium + 1mgl-1 kinetin after eight weeks of culture. Increase in concentration of kinetin was most effective for callus formation. For further multiplication these shoots were sub-cultured on MS +0.5 mgl-1 kinetin. Presence of IAA at 0.5 mgl-1 was most effective medium for rooting of in-vitro propagated shoots. However, hardening was not achieved for these propagated plants. Key words: IAA; IBA; NAA; kinetin; in-vitro multiplication DOI: 10.3126/njst.v11i0.4131Nepal Journal of Science and Technology 11 (2010) 101-106
526. O. P. Sidhu, S. Annarao, S. Chatterjee, R. Tuli, R. Roy, and C. L. Khetrapal, “Metabolic Alterations of Withania Somnifera (L.) Dunal Fruits at Different Developmental Stages by NMR Spectroscopy,” Phytochemical Analysis, vol. 22, no. 6, pp. 492–502, 2011. doi: 10.1002/pca.1307. Abstract
     Introduction Withania somnifera (Ashwagandha) is a high-value Ayurvedic medicinal plant and an important constituent of several dietary supplements. In order to substantiate the health claims, the herb has drawn considerable scientific attention. Objective The objective of the study was to investigate the alterations in primary and secondary metabolites of W. somnifera fruits during its maturity using NMR spectroscopy. Methodology Fruits at different stages of development from one week after fertilisation until maturity, classified in seven developmental stages, were analysed by a combined use of one- and two-dimensional NMR experiments. Results Seventeen metabolites were characterised and quantified from non-polar and polar extracts of different fruit development stages of W. somnifera. The principal component analysis of polar metabolites at different stages could be grossly classified into three metabolic phases, viz. initial phase, developmental phase and maturation phase. Conclusion Qualitative and quantitative analysis of metabolites in W. somnifera fruits indicated specific stages when fruits can be harvested for obtaining substantial bioactive ingredients for desirable pharmacological activity. This study potentially provides a complementary tool for quality control of herbal medicinal products when W. somnifera fruits are used. Copyright © 2011 John Wiley & Sons, Ltd.
527. N. Sierro et al., “The Impact of Genome Evolution on the Allotetraploid Nicotiana Rustica – an Intriguing Story of Enhanced Alkaloid Production,” BMC Genomics, vol. 19, p. 855, Nov. 2018. doi: 10.1186/s12864-018-5241-5. Abstract
     Background Nicotiana rustica (Aztec tobacco), like common tobacco (Nicotiana tabacum), is an allotetraploid formed through a recent hybridization event; however, it originated from completely different progenitor species. Here, we report the comparative genome analysis of wild type N. rustica (5 Gb; 2n\,= 4x\,= 48) with its three putative diploid progenitors (2.3–3 Gb; 2n\,= 2x =24), Nicotiana undulata, Nicotiana paniculata and Nicotiana knightiana. Results In total, 41% of N. rustica genome originated from the paternal donor (N. undulata), while 59% originated from the maternal donor (N. paniculata/N. knightiana). Chloroplast genome and gene analyses indicated that N. knightiana is more closely related to N. rustica than N. paniculata. Gene clustering revealed 14,623 ortholog groups common to other Nicotiana species and 207 unique to N. rustica. Genome sequence analysis indicated that N. knightiana is more closely related to N. rustica than N. paniculata, and that the higher nicotine content of N. rustica leaves is the result of the progenitor genomes combination and of a more active transport of nicotine to the shoot. Conclusions The availability of four new Nicotiana genome sequences provide insights into how speciation impacts plant metabolism, and in particular alkaloid transport and accumulation, and will contribute to better understanding the evolution of Nicotiana species. Electronic supplementary material The online version of this article (10.1186/s12864-018-5241-5) contains supplementary material, which is available to authorized users.
528. B. Sil, C. Mukherjee, S. Jha, and A. Mitra, “Metabolic Shift from Withasteroid Formation to Phenylpropanoid Accumulation in Cryptogein-Cotransformed Hairy Roots of Withania Somnifera (L.) Dunal,” Protoplasma, vol. 252, no. 4, pp. 1097–1110, Jul. 2015. doi: 10.1007/s00709-014-0743-8. Abstract
     Cotransformed hairy roots containing a gene that encodes a fungal elicitor protein, β-cryptogein, were established in Withania somnifera, a medicinal plant widely used in Indian systems of medicine. To find out whether β-cryptogein protein endogenously elicits the pathway of withasteroid biosynthesis, withaferin A and withanolide A contents along with transcript accumulation of farnesyl pyrophosphate (FPP) synthase, 3-hydroxy-3-methyl-glutaryl-CoA reductase (HMGR), and sterol glycosyltransferase (SGT) were analyzed in both cryptogein-cotransformed and normal hairy roots of W. somnifera. It was observed that the withaferin A and withanolide A contents were drastically higher in normal hairy roots than cryptogein-cotransformed ones. Similar trends were also observed on the levels of transcript accumulation. Subsequently, the enzyme activity of phenylalanine ammonia lyase (PAL), one of the key enzymes of phenylpropanoid pathway, was measured in both cryptogein-cotransformed and normal hairy roots of W. somnifera along with the levels of PAL transcript accumulation. Upliftment of PAL activity was observed in cryptogein-cotransformed hairy roots as compared to the normal ones, and the PAL expression also reflected a similar trend, i.e., enhanced expression in the cryptogein-cotransformed lines. Upliftment of wall-bound ferulic acid accumulation was also observed in the cryptogein-cotransformed lines, as compared to normal hairy root lines. Thus, the outcome of the above studies suggests a metabolic shift from withanolide accumulation to phenylpropanoid biosynthesis in cryptogein-cotransformed hairy roots of W. somnifera.
529. S. J. Sinclair, R. Johnson, J. D. Hamill, S. J. Sinclair, R. Johnson, and J. D. Hamill, “Analysis of Wound-Induced Gene Expression in Nicotiana Species with Contrasting Alkaloid Profiles,” Functional Plant Biology, vol. 31, no. 7, pp. 721–729, Jul. 2004. doi: 10.1071/FP03242. Abstract
     We determined the capacity of three Nicotiana (Solanaceae) species with very different alkaloid profiles (Nicotiana sylvestris Speg & Comes, Nicotiana alata Link & Otto and Nicotiana glauca Grah.) to increase their alkaloid contents in both leaf and root tissues following foliage damage. We also investigated the transcriptional responses of genes encoding enzymes important for alkaloid biosynthesis, namely quinolinate phosphoribosyltransferase (QPT), putrescine N-methyltransferase (PMT), ornithine decarboxylase (ODC) and the putative alkaloid biosynthetic gene A622. In response to wounding of foliage in the well studied ‘model’ species N. sylvestris, a rise, approximately 2-fold, in leaf nicotine levels was observed several days after a 4–5-fold increase in the transcript levels of all genes in the roots. In contrast, leaf tissues of the ornamental tobacco N. alata showed very low levels of any pyridine alkaloid, even when analysed 1 week after wounding, correlating with a general lack of transcript abundance representing any of these genes in leaves or roots following foliage damage. However, addition of methyl jasmonate to cultured roots of N. alata did produce elevated levels of nicotine and anatabine raising the possibility that components of the leaf–root wound signalling system in N. alata are different from those in N. sylvestris. Wounding of the tree tobacco N. glauca, was followed by a 2-fold increase in anabasine levels several days later. This increase followed a large rise in transcript levels of ODC, QPT and A622, though not PMT, in wounded leaves, but not in non-wounded leaves or roots. These data support the hypothesis that N. glauca is able to produce increased anabasine levels following wounding in its foliage, setting it apart from N. sylvestris where induced alkaloid production takes place in roots. We discuss the possibility that increased transcript levels detected by ODC and A622 probes play important roles in anabasine synthesis in N. glauca.
530. P. Singh et al., “Addressing Challenges to Enhance the Bioactives of Withania Somnifera through Organ, Tissue, and Cell Culture Based Approaches,” BioMed Research International, vol. 2017, p. e3278494, Feb. 2017. doi: 10.1155/2017/3278494. Abstract
     Withania somnifera is a highly valued medicinal plant in traditional home medicine and is known for a wide range of bioactivities. Its commercial cultivation is adversely affected by poor seed viability and germination. Infestation by various pests and pathogens, survival under unfavourable environmental conditions, narrow genetic base, and meager information regarding biosynthesis of secondary metabolites are some of the other existing challenges in the crop. Biotechnological interventions through organ, tissue, and cell culture provide promising options for addressing some of these issues. In vitro propagation facilitates conservation and sustainable utilization of the existing germplasms and broadening the genetic base. It would also provide means for efficient and rapid mass propagation of elite chemotypes and generating uniform plant material round the year for experimentation and industrial applications. The potential of in vitro cell/organ cultures for the production of therapeutically valuable compounds and their large-scale production in bioreactors has received significant attention in recent years. In vitro culture system further provides distinct advantage for studying various cellular and molecular processes leading to secondary metabolite accumulation and their regulation. Engineering plants through genetic transformation and development of hairy root culture system are powerful strategies for modulation of secondary metabolites. The present review highlights the developments and sketches current scenario in this field.
531. S. Singh, B. S. Tanwer, and M. Khan, “Callus Induction and in Vivo and in Vitro Comparative Study of Primary Metabolites of Withania Somnifera,” p. 6, 2011. Abstract
     In present study callus was raised from the leaf explant of Withania somnifera. Maximum callus was obtained on MS medium supplemented with 2, 4- D 1.0 mg/liter and IBA 0.5 mg/liter. The callus and different plant parts were used for primary metabolite quantification and antioxidant activity. Maximum soluble sugars found in callus, maximum amount of starch, protein and phenolic contents were found in stem and maximum lipid found in leaf.
532. V. Singh, B. Singh, R. Joshi, P. Jaju, and P. K. Pati, “Changes in the Leaf Proteome Profile of Withania Somnifera (L.) Dunal in Response to Alternaria Alternata Infection,” PLOS ONE, vol. 12, no. 6, p. e0178924, Jun. 2017. doi: 10.1371/journal.pone.0178924. Abstract
     Withania somnifera is a high value medicinal plant which is used against large number of ailments. The medicinal properties of the plant attributes to a wide array of important secondary metabolites. The plant is predominantly infected with leaf spot pathogen Alternaria alternata, which leads to substantial biodeterioration of pharmaceutically important metabolites. To develop an effective strategy to combat this disease, proteomics based approach could be useful. Hence, in the present study, three different protein extraction methods tris-buffer based, phenol based and trichloroacetic acid-acetone (TCA-acetone) based method were comparatively evaluated for two-dimensional electrophoresis (2-DE) analysis of W. somnifera. TCA-acetone method was found to be most effective and was further used to identify differentially expressed proteins in response to fungal infection. Thirty-eight differentially expressed proteins were identified by matrix assisted laser desorption/ionization time of flight-mass spectrometry (MALDI TOF/TOF MS/MS). The known proteins were categorized into eight different groups based on their function and maximum proteins belonged to energy and metabolism, cell structure, stress and defense and RNA/DNA categories. Differential expression of some key proteins were also crosschecked at transcriptomic level by using qRT-PCR and were found to be consistent with the 2-DE data. These outcomes enable us to evaluate modifications that take place at the proteomic level during a compatible host pathogen interaction. The comparative proteome analysis conducted in this paper revealed the involvement of many key proteins in the process of pathogenesis and further investigation of these identified proteins could assist in the discovery of new strategies for the development of pathogen resistance in the plant.
533. S. Singh, L. M. S. Palni, and D. S. Letham, “Cytokinin Biochemistry in Relation to Leaf Senescence V. Endogenous Cytokinin Levels and Metabolism of Zeatin Riboside in Leaf Discs from Green and Senescent Tobacco (Nicotiana Rustica) Leaves,” Journal of Plant Physiology, vol. 139, no. 3, pp. 279–283, Jan. 1992. doi: 10.1016/S0176-1617(11)80337-8. Abstract
     The cytokinin (bases and ribosides) content of leaf discs derived from green and senescent leaves of Nicotiana rustica was analyzed by radioimmunoassay. Zeatin was identified as the major base and dihydrozeatin riboside as the main riboside. Leaf discs from expanded green (presenescent) leaves contained three times higher zeatin levels than those from partly-senescent leaves. The uptake and metabolic fate of [3H]-zeatin riboside supplied to discs from tobacco leaves of different maturities was also investigated. The uptake of zeatin riboside declined with an increase in leaf maturity and senescence. Zeatin riboside was rapidly inactivated mainly by side-chain cleavage. Leaf discs excised from upper green leaves metabolized zeatin riboside into inactive compounds more rapidly than did the basal senescent leaves. It is suggested that higher cytokinin levels in discs from green leaves (in comparison to senescent leaves) may result from higher uptake of xylem cytokinins and/or in situ biosynthesis of cytokinins, rather than a more rapid degradation of zeatin-type cytokinins.
534. S. Singh, D. S. Letham, and L. M. S. Palni, “Effect of Growth Substances and Mineral Nutrients on Cytokinin Levels and Senescence of Tobacco(Nicotiana Rustica)Leaves,” in Progress in Plant Growth Regulation: Proceedings of the 14th International Conference on Plant Growth Substances, Amsterdam, 21–26 July, 1991, C. M. Karssen, L. C. van Loon, and D. Vreugdenhil, Eds. Dordrecht: Springer Netherlands, 1992, pp. 628–634. doi: 10.1007/978-94-011-2458-4_76. Abstract
     Several tobacco species exhibit a marked sequential leaf senescence in which basal leaves senesce first and then the senescence progresses towards apical leaves. The role of plant hormones, especially cytokinins, in the control of leaf senescence has been reviewed extensively (Van Staden et al. 1988). Cytokinins have been regarded as more effective senescence retardants than other growth substances because of their dramatic and pronounced senescence retarding activity when applied to diverse species (Noodén and Leopold, 1978).
535. S. Singh, \relax R. K. Vishwakarma, R. J. S. Kumar, P. D. Sonawane, Ruby, and B. M. Khan, “Functional Characterization of a Flavonoid Glycosyltransferase Gene from Withania Somnifera (Ashwagandha),” Applied Biochemistry and Biotechnology, vol. 170, no. 3, pp. 729–741, Jun. 2013. doi: 10.1007/s12010-013-0230-2. Abstract
     Glycosylation of flavonoids is mediated by family 1 uridine diphosphate (UDP)-dependent glycosyltransferases (UGTs). Until date, there are few reports on functionally characterized flavonoid glycosyltransferases from Withania somnifera. In this study, we cloned the glycosyltransferase gene from W. somnifera (UGT73A16) showing 85–92 % homology with UGTs from other plants. UGT73A16 was expressed as a His6-tagged fusion protein in Escherichia coli. Several compounds, including flavonoids, were screened as potential substrates for UGT73A16. HPLC analysis and hypsochromic shift indicated that UGT73A16 transfers a glucose molecule to several different flavonoids. Based on kinetic parameters, UGT73A16 shows more catalytic efficiency towards naringenin. Here, we explored UGT73A16 of W. somnifera as whole cell catalyst in E. coli. We used flavonoids (genistein, apigenin, kaempferol, naringenin, biochanin A, and daidzein) as substrates for this study. More than 95 % of the glucoside products were released into the medium, facilitating their isolation. Glycosylation of substrates occurred on the 7- and 3-hydroxyl group of the aglycone. UGT73A16 also displayed regiospecific glucosyl transfer activity towards 3-hydroxy flavone compound, which is the backbone of all flavonols and also for a chemically synthesized compound, not found naturally. The present study generates essential knowledge and molecular as well as biochemical tools that allow the verification of UGT73A16 in glycosylation.
536. V. Singh et al., “Leaf Spot Disease Adversely Affects Human Health-Promoting Constituents and Withanolide Biosynthesis in Withania Somnifera (L.) Dunal,” Journal of Applied Microbiology, vol. 122, no. 1, pp. 153–165, 2017. doi: 10.1111/jam.13314. Abstract
     Aims The present work investigates the implication of leaf spot disease on the antioxidant potential and commercial value of pharmaceutically important constituents of Withania somnifera, a high-valued medicinal plant. Methods and Results Leaf spot disease was induced in W. somnifera by inoculating Alternaria alternata (Fr.) Keiss. pathogen. Total polyphenolic content and antioxidant potential showed a significant decrease during leaf spot disease. Evaluation of pharmaceutically active constituents withaferin A, withanone and withanolide A utilizing high-performance liquid chromatography showed a significant decrease in diseased samples as compared to healthy ones. Quantitative expression of major genes involved in withanolide biosynthesis also showed down-regulation in diseased samples. Alterations in the ultra-structure of chloroplasts were also analysed under transmission electron microscopy to get a better insight into the changes of withanolide biosynthesis in leaf during disease infestation. Conclusions The present work suggests that when the pathogenic fungus invades the host plants, it evokes multiple responses, which could be studied at various levels. The knowledge gained from this work will provide appropriate rationale for controlling the bio-deterioration of the pharmaceutically active metabolites in W. somnifera and development of suitable strategies against leaf spot disease. Significance and Impact of the Study This is the first study to investigate the effect of leaf spot disease on the human health-promoting constituents and withanolide biosynthesis in this high-valued medicinal plant.
537. A. Singh, R. Gupta, M. Srivastava, M. M. Gupta, and R. Pandey, “Microbial Secondary Metabolites Ameliorate Growth, in Planta Contents and Lignification in Withania Somnifera (L.) Dunal,” Physiology and Molecular Biology of Plants, vol. 22, no. 2, pp. 253–260, Apr. 2016. doi: 10.1007/s12298-016-0359-x. Abstract
     In the present investigation, metabolites of Streptomyces sp. MTN14 and Trichoderma harzianum ThU significantly enhanced biomass yield (3.58 and 3.48 fold respectively) in comparison to the control plants. The secondary metabolites treatments also showed significant augmentation (0.75–2.25 fold) in withanolide A, a plant secondary metabolite. Lignin deposition, total phenolic and flavonoid content in W. somnifera were maximally induced in treatment having T. harzianum metabolites. Also, Trichoderma and Streptomyces metabolites were found much better in invoking in planta contents and antioxidants compared with their live culture treatments. Therefore, identification of new molecular effectors from metabolites of efficient microbes may be used as biopesticide and biofertilizer for commercial production of W. somnifera globally.
538. R. Singh et al., “Modulations in Primary and Secondary Metabolic Pathways and Adjustment in Physiological Behaviour of Withania Somnifera under Drought Stress,” Plant Science, vol. 272, pp. 42–54, Jul. 2018. doi: 10.1016/j.plantsci.2018.03.029. Abstract
     In general medicinal plants grown under water limiting conditions show much higher concentrations of secondary metabolites in comparison to control plants. In the present study, Withania somnifera plants were subjected to water stress and data related to drought tolerance phenomenon was collected and a putative mechanistic concept considering growth responses, physiological behaviour, and metabolite content and gene expression aspects is presented. Drought induced metabolic and physiological responses as well as drastic decrease in CO2 uptake due to stomatal limitations. As a result, the consumption of reduction equivalents (NADPH2+) for CO2 assimilation via the calvin cycle declines significantly resulting in the generation of a large oxidative stress and an oversupply of antioxidant enzymes. Drought also results in the shifting of metabolic processes towards biosynthetic activities that consume reduction equivalents. Thus, biosynthesis of reduced compounds (isoprenoids, phenols and alkaloids) is enhanced. The dynamics of various metabolites have been discussed in the light of gene expression analysis of control and drought treated leaves. Gene encoding enzymes of pathways leading to glucose, fructose and fructan production, conversion of triose phosphates to hexoses and hexose phosphorylation were up-regulated in the drought stressed leaves. The down-regulated Calvin cycle genes were co-ordinately regulated with the down-regulation of chloroplast triosephosphate/phosphate translocator, cytoplasmic fructose-1,6-bisphosphate aldolase and fructose bisphosphatase. Expression of gene encoding Squalene Synthase (SQS) was highly upregulated under drought stress which is responsible for the diversion of carbon flux towards withanolides biosynthesis from isoprenoid pathway.
539. A. K. Singh, R. Varshney, M. Sharma, S. S. Agarwal, and K. C. Bansal, “Regeneration of Plants from Alginate-Encapsulated Shoot Tips of Withania Somnifera (L.) Dunal, a Medicinally Important Plant Species,” Journal of Plant Physiology, vol. 163, no. 2, pp. 220–223, Feb. 2006. doi: 10.1016/j.jplph.2005.06.013. Abstract
     A protocol was developed for plant regeneration from encapsulated shoot tips collected from in vitro proliferated shoots of Withania somnifera. The best gel composition was achieved using 3% sodium alginate and 75mM CaCl2·2H2O. The maximum percentage response (87%) for conversion of encapsulated shoot tips into plantlets was achieved on MS medium supplemented with 0.5mg/l IBA after 5 weeks of culture. The conversion of encapsulated shoot tips into plantlets also occurred when calcium alginate beads having entrapped propagules were directly sown in autoclaved soilrite moistened with 14‐MS salts.
540. A. Singh, K. Lawrence, S. Pandit, and R. S. Lawrence, “Response of Leaves, Stems and Roots of Withania Somnifera to Copper Stress.,” International Journal of Plant, Animal and Environmental Sciences, vol. 4, no. 3, pp. 60–67, 2014. https://www.cabdirect.org/cabdirect/abstract/20143288363. Abstract
     The effects of Cu+2 stress (0, 10, 20, 50, 100 and 200 mM as CuSO4.5H2O) on growth and biochemical parameters of Withania somnifera L. Dunal, grown in a field pot experiment were studied. Increasing Cu+2 concentrations led to decreased stem length, root elongation and leaf area. Copper (Cu+2) stress decreased chlorophyll and carotenoids content in leaves and stems as compared to controls; this...
541. G. Singh, M. Tiwari, S. P. Singh, S. Singh, P. K. Trivedi, and P. Misra, “Silencing of Sterol Glycosyltransferases Modulates the Withanolide Biosynthesis and Leads to Compromised Basal Immunity of Withania Somnifera,” Scientific Reports, vol. 6, no. 1, p. 25562, May 2016. doi: 10.1038/srep25562. Abstract
     Sterol glycosyltransferases (SGTs) catalyse transfer of glycon moiety to sterols and their related compounds to produce diverse glyco-conjugates or steryl glycosides with different biological and pharmacological activities. Functional studies of SGTs from Withania somnifera indicated their role in abiotic stresses but details about role under biotic stress are still unknown. Here, we have elucidated the function of SGTs by silencing SGTL1, SGTL2 and SGTL4 in Withania somnifera. Down-regulation of SGTs by artificial miRNAs led to the enhanced accumulation of withanolide A, withaferin A, sitosterol, stigmasterol and decreased content of withanoside V in Virus Induced Gene Silencing (VIGS) lines. This was further correlated with increased expression of WsHMGR, WsDXR, WsFPPS, WsCYP710A1, WsSTE1 and WsDWF5 genes, involved in withanolide biosynthesis. These variations of withanolide concentrations in silenced lines resulted in pathogen susceptibility as compared to control plants. The infection of Alternaria alternata causes increased salicylic acid, callose deposition, superoxide dismutase and H2O2 in aMIR-VIGS lines. The expression of biotic stress related genes, namely, WsPR1, WsDFS, WsSPI and WsPR10 were also enhanced in aMIR-VIGS lines in time dependent manner. Taken together, our observations revealed that a positive feedback regulation of withanolide biosynthesis occurred by silencing of SGTLs which resulted in reduced biotic tolerance.
542. G. Singh et al., “Sterol Glycosyltransferases Required for Adaptation of Withania Somnifera at High Temperature,” Physiologia Plantarum, vol. 160, no. 3, pp. 297–311, 2017. doi: 10.1111/ppl.12563. Abstract
     Heat is a major environmental stress factor that confines growth, productivity, and metabolism of plants. Plants respond to such unfavorable conditions through changes in their physiological, biochemical and developmental processes. Withania somnifera, an important medicinal plant, grows in hot and dry conditions, however, molecular mechanisms related to such adaptive properties are not known. Here, we elucidated that members of the sterol glycosyltransferases (SGT) gene family play important roles in the survival of W. somnifera under adverse conditions through maintaining the integrity of the membrane. SGTs are enzymes involved in sterol modifications and participate in metabolic flexibility during stress. Silencing of WsSGT members, for instance WsSGTL1, WsSGTL2 and WsSGTL4, was inimical for important physiological parameters, such as electron transport rate, photochemical quantum yield, acceptor side limitation, non-photochemical quenching (NPQ), Fv/Fm and net photosynthetic rate, whereas stomatal conductance, transpiration rate and dark respiration rates (Rds) were increased. Decreased NPQ and increased Rds helped to generate significant amount of ROS in the Wsamisgt lines. After heat stress, H2O2, lipid peroxidation and nitric oxide production increased in the Wsamisgt lines due to high ROS generation. The expression of HSPs in Wsamisgt lines might be involved in regulation of physiological processes during stress. We have also observed increased proline accumulation which might be involved in restricting water loss in the Wsamisgt lines. Taken together, our observations revealed that SGTL enzyme activity is required to maintain the internal damages of the cell against high temperature by maintaining the sterol vs sterol glycosides ratio in the membranes of W. somnifera.
543. G. Singh, N. Dodiya, A. Joshi, and C. Khatik, “Variability, Character Associations and Path Analysis in Ashwagandha (Withania Somnifera (L). Dunal) with Respect to Root Yield and Biochemical Aspects,” Jan. 2017. Abstract
     ABSTRACTBackground: The experimental study was carried out to know the genetic variability, characters association,interrelationship and cause and effect of various characters in ashwagandha with respect to dry root yield andbiochemical aspects.Methods: Twenty genotypes (including 3 checks) were evaluated for 13 traits (10 quantitative and 3 qualitative)and analysis of variance, correlation and path analysis were performed for the mean data.Results: All characters were found to be differing significantly among genotypes. Estimates of variability parametersrevealed that a high genotypic coefficient of variation (GCV) and phenotypic coefficient of variation (PCV) were foundfor total antioxidants content in root, dry root weight per plant, dry plant weight per plant and harvest index(%). High heritability (h2) was found for total crude fiber content in root, total antioxidants content in root, plantheight and number of secondary branches per plant. Total antioxidants content in root, dry plant weight and totalcrude fiber content in root were recorded with high genetic advance (GA). A high heritability coupled with highgenetic advance was found for total antioxidants and total crude fiber content in root. Root diameter, plant height,dry plant weight and days to 75% maturity were shown significant positive correlation with dry root yield perplant. Path coefficient analysis revealed that root diameter, days to75% maturity and plant height had shown highpositive and direct effect on dry root yield per plant.Conclusion: The heritable variability and estimates of variability can be used for crop improvement. Rootdiameter, days to 75% maturity, plant height and dry plant weight could be used to select high dry root yieldinggenotypes. Biochemical data (alkaloid and antioxidants content) will be useful to select genotypes of high medicinalvalue.Keywords: Variability, Correlation, Path analysis, Heritability (h2), Genetic advance (GA), Biochemical aspects Variability, Character Associations and Path Analysis in Ashwagandha (Withania somnifera (L). Dunal) with Respect to Root Yield and Biochemical Aspects.
544. M. Singh, P. Shah, H. Punetha, and S. Agrawal, “Varietal Comparison of Withanolide Contents in Different Tissues of Withania Somnifera (L.) Dunal (Ashwagandha),” International Journal of Life-Sciences Scientific Research, vol. 4, May 2018. doi: 10.21276/ijlssr.2018.4.3.3. Abstract
     Five varieties of Ashwagandha (Withania somnifera L. (Dunal) i.e. Chetak, Pratap, Nimithli, Poshita and Jawahar-20 were analysed for withanolide A and withaferin A. These components were evaluated from leaf, stem, root and seeds using HPLC. These components were present in all these tissues. However, their content varies from tissues to tissue and variety to variety viz (Chetak, Pratap, Nimithli, Poshita and Jawahar-20). Withaferin A and withanolide A contents were found to be highest in Poshita followed by Jawahar-20. Withaferin A and withanolide A contents were also evaluated and compared in field grown and in-vitro grown plants of Poshita and Jawahar-20. Callus derived from Poshita root showed highest withaferin A content however, withanolide A was found to be highest in callus derived from Jawahar-20 leaf. Thus this study indicates that promoting an elite variety through tissue culture with consistency in the withanolide contents can be a promising approach to meet the growing demand of Ashwagandha.
545. P. Singh, R. Guleri, and P. K. Pati, “In Vitro Propagation of Withania Somnifera (L.) Dunal,” in Protocols for In Vitro Cultures and Secondary Metabolite Analysis of Aromatic and Medicinal Plants, Second Edition, S. M. Jain, Ed. New York, NY: Springer, 2016, pp. 201–213. doi: 10.1007/978-1-4939-3332-7_14. Abstract
     Withania somnifera (L.) Dunal known as Ashwagandha is commonly used in traditional Indian medicine system. It possesses immense therapeutic value against a large number of ailments such as mental diseases, asthma, inflammation, arthritis, rheumatism, tuberculosis, and a variety of other diseases including cancer. The therapeutic potential of W. somnifera is due to the presence of secondary metabolites mainly, tropane alkaloids and withanolides (steroidal lactones). The growing realization of commercial value of the plant has initiated a new demand for in vitro propagation of elite chemotypes of Withania. Micropropagation which is an important tool for rapid multiplication requires optimization of number of factors such as nutrient medium, status of medium (solid and liquid), type of explant, and plant growth regulators. Similarly, an efficient and reproducible in vitro regeneration system which is a prerequisite for the development of genetic transformation protocol requires precise manipulation of various intrinsic and extrinsic factors.
546. A. K. Singh et al., “A WRKY Transcription Factor from Withania Somnifera Regulates Triterpenoid Withanolide Accumulation and Biotic Stress Tolerance through Modulation of Phytosterol and Defense Pathways,” New Phytologist, vol. 215, no. 3, pp. 1115–1131, 2017. doi: 10.1111/nph.14663. Abstract
     Withania somnifera produces pharmacologically important triterpenoid withanolides that are derived via phytosterol pathway; however, their biosynthesis and regulation remain to be elucidated. A jasmonate- and salicin-inducible WRKY transcription factor from W. somnifera (WsWRKY1) exhibiting correlation with withaferin A accumulation was functionally characterized employing virus-induced gene silencing and overexpression studies combined with transcript and metabolite analyses, and chromatin immunoprecipitation assay. WsWRKY1 silencing resulted in stunted plant growth, reduced transcripts of phytosterol pathway genes with corresponding reduction in phytosterols and withanolides in W. somnifera. Its overexpression elevated the biosynthesis of triterpenoids in W. somnifera (phytosterols and withanolides), as well as tobacco and tomato (phytosterols). Moreover, WsWRKY1 binds to W-box sequences in promoters of W. somnifera genes encoding squalene synthase and squalene epoxidase, indicating its direct regulation of triterpenoid pathway. Furthermore, while WsWRKY1 silencing in W. somnifera compromised the tolerance to bacterial growth, fungal infection, and insect feeding, its overexpression in tobacco led to improved biotic stress tolerance. Together these findings demonstrate that WsWRKY1 has a positive regulatory role on phytosterol and withanolides biosynthesis, and defense against biotic stress, highlighting its importance as a metabolic engineering tool for simultaneous improvement of triterpenoid biosynthesis and plant defense.
547. V. Singhal and P. Kumar, “Cytomixis during Microsporogenesis in the Diploid and Tetraploid Cytotypes of Withania Somnifera (L.) Dunal, 1852 (Solanaceae),” Comparative cytogenetics, vol. 2, pp. 85–92., Jan. 2008. Abstract
     Withania somnifera (L.) Dunal, 1852, popularly known as “Ashwagandha” shows considerable morphogenetic diversity in Indian populations. Both the diploid (n=12) and tetraploid (n=24) cytotypes presently reported in the populations from Punjab, Rajasthan and Himachal Pradesh show the phenomenon of cytomixis which is a first record for the species. The inter PMC (Pollen mother cell) transfer of chromatin material in the diploid cytotype is present in 40.76% PMCs involving 2-6 PMCs. However, among tetraploids frequency of cytomixis is much less (15.80-24.32%) and involves only 2-3 PMCs. Chromatin transfer is noticed during the first and second meiotic division in the diploid as compared with the tetraploid where it is observed only during the first meiotic division. The percentage of meiotic abnormalities such as chromosome stickiness, lagging of chromatin material during anaphases and telophases is also higher in the diploid compared with the tetraploid. The microsporogenesis is also abnormal in the diploid resulting into the formation of polyads and tetrads with micronuclei whereas it is normal in the tetraploid. Cytomixis also seems to have had a greater effect on the apparent pollen fertility in the diploid compared with the tetraploid. Furthermore, cytomixis have also resulted into pollen grains of three different sizes in the diploid and only of two sizes in the tetraploid.
548. A. Sinha, R. Jain, S. Kachhwaha, and S. L. Kothari, “Optimization of the Level of Micronutrient Copper in the Culture Medium Improves Shoot Bud Regeneration in Indian Ginseng [Withania Somnifera (L.) Dunal].,” National Academy Science Letters, vol. 33, no. 1/2, pp. 11–16, 2010. https://www.cabdirect.org/cabdirect/abstract/20113218518. Abstract
     The effect of copper sulphate (CuSO4) on the formation of shoot buds and roots from nodal segments of Withania somnifera was investigated. Nodal segments formed multiple shoots through bud break on MS medium with BAP (22 µM) and IAA (5.7 µM). Formation of shoot buds was positively influenced by the presence of higher levels of copper ions in the medium. Upto 1.9 fold increase in the number of...
549. A. S. Siriwardan, R. M. Dharmadasa, and K. Samarasing, “Distribution Of Withaferin A, an Anticancer Potential Agent, In Different Parts of Two Varieties of Withania Somnifera (L.) Dunal. Grown in Sri Lanka,” Pakistan Journal of Biological Sciences, vol. 16, no. 3, pp. 141–144, Jan. 2013. doi: 10.3923/pjbs.2013.141.144.
550. V. A. Sisson and R. F. Severson, “Alkaloid Composition of the Nicotiana Species,” Beitr. Tabakforsch. Int., vol. 14, pp. 327–339, Jan. 2015. doi: 10.2478/cttr-2013-0610. Abstract
     Plants from 64 Nicotiana species were sampled in the greenhouse in 1983 and 1984 and from the field in 1985 and 1986 for the purpose of developing a chemical profile of each species. Mature green leaves were harvested at flowering, freeze-dried and ground to pass a 1 mm screen prior to chemical analysis. In this study we report the type and amounts of nicotinoid alkaloids. Alkaloid values were determined by glass-capillary gas chromatography and were averaged over the two years for each environment. Both total alkaloids (mg g-1 dry weight) and the distribution (percentage composition) of nicotine, nornicotine, anabasine, and anatabine were highly correlated between years for greenhouse and field samples. Greenhouse and field data were highly correlated, although total-alkaloid levels were significantly higher from field-grown plants. All of the Nicotiana species tested contained a measurable alkaloid fraction (at least 10 µg g-1). There was a wide range in total-alkaloid levels with nearly a 200-fold difference among greenhouse-grown samples and a 400-fold difference among field-grown species. In general, total-alkaloid levels among the species were found to be relatively low. Eleven species were found to have a different alkaloid composition when green leaf and air-dried samples were compared. Evidence is presented for the association between alkaloid characteristics and the phylogenetic classification of the species in the genus.
551. G. Sivanandhan et al., “The Effect of Polyamines on the Efficiency of Multiplication and Rooting of Withania Somnifera (L.) Dunal and Content of Some Withanolides in Obtained Plants,” Acta Physiologiae Plantarum, vol. 33, no. 6, pp. 2279–2288, Nov. 2011. doi: 10.1007/s11738-011-0768-y. Abstract
     An efficient mass multiplication protocol was developed for Withania somnifera (L.) Dunal from nodal explants of field-grown plants on Murashige and Skoog medium (MS) supplemented with 6-benzyladenine (BA) [1.5 mg L−l], indole-3-acetic acid (IAA) [0.3 mg L−l] and with the addition of polyamine, spermidine (20 mg L−l) (shoot multiplication medium). A total of 46.4 shoots were obtained from nodal explants and they were elongated in the same medium in a culture duration of 6 weeks. The elongated shoots produced roots in MS medium fortified with putrescine (20 mg L−l) after 4 weeks, and all the rooted plants were successfully hardened and acclimatized with a survival rate of 100%. An average of 276 shoots (46 × 6) was produced when at least six nodal explants obtained from each of the 46 in vitro grown shoots were cultured by microcutting method in the same shoot multiplication medium. On an average, 12,696 plants could be produced from all the shoots (276 × 46) by microcuttings in a period of 7 months. HPLC revealed a significant increase in the quantities of withanolide A, withanolide B, withaferin A and withanone in the leaves, stems, and roots of in vitro regenerated plants compared to the field-grown parent plants. Ploidy analysis using flow cytometry revealed genetic stability of in vitro regenerated plants. This protocol will be useful for scale-up production of withanolides on commercial scale.
552. G. Sivanandhan et al., “Effect of Culture Conditions, Cytokinins, Methyl Jasmonate and Salicylic Acid on the Biomass Accumulation and Production of Withanolides in Multiple Shoot Culture of Withania Somnifera (L.) Dunal Using Liquid Culture,” Acta Physiologiae Plantarum, vol. 35, no. 3, pp. 715–728, Mar. 2013. doi: 10.1007/s11738-012-1112-x. Abstract
     The influence of cytokinins and culture conditions including medium volume, harvest time and elicitation with abiotic elicitors (SA/MeJ) have been studied for the optimal production of biomass and withanolides in the multiple shoot culture of Withania somnifera. Elicitation of shoot inoculum mass (2 g l−l FW) with SA at 100 μM in the presence of 0.6 mg l−l BA and 20 mg l−l spermidine for 4 h exposure time at the 4th week in 20 ml liquid medium recorded higher withanolides production (withanolides A [8.48 mg g−l DW], withanolides B [15.47 mg g−l DW], withaferin A [29.55 mg g−l DW] and withanone [23.44 mg g−l DW]), which were 1.14 to 1.18-fold higher than elicitation with MeJ at 100 μM after 5 weeks of culture. SA-elicited cultures did not exhibit much variation in biomass accumulation when compared to control. This cytokinin induces and SA-elicited multiple shoot culture protocol provides a potential alternative for the optimal production of biomass and withanolides utilizing liquid culture.
553. G. Sivanandhan, N. Selvaraj, A. Ganapathi, and M. Manickavasagam, “Effect of Nitrogen and Carbon Sources on in Vitro Shoot Multiplication, Root Induction and Withanolides Content in Withania Somnifera (L.) Dunal,” Acta Physiologiae Plantarum, vol. 37, no. 2, p. 12, Jan. 2015. doi: 10.1007/s11738-014-1758-7. Abstract
     Effect of nitrogen and carbon sources on multiple shoot regeneration and withanolides contents of Withania somnifera were evaluated. Inclusion of l-glutamine (20 mg/l) in medium fortified with the optimal levels of 1.5 mg/l 6-benzyladenine (BA) and 0.3 mg/l indole-3-acetic acid (IAA) resulted in the production of 58 shoots/explant. Sucrose at 4 % and 20 mg/l l-glutamine with idéal concentrations of BA and IAA improved shoot multiplication (62 shoots/explant) while 6 % sucrose enhanced withanolides contents in regenerated multiple shoots. All the shoots were rooted (26 roots/shoot) when cultured in MS medium amended with 15 mg/l ammonium nitrate, 2 mg/l IBA, and 2 % sucrose. The present study resulted in the production of 4.42-fold higher multiple shoots and 2.6-fold higher roots as well as enhancing the contents of all major withanolides (withaferin A, withanone, withanolide A and B) in regenerated plants when compared to previous reports on W. somnifera regeneration.
554. G. Sivanandhan, N. Selvaraj, A. Ganapathi, and M. Manickavasagam, “An Efficient Hairy Root Culture System for Withania Somnifera (L.) Dunal,” African Journal of Biotechnology, vol. 13, no. 43, 2014. doi: 10.4314/ajb.v13i43. Abstract
     Withania somnifera is an important aromatic medicinal plant and possesses wide array of pharmacological properties. In the present investigation, an improved version of hairy root culture system was developed by optimizing various transformation parameters such as type of explant, concentration of acetosyringone, Agrobacterium types and co-cultivation period. Between the leaf and cotyledon explants and two Agrobacterium rhizgenes strains (R1000 and A4) tested, leaf explants infected with R1000 and cocultured for five days on MS basal half strength medium in the presence of acetosyringone (100 µM) attained a higher frequency (88%) of hairy root induction. By adopting this protocol, we could utilize the hairy root culture for industrial scale production of withanolides.Keywords: Leaf explant, Agrobacterium rhizogenes, Withania somnifera, co-cultivation period, acetosyringone.African Journal of Biotechnology, Vol 13(43) 4141-4147
555. G. Sivanandhan, C. Arunachalam, N. Selvaraj, A. A. Sulaiman, Y. P. Lim, and A. Ganapathi, “Expression of Important Pathway Genes Involved in Withanolides Biosynthesis in Hairy Root Culture of Withania Somnifera upon Treatment with Gracilaria Edulis and Sargassum Wightii,” Plant Physiology and Biochemistry, vol. 91, pp. 61–64, Jun. 2015. doi: 10.1016/j.plaphy.2015.04.007. Abstract
     The investigation of seaweeds, Gracilaria edulis and Sargassum wightii extracts was carried out for the estimation of growth characteristics and major withanolides production in hairy root culture of Withania somnifera. The extract of G. edulis (50%) in MS liquid basal medium enabled maximum production of dry biomass (5.46 g DW) and withanolides contents (withanolide A 5.23 mg/g DW; withaferin A 2.24 mg/g DW and withanone 4.83 mg/g DW) in hairy roots after 40 days of culture with 48 h contact time. The obtained withanolides contents were significantly higher (2.32-fold–2.66-fold) in hairy root culture when compared to the control. RT PCR analysis of important pathway genes such as SE, SS, HMGR and FPPS exhibited substantial higher expression upon the seaweed extracts treatment in hairy root culture. This experiment would paw a platform for withanolides production in hairy root culture with the influence of sea weed extracts for pharmaceutical companies in the future.
556. G. Sivanandhan, N. Selvaraj, A. Ganapathi, and M. Manickavasagam, “Improved Production of Withanolides in Shoot Suspension Culture of Withania Somnifera (L.) Dunal by Seaweed Extracts,” Plant Cell, Tissue and Organ Culture (PCTOC), vol. 119, no. 1, pp. 221–225, Oct. 2014. doi: 10.1007/s11240-014-0521-5. Abstract
     The influence of Gracilaria edulis and Sargassum wightii extracts was investigated for the production of biomass and withanolides in the multiple shoot suspension culture of Withania somnifera. Supplementation of 40 % G. edulis extract in MS liquid medium for 24 h exposure time in the culture recorded the highest biomass accumulation [62.4 g fresh weight and 17.82 g dry weight (DW)] and withanolides production (withanolide A 0.76 mg/g DW; withanolide B 1.66 mg/g DW; withaferin A 2.80 mg/g DW and withanone 2.42 mg/g DW) after 5 weeks of culture, which were 1.45–1.58-fold higher than control culture. This naturally available G. edulis extract-treated multiple shoot suspension culture protocol offers a potential alternative for the optimum production of biomass and withanolides utilizing shake-flasks.
557. G. Sivanandhan et al., “Increased Production of Withanolide A, Withanone, and Withaferin A in Hairy Root Cultures of Withania Somnifera (L.) Dunal Elicited with Methyl Jasmonate and Salicylic Acid,” Plant Cell, Tissue and Organ Culture (PCTOC), vol. 114, no. 1, pp. 121–129, Jul. 2013. doi: 10.1007/s11240-013-0297-z. Abstract
     Withania somnifera, an important medicinal plant that possesses a variety of bioactive secondary metabolites collectively known as withanolides. Hairy roots with an initial inoculum mass of 5 g FW were elicited separately with methyl jasmonate (MeJ) and salicylic acid (SA) at various concentrations for different exposure times after 30 days of culture. Enhanced production of biomass (32.68 g FW and 5.54 g DW; 1.23-fold higher), withanolide A (132.44 mg/g DW; 58-fold higher), withanone (84.35 mg/g DW; 46-fold higher), and withaferin A (70.72 mg/g DW; 42-fold higher) were achieved from 40 day-old harvested hairy roots elicited with 150 μM SA for 4 h exposure time. The present study reports a higher production of withanolide A, withanone and withaferin A from the elicited-hairy roots of W. somnifera under optimal inoculum mass, harvest time, elicitor exposure time and its concentration. These results will be useful for biochemical and bioprocess engineering for the viable production of withanolides in hairy root culture.
558. G. Sivanandhan et al., “Optimization of Elicitation Conditions with Methyl Jasmonate and Salicylic Acid to Improve the Productivity of Withanolides in the Adventitious Root Culture of Withania Somnifera (L.) Dunal,” Applied Biochemistry and Biotechnology, vol. 168, no. 3, pp. 681–696, Oct. 2012. doi: 10.1007/s12010-012-9809-2. Abstract
     Adventitious root cultures derived from leaf derived callus of Withania somnifera (L.) Dunal were treated with methyl jasmonate and salicylic acid independently. Biomass accumulation, culture age, elicitation period, and culture duration were optimized for higher withanolides production in the two best-responding varieties collected from Kolli hills (Eastern Ghats) and Cumbum (Western Ghats) of Tamil Nadu, India. Between the two elicitors, salicylic acid (SA) improved the production of major withanolides (withanolide A, withanolide B, withaferin A, and withanone) as well as minor constituents (12-deoxy withastramonolide, withanoside V, and withanoside IV) in the Kolli hills variety. Treatment of root biomass (11.70 g FW) on 30-day-old adventitious root cultures with 150 μM SA for 4 h elicitor exposure period resulted in the production of 64.65 mg g−l dry weight (DW) withanolide A (48-fold), 33.74 mg g−l DW withanolide B (29-fold), 17.47 mg g−l DW withaferin A (20-fold), 42.88 mg g−l DW withanone (37-fold), 5.34 mg g−l DW 12-deoxy withastramonolide (nine fold), 7.23 mg g−l DW withanoside V (seven fold), and 9.45 mg g−l DW withanoside IV (nine fold) after 10 days of elicitation (40th day of culture) when compared to untreated cultures. This is the first report on the use of elicitation strategy on the significant improvement in withanolides production in the adventitious root cultures of W. somnifera.
559. G. Sivanandhan, G. K. Dev, J. Theboral, N. Selvaraj, A. Ganapathi, and M. Manickavasagam, “Sonication, Vacuum Infiltration and Thiol Compounds Enhance the Agrobacterium-Mediated Transformation Frequency of Withania Somnifera (L.) Dunal,” PLOS ONE, vol. 10, no. 4, p. e0124693, Apr. 2015. doi: 10.1371/journal.pone.0124693. Abstract
     In the present study, we have established a stable transformation protocol via Agrobacterium tumafacines for the pharmaceutically important Withania somnifera. Six day-old nodal explants were used for 3 day co-cultivation with Agrobacterium tumefaciens strain LBA4404 harbouring the vector pCAMIBA2301. Among the different injury treatments, sonication, vacuum infiltration and their combination treatments tested, a vacuum infiltration for 10 min followed by sonication for 10 sec with A. tumefaciens led to a higher transient GUS expression (84% explants expressing GUS at regenerating sites). In order to improve gene integration, thiol compounds were added to co-cultivation medium. A combined treatment of L-Cys at 100 mg/l, STS at 125 mg/l, DTT at 75 mg/l resulted in a higher GUS expression (90%) in the nodal explants. After 3 days of co-cultivation, the explants were subjected to three selection cycles with increasing concentrations of kanamycin [100 to 115 mg/l]. The integration and expression of gusA gene in T0 and T1 transgenic plants were confirmed by polymerase chain reaction (PCR), and Southern blott analysis. These transformed plants (T0 and T1) were fertile and morphologically normal. From the present investigation, we have achieved a higher transformation efficiency of (10%). Withanolides (withanolide A, withanolide B, withanone and withaferin A) contents of transformed plants (T0 and T1) were marginally higher than control plants.
560. G. Sivanandhan, N. Selvaraj, A. Ganapathi, and Y. P. Lim, “Up-Regulation of Squalene Synthase in Hairy Root Culture of Withania Somnifera (L.) Dunal Yields Higher Quantities of Withanolides,” Industrial Crops and Products, vol. 154, p. 112706, Oct. 2020. doi: 10.1016/j.indcrop.2020.112706. Abstract
     The roots of Indian ginseng, Withania somnifera (L.) Dunal (Solanaceae) are extensively used in ayurvedic and other traditional medicines. The plant produces a steroidal lactone, withanolide which has multiple health benefits. Squalene synthase plays a central role in regulating the withanolide biosynthetic pathway in W. somnifera. The present approach investigated the up-regulation of squalene synthase in withanolides biosynthesis pathway in hairy roots. The higher expression of squalene synthase was noticed in hairy root line Ws9 which exhibited withanolide A (2.82 mg/g DW), withanolide B (1.34 mg/g DW), withaferin A (1.83 mg/g DW) and withanone (1.97 mg/g DW) on the 40th day of culture with 1.08 to 1.25-times higher, respectively. Semi-quantitative and quantitative polymerase chain reaction confirmed the incorporation and expression of squalene synthase in cultured hairy roots.
561. I. Sivanesan and S. W. Park, “Optimizing Factors Affecting Adventitious Shoot Regeneration, in Vitro Flowering and Fruiting of Withania Somnifera (L.) Dunal,” Industrial Crops and Products, vol. 76, pp. 323–328, Dec. 2015. doi: 10.1016/j.indcrop.2015.05.014. Abstract
     The effects of plant growth regulators, sucrose and temperature on adventitious shoot regeneration, in vitro flowering and fruiting of Withania somnifera were investigated. The highest frequency of shoot regeneration (96%) with an average of 22.8 shoots per stem segment was obtained on Murashige and Skoog (MS) medium fortified with 2.0mgl−1 kinetin, 0.5mgl−1 α-naphthaleneacetic acid (NAA), 0.3mgl−1 gibberellic acid (GA3) and 40gl−1 sucrose. The greatest frequency of rooting (100%) with an average of 14.1 roots per shoot was obtained when the regenerated shoots were cultured on the half-strength MS medium containing 2.0mgl−1 indole-3-butyric acid (IBA) and 15gl−1 sucrose. Regenerated plantlets were successfully established in the soil with 98% survival rate. The acclimatized plants grew well, developed flowers that were morphologically similar to the donor plants. The highest frequency of flowering (88%) with an average of 8.3 flowers per shoot and the greatest frequency of fruiting (74.9%) with an average of 5.1 fruit per shoot were obtained when shoot tips were cultured on MS medium fortified with 0.3mgl−1 N6-benzyladenine (BA) and 60gl−1 sucrose, and maintained at 20°C. Seeds were separated from the in vitro formed mature fruits and placed on MS medium containing 0.3mgl−1 GA3 to test their viability. The percentage of seed germination was 66%. The developed in vitro culture protocol can be useful for large scale propagation, genetic transformation and plant breeding studies.
562. I. Sivanesan and K. Murugesan, “In Vitro Adventitious Shoot Formation from Leaf Explants of Withania Somnifera Dunal.,” PLANT CELL BIOTECHNOLOGY AND MOLECULAR BIOLOGY, pp. 163–166, Feb. 2005. https://ikppress.org/index.php/PCBMB/article/view/2050. Abstract
     An efficient protocol was developed for high frequency plant regeneration from leaf explants of Withania somnifera Dunal on Murashige and Skoog (1962) medium supplemented with different concentrations of Auxins and cytokinins. Frequency of shoot bud regeneration varied with dose of plant growth regulators in the medium. Highest frequency of shoot buds was obtained at a concentration of 1.0 mg/l Kin. In vitro rooting of microshoots were obtained by growing them in half strength MS medium supplemented with 2.0 mg/l IBA. Rooted plantlets were successfully transferred to the field, after acclimatization in the net house.
563. H. H. Smith and C. R. Smith, “Alkaloids in Certain Species and Interspecific Hybrids of Nicotiana,” Journal of Agricultural Research, vol. 65, no. 7, pp. 347–359, Oct. 1942. https://www.google.com/books/edition/Journal_of_Agricultural_Research/JOjUAAAAMAAJ?hl=en&gbpv=1&pg=PA347&printsec=frontcover. Abstract
     Although it has been known for many years that the main alkaloid in the cultivated species Nicotiana tabacum L. and N. rustica L. is nicotine (C10H14N2), comparitively little analystical chemical work has been done on the wild species of Nicotiana.
564. H. H. Smith and D. V. Abashian, “Chromatographic Investigations on the Alkaloid Content of Nicotiana Species and Interspecific Combinations,” American Journal of Botany, vol. 50, no. 5, pp. 435–447, 1963. doi: 10.1002/j.1537-2197.1963.tb07212.x. Abstract
     Smith, H. H., and D. V. Abashian. (Brookhaven Natl. Lab., Upton, N. Y.) Chromatographic investigations on the alkaloid content of Nicotiana species and interspecific combinations. Amer. Jour. Bot. 50(5): 435–447. Illus. 1963.—Alkaloid content was analyzed by paper-partition chromatography in the following Nicotianas: (1) 52 species representing all taxonomic sections and centers of geographical distribution; (2) 35 two-species combinations including 1 species of hybrid origin, 5 F1 interspecific hybrids, 24 amphiploids and 5 sesquiploids; (3) 14 three-species combinations including 6 hybrids between an amphiploid and a third species, and 4 different 3-species combinations with doubled chromosome number; (4) 2 four-species combinations. Most of the species contained predominantly 3 identified alkaloids: nicotine, nornicotine and anabasine. In addition, at least 6 other alkaloids, that are separable but which were not identified chemically, are characteristic of the genus and are present in varied but characteristic patterns in each species. In hybrid combinations the content with regard to the unidentified alkaloids was observed to be the same as both parents, the sum of the parental patterns, a new alkaloid appearing, or, most frequently, one or more of the parental alkaloids missing in the hybrid. Among the identified alkaloids, anabasine was most frequently dominant to the other 2 in hybrids, and nornicotine production was most frequently either dominant or partly dominant to nicotine production. No simple basis for the inheritance of alkaloidal contents was clearly evident, nor wore there clearly defined associations between phylogenetic position and the alkaloids observed. Studies on the 6 or more alkaloids, in addition to nicotine, nornicotine and anabasine, which are known to be characteristic of the genus, offer extensive materials for research on alkaloid biosynthesis.
565. H. H. Smith, “Fixing Transgressive Vigor in Nicotiana Rustica*.”
566. H. H. Smith and C. W. Bacon, “Increase Size and Nicotine Production in Selections from Intraspecific Hybrids of Nicotiana Rustica,” Journal of Agricultural Research, vol. 63, no. 8, pp. 457–467, Oct. 1941. https://www.google.com/books/edition/Journal_of_Agricultural_Research/BA8YSUlLYOgC?hl=en&gbpv=1&pg=PA457. Abstract
     At present nicotine is obtained from leaf stems and low-grade or scrap-leaf material of Nicotiana tabacum L. as a byproduct of the tobacco industry. However, with the possibility of an increasing demand for nicotine, because of its wider use in inseticides and for other purposes, there may be need for varieties of tobacco that can be grown profitably for the nicotine alone.
567. S. J. Smolenski, F. A. Crane, and R. F. Voigt, “Effects of the Ratio of Calcium to Potassium in the Nutrient Medium on the Growth and Alkaloid Production of Atropa Belladonna,” Journal of Pharmaceutical Sciences, vol. 56, no. 5, pp. 599–602, May 1967. doi: 10.1002/jps.2600560511. Abstract
     The increase of calcium/potassium ratios in nutrient solutions reduces growth of belladonna plants. This is evident in the reduced elongation of all stems, particularly the sympodial flowering branches, and in the fresh and dry weights of all plant parts. There appears to be a concurrent increase in the proportion of leaf to total plant at the expense of stem and root. The increase in calcium/potassium ratio results in higher yields of total nitrogen and alkaloids.
568. Z. Song, T. Li, and C. Gong, “A Review on Starch Changes in Tobacco Leaves during Flue-Curing,” Frontiers of Agriculture in China, vol. 3, no. 4, pp. 435–439, Dec. 2009. doi: 10.1007/s11703-009-0076-0. Abstract
     This article discusses a range of studies on starch changes in tobacco leaves during flue-curing. Important effects resulting from using different flue-curing equipment and techniques on starch content are presented. It concludes that future researches should be focused on the structure and physico-chemical properties of tobacco starch during the curing process.
569. F. Sporer, M. Sauerwein, and M. Wink, “Diurnal and Developmental Variation of Alkaloid Accumulation in Atropa Belladonna,” Acta Horticulturae, no. 331, pp. 381–386, Sep. 1993. doi: 10.17660/ActaHortic.1993.331.53. Abstract
     The production and distribution of tropane alkaloids in organs of wild Atropa belladonna plants was studied by HPLC. Two different growth stages were examined in June and July and in addition berries and seeds in September 1991. A seasonal variation in total alkaloid content and alkaloid patterns of these plant parts was observed. Hyoscyamine was the main product throughout. A diurnal variation of tropane alkaloid content could be determined. Two peaks were significant: maximal alkaloid yields were detected at early night and at early morning. In maturing seeds, the alkaloid content was highest in the afternoon.
570. A. Srivastava, A. K. Gupta, K. Shanker, M. M. Gupta, R. Mishra, and R. K. Lal, “Genetic Variability, Associations, and Path Analysis of Chemical and Morphological Traits in Indian Ginseng [Withania Somnifera (L.) Dunal] for Selection of Higher Yielding Genotypes,” Journal of Ginseng Research, vol. 42, no. 2, pp. 158–164, Apr. 2018. doi: 10.1016/j.jgr.2017.01.014. Abstract
     Background The study was carried out to assess the genetic variability present in ashwagandha and to examine the nature of associations of various traits to the root yield of the plant. Methods Fifty-three diverse genetic stocks of ashwagandha (Withania somnifera) were evaluated for 14 quantitative characteristics. Analysis of variance, correlation, and path coefficient analysis were performed using the mean data of 2 years. Results Analysis of variance revealed that the genotypes differed significantly for all characteristics studied. High heritability in conjunction with high genetic advance was observed for fresh root weight, 12 deoxywithastramonolide in roots, and plant height, which indicated that selection could be effective for these traits. Dry root weight has a tight linkage with plant height and fresh root weight. Further, in path coefficient analysis, fresh root weight, total alkaloid (%) in leaves, and 12 deoxywithastramonolide (%) in roots had the highest positive direct effect on dry root weight. Conclusion Therefore, these characteristics can be exploited to improve dry root weight in ashwagandha genotypes and there is also scope for the selection of promising and specific chemotypes (based on the alkaloid content) from the present germplasm.
571. C. Srivastava, “Taxonomic and Ethnobiological Status of Withania Somnifera (L.) Dunal: The Indian Ginseng,” Medicinal Plants - International Journal of Phytomedicines and Related Industries, vol. 1, no. 2, p. 129, 2009. doi: 10.5958/j.0975-4261.1.2.019.
572. M. Staaf, S. Back, A. Wiernik, I. Wahlberg, R. C. Long, and J. H. Young, “Formation of Tobacco-Specific Nitrosamines (TSNA) During Air-Curing: Conditions and Control,” Contributions to Tobacco & Nicotine Research, vol. 21, no. 6, pp. 321–330, Jun. 2005. doi: 10.2478/cttr-2013-0798. Abstract
     AbstractThe present review deals with studies performed during several consecutive years on the effect of air-curing on tobacco-specific nitrosamine (TSNA) formation and quality of tobacco. Temperature, relative humidity, water content and water activity data were collected during curing of dark tobacco in traditional air-curing barns and bulk-curing barns of different sizes, and chemical analysis of the cured tobacco were performed.
573. R. A. Steinberg, “Comparison of Daylength and Temperature Responses in Nicotiana and Its Taxonomic Sections,” American Journal of Botany, vol. 46, no. 4, pp. 261–268, 1959. doi: 10.1002/j.1537-2197.1959.tb07011.x. Abstract
     Steinberg, Robert A. (U.S.D.A., Beltsville, Md.) Comparison of daylength and temperature responses in Nicotiana and its taxonomic sections. Amer. Jour. Bot. 46(4): 261–268. Illus. 1959.—Fifty-seven of the sixty species of Nicotiana were grown in the greenhouse under long- and short-day regimes. Supplemental tungsten light of about 30 ft.-c. (bench) was used to extend natural illumination to 16 hr. daily. Short-day controls received natural illumination for 9.5–12 hr. daily from about September to March. Two temperature levels were also employed—one with temperature held uniformly at about 25°C. and the other with a day temperature of about 20°C. and a night temperature of about 10°C. Daylength behavior of the species ranged from long-day to day-neutral to short-day. All species were brought into flower and all, except N. acaulis and N. ameghinoi, formed viable seed in at least 1 of the 4 environments. A modified classification of photoperiodic flowering responses based on rapidity and not ability to flower was adopted to permit quantitative comparison of species responses to both daylength and temperature. Very few species flowered equally rapidly (day-neutral) in both the 10- and 16-hr. day-lengths. Temperature level caused modifications in response from long-day to day-neutral and vice versa, and from short-day to day-neutral and vice versa. Data for N. glauca and some other species would indicate that a greater spread between temperature levels could possibly lead to opposite classifications at upper and lower temperatures. Excellent agreement was found between daylength responses of the species and the 14 taxonomic sections of Goodspeed for the genus Nicotiana. Only 2 of the sections (Paniculatae and Undulatae) were heterogeneous in that both included short- and long-day species in the same section. The native habitat of all short-day species was South America. Certain of the species gave a compensatory response to variations in light duration and low temperature similar to that given by sugarbeets and other biennials. This phenomenon may therefore be of general occurrence. Use of a quantitative expression for photoperiodic flowering responses is proposed to avoid ambiguity. It is the quotient of days from sowing to first blossom on short-days divided by that on long-days. The value 0.620°C. (9–12) would read short-day at 20°C. with 9–12 hr. daylengths. Close agreement was found in daylength flowering ratios in successive tests in the greenhouse. The ratios alter under cold treatment with species susceptible to low-temperature stimulation or inhibition of blossoming.
574. R. A. Steinberg, “Production and Germination of Nicotiana Seed Grown Under Various Greenhouse Conditions,” Tobacco Science, vol. 216, pp. 4–46, 1960.
575. A. Steppuhn, K. Gase, B. Krock, R. Halitschke, and I. T. Baldwin, “Nicotine’s Defensive Function in Nature,” PLoS Biology, vol. 2, no. 8, p. e217, Aug. 2004. doi: 10.1371/journal.pbio.0020217. Abstract
     Plants produce metabolites that directly decrease herbivore performance, and as a consequence, herbivores are selected for resistance to these metabolites. To determine whether these metabolites actually function as defenses requires measuring the performance of plants that are altered only in the production of a certain metabolite. To date, the defensive value of most plant resistance traits has not been demonstrated in nature. We transformed native tobacco(Nicotiana attenuata) with a consensus fragment of its two putrescine N-methyl transferase (pmt) genes in either antisense or inverted-repeat (IRpmt) orientations. Only the latter reduced (by greater than 95%) constitutive and inducible nicotine. With D4-nicotinic acid (NA), we demonstrate that silencing pmt inhibits nicotine production, while the excess NA dimerizes to form anatabine. Larvae of the nicotine-adapted herbivore Manduca sexta (tobacco hornworm) grew faster and, like the beetle Diabrotica undecimpunctata, preferred IRpmt plants in choice tests. When planted in their native habitat, IRpmt plants were attacked more frequently and, compared to wild-type plants, lost 3-fold more leaf area from a variety of native herbivores, of which the beet armyworm, Spodoptera exigua, and Trimerotropis spp. grasshoppers caused the most damage. These results provide strong evidence that nicotine functions as an efficient defense in nature and highlights the value of transgenic techniques for ecological research., Transgenic plants confirm the role that nicotine has in protecting tobacco plants from predators in nature and demonstrates the power of transgenic tools in studying ecological interactions in the field
576. D. Stoeva and E. Zayova, “Positive Transgressive Segregation in Intervarietal Crosses of Tobacco (Nicotiana Tabacum L.) and Their Micropropagation,” vol. 3, p. 9, 2017. Abstract
     Three advanced oriental tobacco (Nicotiana tabacum L.) lines, derived from cross between three commercial cultivars tobacco and selected by their positive transgressive traits, are described. Transgressive segregation by agronomically significant quantitative traits such as plant height, number of leaves, vegetation period (time to maturity) and yield/plant was observed. Content of nicotine, reducing sugars, and total nitrogen was determined in the leaves. Compared to the better parent, the yield per 1000 m2 exceeded over 18.92% (Line 44), 30.81% (Line 58/2) and 56.76% (Line 59), respectively. In field conditions, the all three lines presented combined disease resistances to Tobacco Mosaic Virus (TMV), black shank disease (Phitophtora parasitika var. nicotianae) and downy mildew (Peronospora tabacina). Based on these results, we inferred that the transgressive selection can be used for development of valuable lines of interest of tobacco breeding. An effective micropropagation protocol of the transgressive tobacco lines was developed. As initial explants were used in vitro 30 days germinated seedling. The highest propagation rate was recorded on MS medium containing 1.0 mg L-1 6-Benzylaminopurine (BAP) and 0.1 mg L-1 Indole-3-Acetic Acid (IAA) after three weeks of culture. The plants were successfully rooted in ½ MS medium containing 0.1 mg L-1 Indole-3-Butyric Acid (IBA). The developed efficient simple protocol in the study could be used for obtaining of plants for breeding programs of N. tabacum. The studied lines may be released as new varieties or used to produce improved tobacco hybrids.
577. G. V. Subbaraju et al., “Ashwagandhanolide, a Bioactive Dimeric Thiowithanolide Isolated from the Roots of Withania Somnifera,” Journal of Natural Products, vol. 69, no. 12, pp. 1790–1792, Dec. 2006. doi: 10.1021/np060147p. Abstract
     A new dimeric withanolide, ashwagandhanolide (1), was isolated from the roots of an Ayurvedic medicinal herb, Withania somnifera. A detailed spectroscopic evaluation revealed its identity as a dimer with an unusual thioether linkage. Compound 1 displayed growth inhibition against human gastric (AGS), breast (MCF-7), central nervous system (SF-268), colon (HCT-116), and lung (NCI H460) cancer cell lines, with IC50 values in the range 0.43−1.48 μg/mL. In addition, it inhibited lipid peroxidation and the activity of the enzyme cyclooxygenase-2 in vitro.
578. I. Subhas and S. Bangale, “Comparative Study of Seed Germination and Percentage of Fungal Infection of Ashwagandha (Withania Somnifera (L.) Dunal.),” Research Journal of Recent Sciences, vol. 1, pp. 2277–2502, Aug. 2012. Abstract
     Ashwagandha (Withania somnifera (L.) Dual.), is a medicinal plant. Presently, there are five different varieties are under in cultivation. These varieties shows different rate of seed germination and fungal infection .These variation in percent of seed germination and fungal infection are investigated in the present work by blotter method.
579. S. Sumathi and P. R. Padma, “Antioxidant Status of Different Parts of Withania Somnifera.,” Plant Archives, vol. 8, no. 1, pp. 69–72, 2008. https://www.cabdirect.org/cabdirect/abstract/20083207529. Abstract
     Herbal medicine has come of age in the past decade, moving from the health food stores to being universally available. With the advent of modern medicine, humans started isolating the active components and used them as such or made it more effective for therapy. The use of plant compounds for pharmaceutical purposes has gradually increased in India. Of all the medicinal plants used in India,...
580. U. Supe, F. Dhote, and M. Roymon, “In Vitro Plant Regeneration of Withania Somnifera,” Plant Tissue Culture and Biotechnology, vol. 16, no. 2, pp. 111–115, Jan. 1970. doi: 10.3329/ptcb.v16i2.1112. Abstract
     Key words: Seed explants, Withania somnifera, MicropropagationDOI = 10.3329/ptcb.v16i2.1112Plant Tissue Cult. & Biotech. 16(2): 111-115, 2006 (December)
581. K.-i. Suzuki, Y. Yamada, and T. Hashimoto, “Expression of Atropa Belladonna Putrescine N-Methyltransferase Gene in Root Pericycle,” Plant and Cell Physiology, vol. 40, no. 3, pp. 289–297, Jan. 1999. doi: 10.1093/oxfordjournals.pcp.a029540.
582. T. Swathi, M. Padma, M. Rajkumar, and A. Sivasankar, “Effect of Sowing Time, Seed Rate and Harvesting Duration on Germination of Ashwagandha,” p. 2. Abstract
     The experiment was carried out at Herbal garden, Rajendranagar, Hyderabad during the 2009-10. The experiment was laid out in Completely Randomized Block Design with factorial concept comprising a total of 18 treatments replicated thrice with two sowing dates (August 15th and August 30th), three seed rates (10 kg/ha, 12 kg/ha and 14 kg/ha) and three harvesting durations (150 DAS, 180 DAS and 210 DAS). Early germination (9.22 days) and maximum germination percentage (43.27%) was recorded with August 15th sown crop when compared to August 30th sown crop.
583. M. Tabata and N. Hiraoka, “Variation of Alkaloid Production in Nicotiana Rustica Callus Cultures,” Physiologia Plantarum, vol. 38, no. 1, pp. 19–23, 1976. doi: 10.1111/j.1399-3054.1976.tb04851.x. Abstract
     Callus cultures have been established from the seed, root and leaf of Nicotiana rustica L. var. brasilia in a synthetic medium containing 1 μM 2,4-D and μM kinetin. These callus tissues behaved similarly not only in growth and organogenesis but also in nicotine production. The nicotine contents of callus cultures, which were in the order of 0.25–0.58% of dry weight during a few passages subsequent to callus induction, rapidly decreased to trace amounts in succeeding subcultures in association with the decline of the root-regenerating activity. On the other hand, free cells prepared from a callus tissue in the third passage developed into individual clones showing wide differences in growth and nicotine production. One of these clones gave rise to a relatively stable strain which is capable of producing nicotine at a high rate (0.29% of dry weight) in the absence of organization. The significance of these findings is discussed in connection with some results which have been reported for other callus cultures of Nicotiana species.
584. S. Takshak and S. B. Agrawal, “Effect of Ultraviolet-B Radiation on Biomass Production, Lipid Peroxidation, Reactive Oxygen Species, and Antioxidants in Withania Somnifera,” Biologia Plantarum, vol. 58, no. 2, pp. 328–334, Jun. 2014. doi: 10.1007/s10535-014-0390-0. Abstract
     The present study was aimed at understanding the effects of long term supplemental UV-B (3.6 kJ m−2 d−1) on biomass production, accumulation of reactive oxygen species, lipid peroxidation, and enzymatic antioxidants in leaves and roots of Withania somnifera (an indigenous medicinal plant). Under the UV-B treatment, a reduction in biomass and an increased malondialdehyde content (a characteristic of lipid peroxidation) were observed in both the shoots and roots. Amongst ROS, H2O2 content increased under UV-B in the leaves, whereas it decreased in the roots, and superoxide radical production rate decreased in both the plant parts. The activities of all enzymatic antioxidants tested (ascorbate peroxidase, catalase, glutathione reductase, peroxidase, polyphenol oxidase, and superoxide dismutase) increased under the UV-B treatment, the increase being greater in the roots.
585. S. Takshak and S. B. Agrawal, “Secondary Metabolites and Phenylpropanoid Pathway Enzymes as Influenced under Supplemental Ultraviolet-B Radiation in Withania Somnifera Dunal, an Indigenous Medicinal Plant,” Journal of Photochemistry and Photobiology B: Biology, vol. 140, pp. 332–343, Nov. 2014. doi: 10.1016/j.jphotobiol.2014.08.011. Abstract
     The present study aims to investigate the effects of supplemental ultraviolet B (3.6kJm−2day−1 above ambient) radiation on secondary metabolites and phenylpropanoid pathway enzymes of Withania somnifera under field conditions at 40, 70, and 100days after transplantation. Secondary metabolites’ (alkaloids, anthocyanins, carotenoids, flavonoids, lignin, phytosterols, saponins, and tannins) concentrations were analysed at the end of the treatments. Activities of phenylalanine ammonia lyase (PAL), cinnamyl alcohol dehydrogenase (CAD), 4-coumarate-CoA ligase (4CL), chalcone–flavanone isomerase (CHI), and dihydroflavonol reductase (DFR) were also determined. In treated plants, secondary metabolite-concentrations generally increased (higher concentrations being recorded in roots compared to leaves). Anomalies were recorded for lycopene in roots and phytosterols in leaves (all sampling ages); β-carotene declined in leaves at third sampling age. s-UV-B-treated plants depicted decrease in withanolide A content with concomitant increase in withaferin A (two major alkaloids analysed by HPLC) compared to their respective controls. Phenylpropanoid pathway enzyme-activities increased in leaves and roots under s-UV-B treatment, the latter showing greater increase. The study concludes that s-UV-B is a potent factor in increasing the concentrations of secondary metabolites and their biosynthetic pathway enzymes in W. somnifera.
586. M. Tariq, A. Akbar, Lataf-ul-Haq, and A. Khan, “Comparing Application Methods for Boron Fertilizer on the Yield and Quality of Tobacco (Nicotiana Tabacum L.),” Communications in Soil Science and Plant Analysis, vol. 41, no. 13, pp. 1525–1537, Jun. 2010. doi: 10.1080/00103624.2010.485234. Abstract
     The present study is based on the hypothesis that different methods of supplying boron (B) may have different effects on the yield and quality performance of tobacco crop. A field experiment was conducted to study the effect of different methods of B fertilization on the yield, quality, and leaf composition of flue-cured Virginia tobacco (cv. K-399) in 2007 at the Pakistan Tobacco Research Station, Mansehra. Three different methods (i.e., foliar spray, soil application, and root dipping at the rates of 0.25 kg ha–1, 1 kg ha–1, and 0.5 mg B L–1, respectively) in the form of boric acid along with a control (where no B was applied) were used in a randomized complete block design and replicated four times. Results revealed that different methods of B application significantly affected tobacco yield, quality, and nutrient uptake compared to the control. Maximum leaf area of 707 cm2, green leaf yield of 18553 kg ha–1, cured leaf yield of 2314 kg ha–1, grade index of 79%, nicotine content of 2.54%, and sugar content of 18.35% were noted in the treatment plot where B was applied as a foliar spray; however, in a few cases, there was no significant difference found among different methods of B application. Chloride and potassium contents were not significantly affected by any method of B fertilization. The B fertilization increased the concentration of this element in tobacco leaf, and a maximum concentration of B 48.55 mg kg–1 was noted in the foliar spray treatment. Moreover, fertilizer-use efficiency for different methods of B application revealed that foliar spray is more efficient as compared to soil application and root-dipping methods. Nutrient ratios to B such as potassium (K) / B and chloride (Cl) / B considerably decreased in tobacco leaf with increases in the concentration of B in leaves. These ratios provided some indication of the interrelationship of B with these nutrients in tobacco plants. Results also revealed that K/B and Cl/B ratios closely correlated with the grade index of tobacco leaf, and good grades of tobacco were found to be at a K/B ratio of 682 and a Cl/B ratio of 148 under the experimental conditions. The overall results indicated that the B foliar spray at the rate of 0.25 kg ha–1 significantly increased the yield, quality, and the nutrient uptake by the tobacco crop under the prevailing conditions and was more effective than other methods of B application.
587. M. Tariq, Z. Ahmad, S. A. Shah, Z. Gul, and S. A. Khan, “Phytochemical Analysis and Antibacterial Activity of Nicotiana Tabacum and Nicotiana Rustica,” RADS Journal of Biological Research & Applied Sciences, vol. 12, no. 1, pp. 59–63, Jul. 2021. doi: 10.37962/jbas.v12i1.317. Abstract
     Background: All over the world, natural products containing different secondary metabolites have been used for antibacterial purposes, and as folk medicines with significant effects. Amongst many different plants, Tobacco plants are cultivated all over the world, but natively belong to America. These plants contain variety of secondary metabolites and possess significant antibacterial activity. Objectives: To conduct phytochemical analysis and measure the antibacterial potential of Tobacco plants Nicotiana tabacum and  Nicotiana rustica  using their extracts. Methodology: Multiple chemical tests such as Wagner’s test and Fehling’s test were used to determine the presence of different types of secondary metabolites. Both plant species were also screened for their antibacterial activity using agar well diffusion method. Results: Phytochemical analysis of the extracts from both plant species i.e. Nicotiana tabacum and Nicotiana rustica  indicated the presence of secondary metabolites including tannins, alkaloids, terpenoids, saponins, steroids, and flavonoids. Significant antibacterial activity of both plant extracts was observed against Staphylococcus aureus, but not against Escherichia coli. Conclusion: It can be concluded that both plant extracts showed the presence of secondary metabolites, with significant inhibitory effect observed against Staphylococcus aureus, and no effect against Escherichia coli.
588. K. S. Teja, G. T. Sivaram, \relax K. M. Yuvaraj, L. Kadiri, and M. P. Rao, “Effect of Different Dates of Sowing, Organic Manures on Protein, Fiber and Alkaloid Content in Leaves and Roots of Ashwagandha (Withania Somnifera),” The Pharma Innovation Journal, vol. 11, no. 9, p. 3, 2022. Abstract
     An experiment was conducted at Dept. of PSMA Block, College of Horticulture, Dr. YSR Horticultural University, Anantharajupeta, to know the protein, fiber, alkaloid content in both leaves and roots of Ashwagandha. The experiment was laid out in Split Plot Design with a total of 16 treatments replicated thrice with four sowing dates (First fortnight of September, second fortnight of September, first fortnight of October and Second fortnight of October), three Organic manures (FYM, Vermicompost, Neem Cake) and NPK (40:60:20 kg ha-1). Results reveled that maximum protein, fiber, alkaloid content was recorded with September 15th sown crop when compared to other dates sown crop, when as organic manures shown all significant result in compared with RDF.
589. N. P. Teli, N. M. Patil, H. M. Pathak, S. R. Bhalsing, and V. L. Maheshwari, “Withania Somnifera (Ashwagandha): Regeneration through Meristem Culture,” Journal of Plant Biochemistry and Biotechnology, vol. 8, no. 2, pp. 109–111, Jul. 1999. doi: 10.1007/BF03263069. Abstract
     High frequency of callus induction was achieved from shoot tip explants (80%) of Withania somnifera (Hindi name-Ashwagandha) as compared to leaf explants (70%), on MS medium supplemented with IAA (56μM) and Kinetin (56μM). When hormone-free MS medium was fortified with vitamins [thiamine. HCI (3μM), nicotinic acid (40.7μM) and pyridoxine. HCI (24.3μM)], it was found suitable for complete plant regeneration through meristem culture.
590. N. S. Thakur, K. Verma, and R. Rana, “Growth and Yield Performance of Ashwagandha (Withania Somnifera) under Agroforestry,” Indian Journal of Agricultural Sciences, vol. 84, pp. 937–941, Aug. 2014. Abstract
     Investigations on effect of tree-crop combinations and nitrogen levels on growth, yield and withanolides content and yield of Withania somnifera L. Dunal were carried out during 2005-2006 and 2006-2007, in mid hills of Himachal Pradesh (India). W. somnifera was grown in association with Prunus persica (fruit), Grewia optiva, Morus alba (fodder) and Setaria sphacelata (grass). The distinctive tree-crop combinations (agroforestry systems) formed were Peach + Grewia + Setaria + W. somnifera, Peach + Morus + Setaria + W. somnifera, Peach + Setaria + W. somnifera, Grewia + Setaria + W. somnifera, Morus + Setaria + W. somnifera and W. somnifera as sole crop. Three nitrogen doses, viz. 40, 80 and 120 kg/ha were applied to W. somnifera. The plant height and leaf area of ashwagandha plants was not affected by tree-crop combinations. The nitrogen dose of 120 kg/ha resulted in maximum height (44.55 cm) and leaf area (17.76 cm 2/leaf). Leaf area index was maximum (0.97) for plants grown in association with Peach+Grewia+Setaria. Belowground, aboveground and total biomass yield was not affected significantly by various tree-crop combinations. It ranged from 3.23 to 3.91, 2.30 to 2.63 and 5.53 to 6.46 q/ha, respectively. The N dose 120 kg/ha gave significantly higher below, aboveground and total biomass yield to the tune of 5.23, 3.37 and 8.60 q/ha, respectively. Withanolide content in roots varied from 0.73 to o.79 per cent but was not affected due to different treecrop combinations and nitrogen doses.
591. The Homestead Adventure, “How to Harvest and Dry Ashwagandha.” Nov-2021. https://www.youtube.com/watch?v=br20BDL36SY. Abstract
     Ashwagandha is an Ayurvedic herb that has amazing health benefits for your body and mind. It can reduce cortisol levels, increase energy, improve blood sugar levels, balance hormones, reduce inflammation, reduce stress and anxiety, help with thyroid function, improve sleep, support adrenal function, improve brain function, increase muscle mass, boost libido, and so much more!
592. C. Thilip et al., “Elicitation of Withaferin-A in Hairy Root Culture of Withania Somnifera (L.) Dunal Using Natural Polysaccharides,” Biologia, vol. 74, no. 8, pp. 961–968, Aug. 2019. doi: 10.2478/s11756-019-00236-9. Abstract
     Withaferin-A (WA) is a major bioactive compound in the roots of Withania somnifera. It is reported to possess inhibitory effects against breast cancer, liver cancer, melanoma cancer and prostate cancer cells. We developed an elicitation based on the hairy root culture (HRC) protocol for the enhancement of the WA production. Agrobacterium rhizogenes strain R1000 was used for the establishment of hairy root culture. PCR analysis confirmed the integration of the root loci C (rol C) gene in transformed roots. Elicitation of the hairy roots with natural polysaccharides of sodium alginate (SA), k-carrageenan (kC) and chitosan (CH) at various concentrations in the most cases resulted in a higher amount of WA compound. Among the various elicitors, 100 mg L−1 CH resulted 4.03-fold increase in WA production than control. This study is the first report in W. somnifera hairy roots, where the natural polysaccharides are used as elicitors for the large-scale production of WA.
593. S. A. Thorat et al., “Differential Gene Expression and Withanolides Biosynthesis During in Vitro and Ex Vitro Growth of Withania Somnifera (L.) Dunal,” Frontiers in Plant Science, vol. 13, p. 917770, Jun. 2022. doi: 10.3389/fpls.2022.917770. Abstract
     Ashwagandha (Withania somnifera L. Dunal) is a medicinally important plant with withanolides as its major bioactive compounds, abundant in the roots and leaves. We examined the influence of plant growth regulators (PGRs) on direct organogenesis, adventitious root development, withanolide biosynthetic pathway gene expression, withanolide contents, and metabolites during vegetative and reproductive growth phases under in vitro and ex vitro conditions. The highest shooting responses were observed with 6-benzylaminopurine (BAP) (2.0 mg L–1) + Kinetin (KIN) (1.5 mg L–1) supplementation. Furthermore, BAP (2.0 mg L–1) + KIN (1.5 mg L–1) + gibberellic acid (GA3) (0.5 mg L–1) exhibited better elongation responses with in vitro flowering. Half-strength MS medium with indole-3-butyric acid (IBA) (1.5 mg L–1) exhibited the highest rooting responses and IBA (1.0 mg L–1) with highest fruits, and overall biomass. Higher contents of withaferin A (WFA) [∼8.2 mg g–1 dry weight (DW)] were detected in the reproductive phase, whereas substantially lower WFA contents (∼1.10 mg g–1 DW) were detected in the vegetative phase. Cycloartenol synthase (CAS) (P = 0.0025), sterol methyltransferase (SMT) (P = 0.0059), and 1-deoxy-D-xylulose-5-phosphate reductase (DXR) (P = 0.0375) genes resulted in a significant fold change in expression during the reproductive phase. The liquid chromatography-mass spectrometry (LC-MS) analysis revealed metabolites that were common (177) and distinct in reproductive (218) and vegetative (167) phases. Adventitious roots cultured using varying concentrations of indole-3-acetic acid (IAA) (0.5 mg L–1) + IBA (1.0 mg L–1) + GA3 (0.2 mg L–1) exhibited the highest biomass, and IAA (0.5 mg L–1) + IBA (1.0 mg L–1) exhibited the highest withanolides content. Overall, our findings demonstrate the peculiarity of withanolide biosynthesis during distinct growth phases, which is relevant for the large-scale production of withanolides.
594. A. F. Tiburcio, M. T. Piñol, and M. Serrano, “Effect of UV-C on Growth, Soluble Protein and Alkaloids in Nicotiana Rustica Plants,” Environmental and Experimental Botany, vol. 25, no. 3, pp. 203–210, Aug. 1985. doi: 10.1016/0098-8472(85)90004-8. Abstract
     The inlfluence of UV-C (254 nm) irradiation on growth, soluble protein and alkaloid content in Nicotiana rustica plants was studied. Plants irradiated daily for 1 min appeared to be retarded in growth, with thicker and shorter stems, and delayed in flowering recovered. There was a general increase plants. Fourteen days post-irradiation, plants apparently recovered. There was a general increase of soluble protein content and enhancement of nicotine levels (during the last stages of the growth cycle) in the irradiated leaves on a dry weight basis. Increased soluble protein was due to post-treatment recovery period. A ‘stimulatory’ effect on nicotine as produced by the UV-treatment. In irradiated leaves changes in soluble protein profile corresponded to an increased alkaloid content. Enhancement of the 74 kD bands and 38 kD bands (detected by SDS-PAGE) was comparable to changes reported in response to another stress condition. Increased alkaloid levels could be a plant response to stress induced by the UV treatment.
595. M. M. Tirani, M. M. Haghjou, and A. Ismaili, “Effect of bulk and nano zinc oxide on seed germination and growth indices in tobacco (Nicotiana tabacum L.) seedlings.,” Iranian Journal of Seed Science and Research, vol. 6, no. 3, 2019. https://www.cabdirect.org/cabdirect/abstract/20219901564. Abstract
     The applications of bulk and nano zinc oxides have been increased recently, which can cause widespread plant’s environment contaminations. This study investigated the effects of commercial bulk ZnO (<1,000 nm) and nano ZnO (25 nm) on Nicotiana tabacum. Seeds were treated with ten concentrations of two forms (bulk and nano) ZnO (0.04, 0.2, 2.5, 5, 10, 50, 100, 500, 1000 and 2000 ppm) and...
596. G. Tiwari et al., “Plant Regeneration from Mature Cotyledon, Embryo and Hypocotyl Explants of Withania Somnifera (L.) Dunal,” J Agric Tech, vol. 7, Jan. 2011. Abstract
     S. (2011) Plant regeneration from mature cotyledon, embryo and hypocotyl explants of Withania somnifera (L.) Dunal. Journal of Agricultural Technology 7(4):1023-1035. The explants with higher regeneration potential, mature cotyledon, embryo and hypocotyls explants of two genotypes of Withania somnifera namely: JA-20 and MWS-100 were cultured on MS basal media fortified with different concentrations and combinations of various auxins and cytokinins. Considering higher in vitro response, culture medium MS2D.5B (MS + 2.0 mg.l -1 2,4-D + 0.5 mg.l -1 BA + 30.0 g.l –1 sucrose + 7.5 g.l –1 agar) initiated calli in higher frequencies from cultured mature cotyledon and embryo explants, while the hypocotyl explant, induction medium MS2N.5Kn (MS + 2.0 mg.l -1 NAA + 0.5 mg.l -1 Kn + 30.0 g.l –1 sucrose + 7.5 g.l –1 agar) was proved superior for callus initiation. For formation of morphogenic calli and plantlet regeneration nutrient medium MS2B.5N (MS + 2.0 mg.l -1 BA + 0.5 mg.l -1 NAA + 30.0 g.l –1 sucrose + 7.5 g.l –1 agar) were investigated remarkably superior. Higher in vitro rooting response was achieved on rooting medium (MS + 2.0 mg.l -1 IBA + 15.0 g.l –1 sucrose + 7.5 g.l –1 agar). Higher in vitro morphogenic response was exhibited by explant mature embryo followed by explants mature cotyledon and hypocotyl. In terms of in vitro genotypic response genotype JA-20 was found significantly superior to MWS-100 for the most of the attributes investigated. Regenerated plantlets were established successfully in the field after hardening.
597. M. A. Toghyani, A. A. Ehsanpour, M. Shariati, and R. Emamzadeh, “The Studuy of Auxin and Cytokinin Changes and Somaclonal Variation in Regenerated Plant and Callus of Tobacco (Nicotiana Rustica L.),” Journal of Plant Research (Iranian Journal of Biology), vol. 29, no. 2, pp. 385–394, Aug. 2016. https://plant.ijbio.ir/article_871.html. Abstract
     Auxin and cytokinin are two phytohormones which are effective for various vital processes such as plant growth, development and coordinating the responses to the environmental stimuli. It has been argued that these two phytohormones can affect the process of organogenesis by balancing each other’s level. The MS medium including 2 mg/lit of BAP hormone and the MS culture medium including 1 mg/lit of NAA hormone and 0.05 mg/lit of Kenitine hormone were respectively used as media for regeneration and callus formation of the tobacco explants. The amounts of Auxin and cytokinin were estimated in the first, second and third months after regeneration in the new shoots and the first generation callus respectively. The results of the above analyses revealed the proportion of Auxin to cytokinin in the time span of three months and this proportion confirmed the quick and preventing effects of auxin in early stages of growth on synthesis and amount of cytokinin. However, the preventing effects of cytokinin on auxin are slow and are probably through managing and changing other developmental processes. In addition, the amount of auxin and cytokinin in callus was lower compared with the amount of it in the control plant. This can be due to the fact that the cells of the callus are not distinctive and are different from the cells of the leafs which have compartment and different sources for the synthesis of hormone, in fact, the only available hormonl source for calluses is the one provided in the culture medium.
598. S. Toki and T. Kameya, “The Use of Streptomycin Resistance and Chlorophyll Deficiency for Selection of Somatic Hybrids between Nicotiana Tabacum and N. Rustica,” Heredity, vol. 60, no. 1, pp. 1–5, Feb. 1988. doi: 10.1038/hdy.1988.1. Abstract
     To prove the usefulness for somatic hybridisation of a Nicotiana tabacum L. double mutant (SA), which is both streptomycin resistant and chlorophyll deficient, protoplasts obtained from SA suspension cultures were fused via the dextran method with protoplasts isolated from N. rustica L. mesophyll cells. Prospective somatic hybrids were selected for streptomycin resistance and the ability to produce chlorophyll in regenerated plants. By using this selection system, green plants were recovered from 5 colonies. The hybrid nature of these plants was confirmed by morphological studies and isoelectric focusing of the Fraction 1 protein (RuBP carboxylase). The present results show evidence for the potential use of the double mutant in studies of somatic hybridisation.
599. P. Towey and J. L. Jinks, “The Number of Phenotypes among the Conditioned Lines of Nicotiana Rustica,” Heredity, vol. 37, no. 3, pp. 357–364, Dec. 1976. doi: 10.1038/hdy.1976.100. Abstract
     Analysis of the cumulative data on the 24 lines produced by conditioning treatments in variety 16 of Nicotiana rustica shows that for the mature plant characteristics exemplified by final plant height, the lines fall into three groups, large, intermediate and small. In this respect they are in broad qualitative agreement with the three genotrophs induced in flax, although it is not yet certain which of the three, if any, in N. rustica corresponds to the original plastic genotroph. This classification, however, while of practical utility breaks down as more characters, particularly juvenile and developmental, are considered.
600. N. Tripathi, N. Saini, V. Mehto, S. Kumar, and S. Tiwari, “Assessment of Genetic Diversity among Withania Somnifera Collected from Central India Using RAPD and ISSR Analysis,” Medicinal and Aromatic Plant Science and Biotechnology, vol. 6, no. 1, pp. 33–39, 2012. https://www.researchgate.net/publication/235285945_Assessment_of_Genetic_Diversity_among_Withania_somnifera_Collected_from_Central_India_using_RAPD_and_ISSR_Analysis. Abstract
     Withania somnifera is a multipurpose plant of immense therapeutic value and wide geographic distribution exhibiting extensive phenotypic and chemical variability. Characterization of plants using molecular markers is an ideal approach for improvement and conservation of plant genetic resources. Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) molecular fingerprinting markers were employed as genetic markers to assay the genetic relationship of 16 accessions of W. somnifera collected from different locations of India. Randomly selected 25 decamer primers amplified 204 RAPD marker loci out of which 152 bands (74.5%) were polymorphic. Twenty eight ISSR primers were used to generate fingerprints and a total of 186 alleles were amplified, out of which 151 were polymorphic (81.1%). ISSR markers were more informative than the RAPD markers. Several ISSR markers amplified genotype-specific alleles that can be used for authentication and detection of adulteration in plant material. Similarity matrices were generated from RAPD and ISSR marker data separately using Jaccard’s coefficient and dendrograms were constructed based on UPGMA clustering. Genotypes were clustered into two groups and the grouping was similar for both marker systems.
601. S. Tripathi, R. S. Sangwan, B. Mishra, J. S. Jadaun, and N. S. Sangwan, “Berry Transcriptome: Insights into a Novel Resource to Understand Development Dependent Secondary Metabolism in Withania Somnifera (Ashwagandha),” Physiologia Plantarum, vol. 168, no. 1, pp. 148–173, 2020. doi: 10.1111/ppl.12943. Abstract
     Withania somnifera (Ashwagandha) is considered as Rasayana in Indian systems of medicine. This study reports a novel transcriptome of W. somnifera berries, with high depth, quality and coverage. Assembled and annotated transcripts for nearly all genes related with the withanolide biosynthetic pathway were obtained. Tissue-wide gene expression analysis reflected almost similar definitions for the terpenoid pathway in leaf, root and berry tissues with relatively higher abundance of transcripts linked to steroid, phenylpropanoid metabolism as well as flavonoid metabolism in berries. The metabolome map generated from the data embodied transcripts from 143 metabolic pathways connected together and mediated collectively by about 1792 unique enzyme functions specific to berry, leaf and root tissues, respectively. Transcripts specific to cytochrome p450 (CYP450), methyltransferases and glycosyltransferases were distinctively located in a tissue specific manner and exhibited a complex network. Significant distribution of transcription factor genes such as MYB, early light inducible protein (ELI), minichromosome maintenance1, agamous, deficiens and serum response factor (MADS) and WRKY etc. was observed, as the major transcriptional regulators of secondary metabolism. Validation of the assembly was ascertained by cloning WsELI, which has a light dependent regulatory role in development. Quantitative expression of WsELI was observed to be considerably modulated upon exposure to abiotic stress and elicitors. Co-relation of over-expression of WsELI, may provide new aspects for the functional role of ELI proteins in plants linked to secondary metabolism. The study offers the first comprehensive and comparative evaluation of W. somnifera transcriptome data between the three tissues and across other members of Solanaceae (Nicotiana, Solanum and Capsicum) with respect to major pathways and their metabolome regulation.
602. S. Tripathi, Y. Srivastava, R. S. Sangwan, and N. S. Sangwan, “In Silico Mining and Functional Analysis of AP2/ERF Gene in Withania Somnifera,” Scientific Reports, vol. 10, no. 1, p. 4877, Mar. 2020. doi: 10.1038/s41598-020-60090-7. Abstract
     Withania somnifera owing to its strong and remarkable stress tolerance property is a reliable candidate for the determination of genes involved in mechanism of adaption/tolerance of various stress conditions. 187 AP2/ERF gene related transcripts (GRTs) were identified during comprehensive search in W. somnifera transcriptome repertoire. Major hits in homology search were observed from the model plant Arabidopsis and members of Solanaceae family. Cloning, expression analysis of the gene and genetic transient transformation with the gene (WsAP2) were performed to predict its functional role in planta. Enhanced expression of some of the pathway genes for terpenoid biosynthesis was observed in transformed tissues in comparison to the control tissues. It is speculated that WsAP2 gene crucially regulates the expression of GGPPS gene in addition to the regulation of other important genes of terpenoid pathway via induction of expression of other genes such as HMGR, CAS, DXS and DXR. To the best of our knowledge, this is the first report representing detailed study of AP2/ERF gene family in W. somnifera. It is also suggested from the study that gene might have role in eliciting responses to combat stress and attribute the strong stress tolerant property associated with the plant.
603. S. Tripathi, R. S. Sangwan, L. K. Narnoliya, Y. Srivastava, B. Mishra, and N. S. Sangwan, “Transcription Factor Repertoire in Ashwagandha (Withania Somnifera) through Analytics of Transcriptomic Resources: Insights into Regulation of Development and Withanolide Metabolism,” Scientific Reports, vol. 7, no. 1, p. 16649, Nov. 2017. doi: 10.1038/s41598-017-14657-6. Abstract
     Transcription factors (TFs) are important regulators of cellular and metabolic functions including secondary metabolism. Deep and intensive RNA-seq analysis of Withania somnifera using transcriptomic databases provided 3532 annotated transcripts of transcription factors in leaf and root tissues, belonging to 90 different families with major abundance for WD-repeat (174 and 165 transcripts) and WRKY (93 and 80 transcripts) in root and leaf tissues respectively, followed by that of MYB, BHLH and AP2-ERF. Their detailed comparative analysis with Arabidopsis thaliana, Capsicum annum, Nicotiana tabacum and Solanum lycopersicum counterparts together gave interesting patterns. However, no homologs for WsWDR representatives, LWD1 and WUSCHEL, were observed in other Solanaceae species. The data extracted from the sequence read archives (SRA) in public domain databases were subjected to re-annotation, re-mining, re-analysis and validation for dominant occurrence of WRKY and WD-repeat (WDR) gene families. Expression of recombinant LWD1 and WUSCHEL proteins in homologous system led to enhancements in withanolide content indicating their regulatory role in planta in the biosynthesis. Contrasting expression profiles of WsLWD1 and WsWUSCHEL provided tissue-specific insights for their participation in the regulation of developmental processes. The in-depth analysis provided first full-spectrum and comparative characteristics of TF-transcripts across plant species, in the perspective of integrated tissue-specific regulation of metabolic processes including specialized metabolism.
604. C. Tsaltaki, M. Katsouli, T. Kekes, S. Chanioti, and C. Tzia, “Comparison Study for the Recovery of Bioactive Compounds from Tribulus Terrestris, Panax Ginseng, Gingko Biloba, Lepidium Meyenii, Turnera Diffusa and Withania Somnifera by Using Microwave-Assisted, Ultrasound-Assisted and Conventional Extraction Methods,” Industrial Crops and Products, vol. 142, p. 111875, Dec. 2019. doi: 10.1016/j.indcrop.2019.111875. Abstract
     Heat reflux (CONV), Soxhlet (SOX), microwave-assisted (MAE) and ultrasound-assisted (UAE) extraction methods, using ethanol, water and their mixtures, were studied for the recovery of bioactive compounds from Tribulus terrestris, Panax ginseng, Gingko biloba, Lepidium meyenii, Turnera diffusa and Withania somnifera. The study revealed that SOX method achieved extracts with the highest yieldTPC in terms of the total phenolic content (TPC) (66.65\,± 1.50%); MAE and UAE resulted also in extracts with high TPC and shorter treatment times compared to SOX. More specifically, the TPC values for MAE and UAE were 91.783\,± 0.026 mg GA/g dw and 81.113\,± 0.103 mg GA/g dw, respectively, the antioxidant activity (DPPH) was 63.577\,± 0.057 mg Trolox/g dw and 64.923 mg Trolox/g dw, and the yieldTPC was 60.82% and 61.89%, respectively. All were higher compared to those obtained by the CONV method. The use of pure ethanol as the extraction solvent, provided maximum TPC and similar results for DPPH and yieldTPC (%). Moreover, maximum TPC (183.70 mg GA/g dw) and DPPH (377.21 mg Trolox/g dw) values were achieved for T. diffusa. The HPLC analysis of T. terrestris, P. ginseng and G. biloba extracts showed that G. biloba contained its characteristic bioactive compound in the highest concentration (13.512 mg gingkolide/g herb).
605. T. C. Tso and R. N. Jeffrey, “Biochemical Studies on Tobacco Alkaloids. I. The Fate of Labeled Tobacco Alkaloids Supplied to Nicotiana Plants,” Archives of Biochemistry and Biophysics, vol. 80, no. 1, pp. 46–56, Jan. 1959. doi: 10.1016/0003-9861(59)90339-X. Abstract
     The N15-labeled alkaloids: nicotine, nornicotine, and anabasine, were each supplied through the roots of Nicotiana rustica var. brasilia plants in water culture. N15 and C14H3 double-labeled nicotine was supplied similarly to Nicotiana glauca to study the fate of these alkaloids in Nicotiana plants. A majority of the labeled alkaloid supplied was metabolized and the label appeared in other compounds. The major alkaloid found was characteristic of the experimental plant and was equally labeled with N15 in both rings, like the alkaloid supplied. Some of the N15 alkaloids supplied probably were converted to new alkaloids by a route more direct than through the general metabolic pool. The N15 to C14 ratio of the anabasine in the N. glauca plant was half that of the N15 methyl-C14 labeled nicotine supplied. Thus the anabasine was not formed by mere rearrangement of the nicotine molecule.
606. T. C. Tso, Physiology and Biochemistry of Tobacco Plants, First Edition. Stroudsburg, Pa: Dowden, Hutchinson & Ross, 1972.
607. R. Udayakumar et al., “Analysis of Genetic Variation among Populations of Withania Somnifera (L.) in South India Based on RAPD Markers.,” European Journal of Medicinal Plants, vol. 3, no. 2, pp. 266–280, 2013. doi: 10.9734/EJMP/2013/2801. Abstract
     Aim: The present study was carried out to analyze the genetic variations among 20 different populations of Withania somnifera (L.) Dunal collected from different habitats (locations) by RAPD analysis. Methodology: DNA was isolated from the fresh leaf samples collected from the field by Bernatsky and Tankley method. Isolated genomic DNA was purified by phenol: chloroform: isoamyl alcohol (25:24:1)...
608. R. Udayakumar, A. Ganapathi, K. SeiChang, and C. ChangWon, “Indirect Regeneration of Withania Somnifera from Nodal Explants.,” British Biotechnology Journal, vol. 4, no. 4, pp. 366–378, 2014. doi: 10.9734/BBJ/2014/7511. Abstract
     Withania somnifera is an important medicinal plant and used to cure many diseases. Indirect regeneration protocol for multiple shoots development was established using nodal explants of W. somnifera from 50-60 days old seedlings. The callus induction was observed from nodal explants, grown on Murashige and Skoog (MS) medium supplemented with various concentrations and combinations of...
609. P. Vaidyanathan and K. Francis, “Impact of Sildenafil Citrate on Tissue Culture of Tobacco (Nicotiana Tabaccum).,” Plant Cell Biotechnology and Molecular Biology, vol. 10, no. 1/2, pp. 49–56, 2009. https://www.cabdirect.org/cabdirect/abstract/20103330361. Abstract
     Plant growth is controlled by several biochemicals that are collectively known as plant growth substances or hormones or regulators. One among them is nitrogen monoxide commonly known as Nitric oxide (NO-). The enzymatic activity responsible for NO- generation is NOS, which uses L-arginine as substrate, promoting its oxidation, with NADPH and oxygen consumption, and yielding L-citrulline and NO....
610. B. Vajantha, M. Uma Devi, M. Chandini Patnaik, and M. Rajkumar, “Growth and Yield of Ashwagandha (Withania Somnifera L.) as Effected by INM and Panchagavya,” International Journal of Agricultural Sciences, vol. 8, no. 1, pp. 128–134, Jan. 2012. http://researchjournal.co.in/upload/assignments/8_128-134.pdf. Abstract
     Investigations were carried out for two years to study the effect of different levels of NPK (0, 50, 100 and 150%), organic manures (castor cake @ 2.5 t ha-1 and vermi-compost @ 1 t ha-1), panchagavya and bio fertilizers (Azospirillum and phosphorus solubilising bacteria each of 5 kg ha-1) on growth and root yield of medicinal crop Ashwagandha at College Farm, College of Agriculture, Hyderabad during Rabi 2007-08 and Kharif 2008 by using split plot design. During both the years at flowering and harvest, the highest dry matter production (3314 and 3083 kg ha-1 and 6204 and 5101 kg ha-1) and dry root yield (127 and 125 kg ha-1 and 348 and 333 kg ha-1, respectively) was recorded with conjunctive use of 150 per cent NPK with castor cake @ 2.5 t ha-1.
611. P. Van Dijk, F. A. Van Der Meer, and P. G. M. Piron, “Accessions of Australian Nicotiana Species Suitable as Indicator Hosts in the Diagnosis of Plant Virus Diseases,” Netherlands Journal of Plant Pathology, vol. 93, no. 2, pp. 73–85, Mar. 1987. doi: 10.1007/BF01998093. Abstract
     When screening the genusNicotiana for sensitive and differential hosts for a group of mechanically transmissible plant viruses with narrow host ranges, development of systemic symptoms was alsmost exclusively observed in species of three closely related sections of the subgenusPetunioides. These species wereN. miersii (sectionAcuminatae),N. bigelovii andN. clevelandii (Bigelovianae) andN. benthamiana, N. cavicola, N. ingulba, N. occidentalis, N. rosulata andN. rotundifolia (Suaveolentes). Except forN. benthamiana andN. clevelandii, which are already known for their large virus ranges, they are new experimental hosts that appeared very useful for detection of viruses and for differentiation of viruses that closely resemble each other in host range. Accessions of the same species often varied largely in local and systemic viral response. EspeciallyN. benthamiana-9,N. miersii-33 andN. occidentalis-37B (code numbers given by Tobacco Research Laboratory, Oxford, N.C., USA) are recommended for routine inoculation tests. The sensitiveSuaveolentes species mentioned are native to the arid parts of Australia. Collections of these species deserve attention in studies on virus diseases of unknown etiology where experimental hosts are lacking.
612. O. I. Varchenko, M. V. Kuchuk, M. F. Parii, and \relax Y. V. Symonenko, “Comparison of Gfp Gene Expression Levels after Agrobacterium-Mediated Transient Transformation of Nicotiana Rustica L. by Constructs with Different Promoter Sequences,” Cytology and Genetics, vol. 54, no. 6, pp. 531–538, Nov. 2020. doi: 10.3103/S0095452720060110. Abstract
     Promoters are key elements regulating gene expression levels, therefore their selection is an important step in genetic engineering research. The reporter gene gfp, which encodes green fluorescent protein (GFP), was transiently expressed in leaf tissues of Aztec tobacco Nicotiana rustica L. Compared to other species of the Nicotiana genus, Aztec tobacco has a large potential for expression of heterologous proteins, a large vegetative biomass, can be easily infiltrated, and is unpretentious in cultivation. Six genetic constructs were used with different promoter sequences: the 35S promoter of Cauliflower Mosaic Virus (35S CaMV), the double-enhanced 35S promoter (D35S CaMV), promoters of the RbcS1B and RbcS2B genes encoding the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) isolated from Arabidopsis thaliana (L.) Heynh., and promoters of the LHB1B1 and LHB1B2 genes from A. thaliana encoding chlorophyll a/b binding proteins. The gfp gene expression was detected visually, spectrofluorimetrically, and by protein content (Bradford assay) on the seventh day after infiltration. The highest level of expression was observed using the double-enhanced 35S promoter (D35S CaMV) and the lowest using the LHB1B1 gene promoter.
613. S. Varghese, R. Keshavachandran, B. Baby, and P. A. Nazeem, “Genetic Transformation in Ashwagandha ( Withania Somnifera (L.) Dunal) for Hairy Root Induction and Enhancement of Secondary Metabolites,” Journal of Tropical Agriculture, vol. 52, no. 1, pp. 47–53, Oct. 2014. http://jtropag.kau.in/index.php/ojs2/article/view/302. Abstract
     Genetic transformation was carried out in ashwagandha (Withania somnifera (L.) Dunal) using three different Agrobacterium rhizogenes strains viz., A4, ATCC 15834 and MTCC 2364, for inducing hairy roots. The explants such as hypocotyls, cotyledonary segments, leaf segments, shoot tips and nodal segments were used for genetic transformation. A4 and ATCC 15834 strains produced successful transformation and hairy (transformed) roots were induced from leaf segments and shoot tips. A4 strain produced transformation by direct inoculation of bacteria from single cell colonies as well as in the suspension form, but ATCC 15834 produced transformation only in the suspension form. Among the liquid media tested, half MS was found to be superior in promoting hairy root growth. The transformation was confirmed by PCR and dot blot analysis. A Thin Layer Chromatographic method was employed for withanolide estimation. The spot corresponding to withaferin A was observed under UV at 254 nm. Field root possessed more withaferin A followed by hairy roots and in vitro roots contained the least. Enhancement of secondary metabolite production was attempted through addition of osmoregulator, precursor feeding and elicitation. Withaferin A content in the hairy root biomass and the culture medium were estimated. The biotic elicitor Aspergillus homogenate (250 and 500 μl /125 ml) had a positive influence in the enhancement of secondary metabolites.
614. V. V. Velde and D. Lavie, “A Δ16-Withanolide in Withania Somnifera as a Possible Precursor for α-Side-Chains,” Phytochemistry, vol. 21, no. 3, pp. 731–733, Jan. 1982. doi: 10.1016/0031-9422(82)83176-2. Abstract
     The structure of a new naturally occurring steroidal lactone of the withanolide group isolated from Withania somnifera chemotype III has been elucidated as (20R,22R)-14α,2OαF-dihydroxy-1-oxowitha-2,5,16,24-tetraenolid. This compound is considered to be an intermediate in the biosynthesis of withanolide E, and is at the origin of the unusual α-oriented side-chain in this compound. The comparative composition of withanolides in different sub-chemotypes of III is provided.
615. G. Vembu and R. Singaravel, “Response of Ashwagandha (Withania Somnifera Dunal.) to NPK Fertilizers on the Nutrient Content and Uptake in Coastal Saline Soil,” vol. 13, pp. 955–957, Oct. 2013. Abstract
     A pot experiment was conducted at the pot culture yard of the Department of Soil Science and Agricultural Chemistry, Annamalai University with the objective to study the effect of different levels of NPK fertilizers on the nutrient content and uptake of Withania somnifera in coastal saline soil. The experiment was carried out in Completely Randomized Design with six treatments in four replications. The experiment consisted of application of NPK at different levels viz., T1 - Absolute control, T2 - 20:20:0 kg NPK ha-1, T3 - 30:30:10 kg NPK ha-1, T 4 - 40:40:20 kg NPK ha-1, T5 - 50:50:30 kg NPK ha-1 and T6 - 60:60:40 kg NPK ha-1. Ashwagandha variety Jawahar Asgandh-20 was grown as test crop. The results indicated that the increased rate of NPK fertilizer upto a level of 50:50:30 kg ha-1 significantly increased the N, P and K content and uptake of ashwagandha.
616. S. Venugopal, P. Saidaiah, and C. S. Karthik, “Chapter - 6 Genetics and Breeding of Ashwagandha (Withania Somnifera (L.) Dunal.),” p. 16.
617. S. Venugopal et al., “Comparative Investigation of the Genetic Components of Withania Somnifera L. Yield and Quality Traits Using Multivariate and Associations Analysis in the Deccan Plateau Region,” Industrial Crops and Products, vol. 188, p. 115696, Nov. 2022. doi: 10.1016/j.indcrop.2022.115696. Abstract
     The vital herb ashwagandha was farmed for commercial purposes in India. This study aimed to estimate the nature and amount of genetic variation in the 20 attributes in the 29 genotypes of ashwagandha in order to understand the correlations and character contributions to dry root weight/plant and alkaloid content with the selection of potential parents for ashwagandha crop improvement. Twenty-nine ashwagandha genotypes were assessed using a range of genetic metrics, including analysis of variance (ANOVA), genetic variability components, correlation, principal component analysis (PCA), and Mahalanobis D2. The ANOVA confirms a large amount of variability in the genotypes/varieties under study. Out of twenty the four traits, namely fresh leaf weight per plant (X8), dry leaf weight per plant (X9), seed yield per plant (X13), and dry root weight per plant (X16)—indicates the presence of additive gene action in the current study were highly heritable and had a high genetic advance mean value. Starch estimation, main root length, and root diameter were found to be positively and significantly correlated with dry root production per plant and total alkaloid content. The principle component analysis (PCA) results showed that the first four PCA accounted for the bulk (79.56%) of the differences. All genotypes are divided into four clusters based on the distances between and within clusters. Based on the findings mentioned above, it is recommended that genotypes from clusters IV (Poshita, CIMAP-Chetak), cluster II (AKAS-10, AKAS-11, NMITLI-101, and NMITLI-118), and cluster I (AKAS-02, IC-286632, and CIMAP Pratap) be chosen for a different hybridization program. Based on the multidisciplinary investigation, some of the genotypes with desired qualities have been found. They can be employed in upcoming breeding programs to increase alkaloid content and dry root yield.
618. S. Venugopal, M. Padma, M. Rajkumar, N. Seenivasan, P. Saidaiah, and G. Sathish, “Correlation and Path Analysis in Ashwagandha (Withania Somnifera L.) for Dry Root Yield,” Journal of Plant Development Sciences, vol. 13, no. 7, pp. 445–452, 2021. Abstract
     The experiment was laid out in a completely Randomized Block Design with 29 ashwagandha access ions as treatments during Kharif, 2018 at M edicinal and Aromatic Plant Research Station, Sri Konda Laxman Telangana State Horticultural University, Rajendranagar, Hyderabad. Each treatment was randomly replicated thrice. The results on genotypic and phenotypic correlation reveal that mostly genotypic correlation coefficient is comparatively higher than the intensity of phenotypic correlation coefficient. This indicates less influence of environment in association studies. The positive and significant correlation was observed between dry root yield per plant with root diameter, main root length, leaf length, starch estimation, leaf width, fresh leaf weight, dry leaf weight, days to flower initiation, plant height and number of secondary roots per plant. Direct selection based on these traits would result in simultaneous improvement of aforesaid traits and dry root yield per se in ashwagandha. Although correlation coefficients indicate the nature of association among the characters, path analysis splits the correlation coefficients into measures of direct and indirect effects, thus providing an understanding on the direct and indirect contribution of each character towards yield. From the foregoing discussion, it can be concluded that main root length, root diameter, leaf weight and days to flower initiation had positive correlation and positive direct effect on dry root yield per plant. These are identified as superior yield components. Hence, the genotypes which exhibited better performance for these characters can be used in further improvement of ashwagandha.
619. O. P. Verma, R. B. L. Gupta, and A. Shivpuri, “A New Host for Pithomyces Chartarum, the Cause of a Leaf Spot Disease on Withania Somnifera.,” Plant Pathology, vol. 57, no. 2, 2008. doi: 10.1111/j.1365-3059.2007.01732.x. Abstract
     This paper describes the first report of P. chartarum, the causal agent of a leaf spot, on W. somnifera, in India. Symptoms, morphology and pathogenicity tests confirmed the identity of the fungi.
620. S. K. Verma and A. Kumar, “Therapeutic Uses of Withania Somnifera (Ashwagandha) with a Note on Withanolides and Its Pharmacological Actions,” vol. 4, 2011. Abstract
     Withania somnifera is a medicinal plant extends over a large area, from the Atlantic ocean to South East Asia and from the Mediterranean region to South Africa. The medicinal plants are widely used by the traditional medical practitioners for curing various diseases in their day to day practice. In traditional systems of medicine, different parts (leaves, stem, flower, root, seeds, bark and even whole plant) of Withania somnifera (known as Ashwagandha in Hindi), a small herb seen throughout India, have been recommended for the treatment of aphrodisiac, liver tonic, antiinflammatory agent, astringent, and more recently to treat bronchitis, asthma, ulcers, emaciation, insomnia, and senile dementia etc. Clinical trials and animal research support the therapeutic use of ashwaganda for anxiety, cognitive and neurological disorders, inflammation, and Parkinson’s disease. Ashwaganda’s chemopreventive properties make it a potentially useful adjunct for the patients undergoing radiation and chemotherapy. Ashwaganda is also used therapeutically as an adaptogen for patients with nervous exhaustion, insomnia, and debility due to stress, and as an immune stimulant in patients with low white blood cell counts in blood. The major biochemical constituents of ashwaganda root are steroidal alkaloids and steroidal lactones in a class of constituents called withanolides.
621. G. Vijayabaskar and V. Elango, “Determination of Phytocompounds in Withania Somnifera and Smilax China Using GC-MS Technique,” Journal of Pharmacognosy and Phytochemistry, vol. 7, no. 6, pp. 554–557, 2018. https://www.phytojournal.com/archives/2018.v7.i6.6218/determination-of-phytocompounds-in-ltemgtwithania-somniferaltemgt-and-ltemgtsmilax-chinaltemgt-using-gc-ms-technique. Abstract
     GC-MS method used for the analysis of the obtained extract can be an interesting tool for testing the amount of some active principles in herbs used in various industries. The aim of this study was to carry out for identification of bioactive compounds from the whole plant methanolic extract of Withania somnifera and Smilax china by Gas chromatography and Mass spectroscopy (GC-MS). GCMS analysis of methanolic extract was done by standard protocol using the equipment Perkin-Elmer Gas Chromatography–Mass Spectrometry, while the mass spectra of the compounds found in the extract was matched with the National Institute of Standards and Technology (NIST) library. The GC-MS analysis revealed the presence of various compounds like Thirteen (13) compounds were identified in Withania somnifera whreas sixteen (16) compounds were identified in Smilax china. In the ethanolic extract of Withania somnifera and Smilax china these findings support the traditional use of Withania somnifera and Smilax china in various disorders.
622. S. Vinod, K. Natarajan, K. Santhanu, D. Selvakumar, S. Natesan, and K. Senthil, “Effect of Seaweed Elicitation and Culture Conditions on Biomass and Metabolite Production in in Vitro Shoots of Withania Somnifera,” South African Journal of Botany, vol. 151, pp. 781–790, Dec. 2022. doi: 10.1016/j.sajb.2022.10.051. Abstract
     Withania somnifera is an important medicinal plant used in various herbal preparations due the presence of its key metabolites-withanolides. The present study focuses on the effect of seaweed extract and different in vitro culture conditions on the biomass and key metabolite content in in vitro shoot cultures of Withania somnifera. The commercially available seaweed extract powder of Kappaphycus alverizii with the trade name ’Magic vita - P’ was examined for its effect on solid, suspension and bioreactor cultures of Withania somnifera. The standardisation of elicitation conditions showed the exposure of shoot cultures to 0.25% seaweed extract for 7 days resulted in maximum biomass and withaferin A content which were 1.91 fold and 3.05 fold higher than that of untreated shoots. Further among the three culture conditions studied, the bioreactor cultures exhibited maximum growth index compared to the solid and suspension cultures thus producing maximum biomass which in turn resulted in higher accumulation of withanolides per batch of culture. Thus the seaweed treated shoots in bioreactor yielded 18.56, 8.96, and 7.29 fold increase in withaferin A, withanolide A and withanone content compared to control shoots in solid media. Further the effect of this elicitation and culture conditions on the accumulation pattern of the primary metabolites were studied using GC-MS. The Principal Component Analysis (PCA) of the GC-MS data exhibited a clear distinction between the control and seaweed treated samples. Thus our findings indicate a clear protocol for scaling up of withanolide production from in vitro shoot cultures of Withania somnifera.
623. S. Vinotha, T. Ira, and S. Sri Ranjani, “Phytochemical Screening of Various Extracts of Root of Withania Somnifera (L.) Dunal,” 2013. http://repo.lib.jfn.ac.lk/ujrr/handle/123456789/6258. Abstract
     Withania somnifera (L) Dunal (Solanaceae), commonly known as Aswagandha, is one of the most valued medicinal plants with a number of pharmaceutical applications. Root extracts of W. somnifera are commonly used as a remedy for variety of ailments and a general tonic for overall health and longevity in the Traditional medicine system. It is also reported to have anti-inflammatory, anti-arthritic, antitumor, immunomoduatory and antioxidant effects. Although the phytochemical screening of W. somnifera has been already published, the aim of this study is to compile its phytochemical constituents in various extracts, which were carried out using standard laboratory procedures. Quantification of some of the active constituents like alkaloids, flavonoids and saponins were also carried out. The preliminary phytochemical screening of hot and cold ethanol, methanol and water extracts showed the presence of alkaloids, saponins, flavonoids, steroids, tannins, proteins, reducing sugar and coumarins and absence of quinones or anthraquinones. Cold and hot water extracts indicated the presence of fat and fixed oil. The total alkaloid and flavonoid content were found to be 0.81± 0.01% and 14.43±0.40% respectively, and total saponin content was (Foaming Index) FI < 100. The findings are consistent with the presence of biologically active constituents in the polar extracts of W. somnifera.
624. G. Vishal and S. D. B., “Ashwagandha (Withania Somnifera L. Dunal) Crop as Affected by the Application of Farm Yard Manure (FYM) and Inorganic Phosphorus in Typic Torripsamment of Hisar,” African Journal of Biotechnology, vol. 13, no. 6, pp. 743–748, Feb. 2014. doi: 10.5897/AJB2013.13538. Abstract
     The vegetative parameters of ashwagandha (viz. plant height, number of primary branches, plant spread and dry weight of shoot) were enhanced significantly with the application of 12.5 mg P2O5 kg-1 soil, whereas; dry weight of roots was enhanced up to the application level of 25 mg P2O5 kg-1 soil. FYM at the rate of 12.5 t ha-1 in combination with 12.5 mg P2O5 kg-1 soil significantly improved all the vegetative parameters, whereas, FYM at the rate of 12.5 t ha-1 in combination with 25 mg P2O5 kg-1 soil significantly enhanced the dry weight of the roots. The alkaloids yield (mg pot-1) in ashwagandha roots increased significantly with the application of 25 mg P2O5 kg-1 soil. The application of FYM at the rate of 12.5 t ha-1 improved the alkaloids yield (mg pot-1) but the significantly highest yield of total alkaloids (mg pot-1) was found in the treatment combination of 12.5 t FYM ha-1 + inorganic-P at the rate of 25 mg P2O5 kg-1 soil as compared with other eleven treatments. Nutrients (NPK) uptake by ashwagandha shoot increased significantly with the application level of 12.5 mg P2O5 kg-1 soil over control whereas in case of ashwagandha roots, the increase in nutrients uptake at the level of 25 mg P2O5 kg-1 soil over control. Application of FYM at the rate of 12.5 t ha-1 + 12.5 mg P2O5 kg-1 soil was the best treatment combination for ashwagandha roots in terms of nutrients uptake. Application of fertilizer-P significantly improved the status of organic carbon, Available-P in post harvest soil but decreased the Available-N, Available-K and DTPA extractable micronutrients (Fe, Cu, Zn and Mn) up to the level of 25 mg P2O5 kg-1 soil. FYM at the rate of 12.5 t ha-1 helped in maintaining the soil fertility status after harvest of the crop alone or in combination with fertilizer-P.
625. A. Vishnoi, S. B. Babbar, and S. C. Gupta, “Induction of Androgenesis in Anther Cultures of Withania Somnifera,” Zeitschrift für Pflanzenphysiologie, vol. 94, no. 2, pp. 169–171, Sep. 1979. doi: 10.1016/S0044-328X(79)80132-4. Abstract
     Callus formation has been induced in anthers of Withania somnifera cultured on Murashige and Skoog’s (1962) medium supplemented with BAP (10-6 M). The origin of the callus has been traced to pollen grains. The callus, further differentiated into embryoids.
626. G. Vitali, L. Conte, and M. Nicoletti, “Withanolide Composition and in Vitro Culture of Italian Withania Somnifera,” Planta Medica, vol. 62, no. 3, pp. 287–288, Jun. 1996. doi: 10.1055/s-2006-957884. Abstract
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627. R. Wadhwa, A. Konar, and S. C. Kaul, “Nootropic Potential of Ashwagandha Leaves: Beyond Traditional Root Extracts,” Neurochemistry International, vol. 95, pp. 109–118, May 2016. doi: 10.1016/j.neuint.2015.09.001. Abstract
     Rapidly increasing aging population and environmental stressors are the two main global concerns of the modern society. These have brought in light rapidly increasing incidence of a variety of pathological conditions including brain tumors, neurodegenerative & neuropsychiatric disorders, and new challenges for their treatment. The overlapping symptoms, complex etiology and lack of full understanding of the brain structure and function to-date further complicate these tasks. On the other hand, several herbal reagents with a long history of their use have been asserted to possess neurodifferentiation, neuroregenerative and neuroprotective potentials, and hence been recommended as supplement to enhance and maintain brain health and function. Although they have been claimed to function by holistic approach resulting in maintaining body homeostasis and brain health, there are not enough laboratory studies in support to these and mechanism(s) of such beneficial activities remain largely undefined. One such herb is Ashwagandha, also called "Queen of Ayurveda" for its popular use in Indian traditional home medicine because of its extensive benefits including anticancer, anti-stress and remedial potential for aging and neurodegenerative pathologies. However, active principles and underlying mechanism(s) of action remain largely unknown. Here we provide a review on the effects of Ashwagandha extracts and active principles, and underlying molecular mechanism(s) for brain pathologies. We highlight our findings on the nootropic potential of Ashwagandha leaves. The effects of Ashwagandha leaf extracts are multidimensional ranging from differentiation of neuroblastoma and glioma cells, reversal of Alzheimer and Parkinson’s pathologies, protection against environmental neurotoxins and enhancement of memory.
628. D. G. A. Walkey and J. M. G. Woolfitt, “Clonal Multiplication ofNicotiana Rustica L. from Shoot Meristems in Culture,” Nature, vol. 220, no. 5174, pp. 1346–1347, Dec. 1968. doi: 10.1038/2201346a0. Abstract
     WHEN meristem tips of the orchid, Cymbidium, are cultured, they do not give rise to one plant, but usually produce a mass of protocorms, each capable of developing into a plant1. Growth of meristems in shake culture increases the rate of multiplication of the protocorms2,3.
629. N. J. Walton, R. J. Robins, and M. J. C. Rhodes, “Perturbation of Alkaloid Production by Cadaverine in Hairy Root Cultures of Nicotiana Rustica,” Plant Science, vol. 54, no. 2, pp. 125–131, Jan. 1988. doi: 10.1016/0168-9452(88)90090-8. Abstract
     Cadaverine (1–10 mM) stimulated the production of anabasine by hairy root cultures of Nicotiana rustica transformed with Agrobacterium rhizogenes. In control cultures, nicotine accounted for at least 70–80% of the total alkaloid produced, whereas in cultures supplemented with 5 mM cadaverine about two-thirds of the alkaloid was anabasine and nicotine production was markedly diminished. Putrescine and agmatine caused some stimulation of alkaloid production, but the ratio of nicotine to anabasine was essentially unaffected. Lysine caused no substantial increase in anabasine formation.
630. L. Wang, B. Cheng, Z. Li, T. Liu, and J. Li, “Intelligent Tobacco Flue-Curing Method Based on Leaf Texture Feature Analysis,” Optik, vol. 150, pp. 117–130, Dec. 2017. doi: 10.1016/j.ijleo.2017.09.088. Abstract
     Most traditional curing systems are manually or half-artificial operated that requiring the curers to observe the state of tobacco leaves frequently. A novel intelligent real-time curing control system is developed in this paper by acquiring the optical image of tobacco leaves and extracting the color features and texture features to predict and control the temperature and humidity of the curing barn. The tobacco leaves changes from green to yellow and shrinks gradually, and this changing regulation would enhance the intelligence of tobacco curing system. The proposed neural network is designed to predict the set-point values of the adjustment of dry-bulb temperature, wet-bulb temperature and the changing time, which has eleven inputs include three color features, three texture features, ideal dry-wet temperature, ideal wet-bulb temperature, current stage, stage passing time, tobacco leaves varieties and flag. Some experiments are induced and the experimental results show this proposed approach based on color features and texture features could improve significantly the accuracy than that of the similar method only using color features especially in post-curing process.
631. S. G. Wankhade, S. V. Gholap, and P. P. Khode, “Effect of Seed Rates on the Root Yield and Quality of Ashwagandha (Withania Somnifera Dunal),” Agricultural Science Digest, vol. 29, no. 1, pp. 39–41, 2009. https://arccjournals.com/journal/agricultural-science-digest/.
632. A. Waratadar, P. J. Nirmalnath, P. S. Matiwade, A. Kr, and R. Ks, “Yield Parameter of Tobacco (Nicotiana Tabacum L.) as Influenced by AM Fungi under Orobanche Infested Soils,” Journal of Pharmacognosy and Phytochemistry, vol. 9, no. 1, pp. 301–304, 2020. https://www.phytojournal.com/archives/2020.v9.i1.10440/yield-parameter-of-tobacco-nicotiana-tabacum-l-as-influenced-by-am-fungi-under-orobanche-infested-soils. Abstract
     Tobacco is a one of the commercial products prepared from the tobacco leaves by curing them. Among the different species of tobacco, Nicotiana tabacum and Nicotiana rustica are well known tobacco species grown commercially across the world. Tobacco contains several phyto-chemicals like nicotine a principle alkaloid known for its insecticidal property in the form of nicotine. Recent studies have revealed that the AM fungal colonization is likely to induce growth promotional activities like number of leaves per plant and yield of tobacco per plot compared to uninoculated control under Orobanche infested soils. In this regard a field investigation was carried out to evaluate the three different methods of application of AM Fungal culture viz., planting of pre colonized tobacco seedling; soil application and the combination of both. The experiment was carried out in Orobanche infested soils of tobacco growing areas of Nipani in Belagavi district. The results of the present field investigation has revealed that the yield of tobacco at 150 DAP increased in the treatment received both planting of pre colonized tobacco seedling as well as soil application of STD AMF (2.52 kg/plot) which is significantly higher to the treatment received pre colonized tobacco seedling with UASDAMFT alone (2.44 kg /plot). However the lowest yield performance was recorded in uninoculated control tobacco plants (1.73kg/plot). Furthermore number of tobacco leaves per plant at 120 DAP increased in the treatment received pre colonized plus soil application of STD AMF at the time of planting documented the highest tobacco leaves (25.44 leaves/plant) compared to un inoculated control(17.55 leaves/plant). Thus our findings are of positive indicative of the effectiveness of application of AMF as mycorrhized seedlings followed by soil application will be a promising strategy to develop a growth promotional activity in tobacco under Orobanche infested soils.
633. N. G. Wasnik et al., “Establishment of in Vitro Root Cultures and Analysis of Secondary Metabolites in Indian Ginseng - Withania Somnifera,” Korean Journal of Plant Resources, vol. 22, no. 6, pp. 584–591, 2009. https://koreascience.kr/article/JAKO200916955022312.page. Abstract
     Adventitious root culture was established in the Jawahar variety of Withania somnifera using MS basal medium supplemented with 0.5 (mg/l) IAA and 2.0 (mg/l) IBA. Root tips from germinated seedlings, MS0 maintained plants and adventitious roots were maintained in suspension medium (1/2 MS basal medium supplemented with 3% sucrose) for a period of 1 to 6 months. The weight gain in roots was noted and the withanolides were extracted from the dry roots using solvents petroleum ether, 50% ethanol and chloroform. The withanolides in the chloroform fractions of all root samples analyzed were compared using thin layer chromatographic analysis. Withanolide content in adventitious root sample was found to be superior compared to other roots at any given point of time during the 6month growth period.HPLC analysis of in vitro adventitious roots showed the presence of a new compound.
634. D. Weckenmann and P. Martin, “Endopeptidase Activity and Nitrogen Mobilization in Senescing Leaves of Nicotiana Rustica in Light and Dark,” Physiologia Plantarum, vol. 60, no. 3, pp. 333–340, 1984. doi: 10.1111/j.1399-3054.1984.tb06072.x. Abstract
     The time course of endopeptidase activity (digestion of azocasein at pH 4.6) in leaves of intact plants of Nicotiana rustica L. was studied and related to changes in the contents of chlorophyll, total nitrogen and soluble and insoluble protein nitrogen. Endopeptidase activity increased several fold during senescence. However, the course of protein degradation did not reflect the steep slope of azocaseolytic activity. When single mature leaves were darkened, senescence proceeded faster than in illuminated leaves but the amount of nitrogen mobilized and translocated did not differ greatly between darkened and illuminated leaves. However, in contrast to leaves in light, azocaseolytic activity did not increase. Gelatin zymograms obtained using isoelectric focusing of extracts of mature leaves showed several bands in the pH 4.0 to 6.5 region of the gels. During senescence in both light and dark the position and number of bands remained largely unchanged. In leaves in light, the activity of endopeptidases focusing in the range pH 4.1 to 5.0 increased greatly. In leaves in dark, however, no major changes in activity could be detected. The results suggest that in tobacco leaves endopeptidase activity normally increases considerably during senescence but this increase is not a prerequisite for an effective protein degradation. Separation and analysis of free amino acids showed that during senescence in light the levels of all amino acids decreased considerably. In leaves senescing in the dark there were large increases in the levels of glutamine and asparagine, concomitant decreases in glutamate and aspartate, and considerable increases in all other amino acids.
635. R. Wellington, “Studies of Natural and Artifical Parthenogenesis in the Genus Nicotiana,” The American Naturalist, vol. 47, no. 557, pp. 279–306, May 1913. doi: 10.1086/279349. Abstract
     1. Seed giving plants true to the maternal species in the F1 generation accompanied by aborted seed probably hybrid in nature, was found when certain Nicotiana species were cross-fertilized. Hybrid plants and plants purely maternal were obtained from the same capsules in other crosses. 2. The capsules of several Nicotiana species were caused to swell slightly by merely tickling them with a sharp-pointed instrument, but no seeds were produced. 3. Abortive seed probably without embryos was produced by singeing young buds with a hot platinum wire, by the exposure of young plants to chloroform gas, and by cutting away a portion of the pistil and pollinating the stub both with and without the accompaniment of a germinative fluid. 4. Abortive seed was produced by shortening the pistils of a flower and grafting the stigma end of another pistil on to the stub and pollinating the same. 5. The ringing of the branches below a cluster of buds did not assist in the production of seed. 6. No seed was produced by the simple methods of emasculation and decapitation of blossoms, except in one doubtful case of N. phumbaginifolia. 7. It is likely that an agent inhibitory to the growth of pollen grains is present in the stigmatic fluids of certain species of the genus Nicotiana; at least, the pollen grains of N. suaveolens did not germinate in N. Forgetiana stigmatic fluid when placed within a Van Tieghem cell. 8. The exposure of young N. rustica var. texana, plants to acetone gas caused the transformation Of the corollas and the stamens of most of the terminal flowers into leafy tissue; otherwise, except in the mentioned case of the chloroform, no results were secured by the use of anaesthetic and toxic gases. 9. The injection of chemicals into the stems of tobacco plants was valueless in the production of seed. 10. As no unquestionable case of parthenogenetic seed was produced in the several hundred trials, it seems very improbable that parthenogenesis exists in the genus Nicotiana-at least in the species tested. The seed obtained in the crosses which came true to the mother species is probably polyembryonic-the stimulus of development being imparted either by the penetrating pollen tubes or by a substance exuded from the same.
636. M. WenGuang et al., “Effects of seed priming with different agents on seed germination and seedling growth in tobacco (Nicotiana tabacum L.) under drought stress.,” Acta Agriculturae Zhejiangensis, vol. 24, no. 6, pp. 949–956, 2012. https://www.cabdirect.org/cabdirect/abstract/20133031151. Abstract
     The effects of different agents on seed germination and growth characteristics of tobacco varieties with different drought tolerance were studied in this paper. Seeds of two tobacco varieties with different drought tolerance were primed in different concentration solutions of paclobutrazol, calcium chloride, proline and salicylic acid. Then, the germination, seedling growth and physiological...
637. C. P. Werner, I. M. Dunkin, M. A. Cornish, and G. H. Jones, “Gene Transfer in Nicotiana Rustica by Means of Irradiated Pollen II Cytogenetical Consequences,” Heredity, vol. 52, no. 1, pp. 113–119, Feb. 1984. doi: 10.1038/hdy.1984.11. Abstract
     Pollen from Nicotiana paniculata and the V12 variety of N. rustica was irradiated with a range of high doses of γ-rays up to 100 Krads. Both kinds of pollen were used to pollinate the V27 variety of N. rustica. Radiation treatments above 30 Krads gave no viable seed. A cytological examination of the M1 progeny from the 20 Krad treatments of both crosses revealed conventional radiation damage in the form of losses of whole chromosomes and parts of chromosomes, and rearrangements. The plants possessed hybrid or aberrantly hybrid phenotypes. It was concluded that they were the products of a conventional fertilisation mechanism rather than the gene transfer mechanism proposed by Pandey (1980). The expression of mutational damage can probably account for most of the maternal trends observed in the intervarietal M2 of N. rustica examined previously, although post-meiotic selection may also play a role.
638. L. A. White, “Nicotiana Rustica Cultivated by Pueblo Indians,” Science, Jul. 1941. https://www.science.org/doi/abs/10.1126/science.94.2429.64-b.
639. J. J. Woodend and E. Mudzengerere, “Inheritance of Resistance to Wildfire and Angular Leaf Spot Derived from Nicotiana Rustica Var. Brasilea,” Euphytica, vol. 64, no. 1, pp. 149–156, Jan. 1992. doi: 10.1007/BF00023549. Abstract
     The introgression of wildfire (races 0 and 1) and angular leaf spot (ALS) resistance from N. rustica var. Brasilea into N. tabacum has proved economically useful in Zimbabwe although the mode of inheritance of, and genetic relationships between the resistance are unknown. This study was undertaken to (1) examine the mode of inheritance of the resistance to races 0 and 1 of wildfire, and ALS, (2) determine the genetic relationship between the resistances and (3) establish whether the N. rustica-derived wildfire race 0 resistance is allelic to that obtained from N. longiflora. Inheritance was examined under greenhouse and field conditions by studying disease reactions in the parental, F1, F2 and backcross generations derived from crosses of three susceptible lines to a resistant line Nr-7. Three-point backcrosses to the susceptible parent were examined for linkage and segregating generations from a cross of Nr-7 to Burley 21 which carries the N. longiflora race 0 resistance were used to test for allelism. In general, we observed that all resistances are determined by a single dominant gene although some incosistent ratios were obtained likely due to misclassification of disease reactions and erratic transmission. All resistances showed linkage although pleiotropism cannot be ruled out. Allelism tests demonstrated that the N. rustica race 0 resistance is not allelic to that obtained from N. longiflora. Our findings are examined in relation to the efficacy of indirect selection for resistance.
640. S. C. Wooley and T. D. Paine, “Infection by Mycorrhizal Fungi Increases Natural Enemy Abundance on Tobacco (Nicotiana Rustica),” Environmental Entomology, vol. 40, no. 1, pp. 36–41, Feb. 2011. doi: 10.1603/EN10145. Abstract
     The presence of arbuscular mycorrhizal fungi (AMF) influences plant nutrient uptake, growth, and plant defensive chemistry, thereby directly influencing multi-trophic interactions. Different fungal isolates (genotypes of the same fungal species) have been shown to differ in nutrient uptake ability. Plants infected with different AMF genotypes may vary in foliar nutrient or defensive chemical levels, potentially influencing multi-trophic interactions. Using a completely randomized design, we compared the effect of two isolates of the mycorrhizal fungus Glomus etunicatum W. N. Becker & Gerdemann on silver leaf whitefly (Bemisia argentifolii Bellows & Perring) (Hemiptera: Aleyrodidae) and parasitic wasp (Eretmocerus eremicus Rose & Zolnerowich) (Hymenoptera: Aphelinidae) abundance. Whitefly populations were not influenced by AMF infection. Parasite populations were higher on plants infected with the isolate collected from Georgia, even after controlling for whitefly abundance and plant architecture. We propose that AMF indirectly influences parasite abundance and parasitism through a change in leaf surface chemicals that affect parasitic wasps. Because of the ubiquity of and genetic variation in AMF, multi-trophic interactions are likely to be strongly influenced by belowground processes.
641. G. Woras, E. A. Hashmi, F. Karim, M. Shahid, and S. A. Shah, “Comparative Evaluation of Some Tobacco Varieties (Nicotiana rusticaL.),” Pak Tobacco (Pakistan), 2004. https://agris.fao.org/search/en/records/6472434d53aa8c896303e74e. Abstract
     Rustica tobacco varieties (Nicotiana rustica L.) were evaluated foryield and yield components at the Tobacco Research Station, Mardanduring the year 1997-98. The experiment was laid out according to ,Randomized Complete Block Design replicated thrice. The results revealedthat varieties were significantly different for all four characters studied.Variety Rustica-14 had maximum plant height. Maximum leaf size, numberof leaves/plant and cured leaf yield were exhibited by variety Rustica-13followed by variety Rustica-14. On the basis of better performance,varieties Rustica-13 and 14 are recommended for Mardan area.
642. Y. N. Xin, J. W. Zhang, and B. Li, “Drying Kinetics of Tobacco Strips at Different Air Temperatures and Relative Humidities,” Journal of Thermal Analysis and Calorimetry, vol. 132, no. 2, pp. 1347–1358, May 2018. doi: 10.1007/s10973-018-7005-5. Abstract
     The effects of air temperature and relative humidity (RH) on the drying kinetics of two kinds of flue-cured tobacco strips (C3F and B3F) were studied using a thermogravimetric device in this work. The drying experiments were carried out with drying air temperatures of 60, 70, 80, and 90 °C and RH values of 0, 10, 20, 30, and 40% at a constant airflow. Taking the effect of RH into consideration, a modified Arrhenius-type equation of diffusivity was proposed. In addition, comparing five thin-layer drying models and five equilibrium moisture content models to describe the drying kinetics and the desorption isotherms of tobacco strips, high coefficients of determination (R2) and low reduced Chi-square (χ2) and residual sum of squares (RSS) values indicated that the Logarithmic model and the Modified Oswin model appeared to be the most suitable for predicting the moisture ratio and the moisture desorption relationship of tobacco strips. The effective moisture diffusion coefficient under different temperatures and RHs ranged from 1.68 × 10−11 to 6.81 × 10−11 m2 s−1 for C3F and from 1.62 × 10−11 to 6.68 × 10−11 m2 s−1 for B3F. A modified Arrhenius-type equation with an RH term was found to be effective for describing the drying behavior of two flue-cured tobacco strips, and the activation energy (Ea) values were 34.6 and 35.2 kJ mol−1 for C3F and B3F, respectively.
643. G. Xu et al., “Transcriptomic Insights into the Regulatory Networks of Chilling-Induced Early Flower in Tobacco (Nicotiana Tabacum L.),” Journal of Plant Interactions, vol. 17, no. 1, pp. 496–506, Dec. 2022. doi: 10.1080/17429145.2022.2055175. Abstract
     Appropriate timing of flowering is pivotal for tobacco, while chilling stress occurring at the seedling stage often undesirably leads to early flowering. However, the potential mechanism underlying chilling-induced early flowering remains unknown. Here, transcriptome sequencing was performed in tobacco with or without chilling at both seedling and budding stages. Chilling affected the expression of numerous genes at the seedling stage, while these dramatic expression changes were largely eliminated at the budding stage. A small number of genes related to metabolism, flower development, and stress tolerance continued to keep their altered expression patterns from the seedling stage to the budding stage. Many potential flowering-related genes involved in flowering pathways were identified and over half of them were differentially expressed. Functional analysis revealed that the down-regulation of NbXTH22 rendered tobacco less sensitive to chilling-induced early flowering. These results provide valuable resources for the investigation of flowering regulatory mechanisms and contribute to the genetic improvement of crops.
644. T. Yamada, W. Marubashi, and M. Niwa, “Facile Induction of Apoptosis into Plant Cells Associated with Temperature-Sensitive Lethality Shown on Interspecific Hybrid from the Cross Nicotiana Suaveolens × N. Tabacum,” Plant and Cell Physiology, vol. 42, no. 2, pp. 204–213, Feb. 2001. doi: 10.1093/pcp/pce029. Abstract
     Two lines of suspension culture cells were obtained from a hybrid seedling of Nicotiana suaveolens Lehm. × N. tabacum L. cv. Hicks-2 expressing temperature-sensitive lethality. One of them (LH line) was inducible cell death in accordance with the lethality at 28°C but not under high-temperature conditions (36°C). Another one (SH line) lost the lethality and survived at 28°C. The cells of LH line showed apoptotic changes when they were cultured at 28°C. Fragmentation of nuclei was correlated with the lethality in the cells, as confirmed by fluorimetry of the nuclear DNA using laser scanning cytometry. Agarose gel analysis of DNA extracted from the cells expressing the lethality revealed a specific ladder pattern suggesting nucleosomal fragmentation that is one of the biochemical characteristics of apoptosis. From these facts, we confirmed that the process of cell death leading to hybrid lethality in the cells is certainly apoptosis. Hybrid cells were used in the experiments to estimate the point of no return in temperature-sensitive lethality and to examine the influence of cation in DNA fragmentation during apoptosis. The utility of hybrid cells as an experimental system for studies of hybrid lethality and apoptosis in plants was confirmed.
645. N. Yamaguchi, S. Suzuki, and A. Makino, “Starch Degradation by Alpha-Amylase in Tobacco Leaves during the Curing Process,” Soil Science and Plant Nutrition, vol. 59, no. 6, pp. 904–911, Dec. 2013. doi: 10.1080/00380768.2013.842884. Abstract
     Tobacco (Nicotiana tabacum L.) leaf starch is degraded to sugars through curing (42°C/82.3% relative humidity/72 h). Total carbohydrate content remained almost constant, starch content decreased markedly, and soluble sugar content (mostly glucose) increased. α-Amylase and starch phosphorylase activities increased sixfold and threefold, respectively, whereas β-amylase activity was unaltered and isoamylase activity decreased. Increased α-amylase activity was accompanied by increased α-amylase protein levels. Although tobacco has four α-amylase gene members, only NtAMY1 mRNA levels increased. For other starch degradation genes, such as NtBAM1 and NtBMY2 (β-amylase), NtISO1 and NtISO2 (isoamylase) and NtGWD1 and NtGWD3 (glucan water dikinase), the mRNA levels remained unaltered during the first 48 h of curing. NtAMY1 expression was induced by osmotic stress but was unaffected by high temperature and/or injury stresses. Similarly, soluble sugar contents were largely increased by osmotic stress. This suggests that starch is degraded by α-amylase during curing and that α-amylase is coded by NtAMY1, induced by osmotic stress.
646. A. Yarden and D. Lavie, “567. Constituents of Withania Somnifera. Part I. The Functional Groups of Withaferin,” Journal of the Chemical Society (Resumed), no. INVALID_SCITE_VALUE, pp. 2925–2927, Jan. 1962. doi: 10.1039/JR9620002925. Abstract
     No abstract available
647. Y. O. Ya., “Use of Macroelements by Belladonna Plants by Stages of Development,” European Journal of Technical and Natural Sciences, no. 2-3, 2021. doi: 10.29013/EJTNS-21-2.3-9-15. Abstract
     The need for research of technologies for growing Belladonna vulgaris as a medicinal raw material is justified. The peculiarities of Belladonna plant nutrition at different stages of development are studied. The use of microelements depending on the physiological needs of plants is analyzed. The dynamics of changes in mineral nutrition compounds in plant tissues depending on the stages of organogenesis is given. It was found that the concentration of compounds in plant tissues varied and reached its maximum values before the generative stage of organogenesis. The obtained results show the dynamics of the concentration of mineral nutrition compounds in the dry matter of different parts of Belladonna plants in the process of their growing season. Accordingly, the need for mineral nutrition compounds depended on the stages of organogenesis that plants underwent.
648. N. YongZhi et al., “Research progress on the regulation mechanism of pollen-pistil interactions in Nicotiana.,” Journal of Yunnan University - Natural Sciences Edition, vol. 40, no. 6, pp. 1254–1260, 2018. https://www.cabdirect.org/cabdirect/abstract/20193098097. Abstract
     Pollination is a key process in life cycle of plants. The plants of Solanaceae have solid styles and wet stigmas. Successful pollination begins with pollen adhering to the surface of the mature stigma, followed by hydration and germination. The pollen tube passes through the stigma and style, targeting to the embryo sac inside the ovule, and the sperm is released to complete double fertilization...
649. C. Zaghdoud et al., “Growth and Nitrogen Metabolism Changes in NaCl-Stressed Tobacco (Nicotiana Rustica L. Var. Souffi) Seedlings,” African Journal of Biotechnology, vol. 12, no. 12, 2013. doi: 10.4314/ajb.v12i12. Abstract
     While salinity effects have been largely documented in crop plants, little data are available on Nicotiana rustica species (snuff tobacco), mainly nitrogen metabolism changes. Here, tobacco (N. rustica L. var. Souffi) seedlings were grown for one month on control medium, and then exposed for seven days to different levels of salt stress (0, 50, 100 and 200 mM NaCl). A significant decrease in dry weight accumulation was observed only at 200 mM NaCl. Na+ and Cl- ions accumulation was greater in leaves relative to roots, while growth was similarly decreased in both organs. Referring to roots, leaf water content was less affected by salinity. The increase of endogenous levels of salt ions was accompanied by a drop in NO3- content in both leaves and roots, and a more pronounced decrease in K+ content in leaves. Under salinity, nitrate reductase activity (NR, EC 1.6.1.6) was inhibited in both leaves and roots. The activities of nitrite reductase (NiR, EC 1.7.7.1) and glutamine synthetase (GS, EC 6.3.1.2) were inhibited in leaves but not in roots by salt stress. In stressed seedlings, NH4+ contents, protease activity, aminating and deaminating activities of glutamate dehydrogenase (NADH-GDH and NAD-GDH, EC 1.4.1.2) were enhanced mainly in the leaves. It could be stated that tobacco leaves and roots exhibited similar salt sensitivity in terms of growth and NO3- assimilation (NR activity), however they showed differential response for salt ions accumulation and NH4+ metabolism steps (GS and GDH).Keywords: Assimilation, growth, salt stress, nitrogen, mineral nutrition, tobaccoAfrican Journal of Biotechnology Vol. 12(12), pp. 1392-1400
650. M. Zaim and A. Samad, “Association of Phytoplasmas with a Witches-Broom Disease of Withania Somnifera (L.) Dunal in India,” Plant Science, vol. 109, no. 2, pp. 225–229, Aug. 1995. doi: 10.1016/0168-9452(95)04158-Q. Abstract
     A witches-broom disease of Withania somnifera first appeared in 1988 at CIMAP experimental fields, Lucknow, India. Thereafter, it spread to the commerical fields causing severe damage to the crop during the last 5 years. Phytoplasma was associated with the disease on the basis of symptomatology, mode of transmission, electron-microscopic studies and response of tetracycline spray on the recovery from diseased symptoms. Typical symptoms of the disease consisted of little leaf, shortening of internodes, excessive branching giving witches-broom appearance and premature drying and death of infected twigs and leaves. Ultrathin sections of infected leaf tissue showed pleomorphic bodies (measuring 250–550 nm) in sieve tubes of phloem. Disease was transmitted from infected plants to healthy plants through grafting. The treatment of infected plants with tetracycline provided temporary recovery from the infection.
651. J. Zeng et al., “Engineering the Production of Medicinal Tropane Alkaloids through Enhancement of Tropinone and Littorine Biosynthesis in Root Cultures of Atropa Belladonna,” Industrial Crops and Products, vol. 189, p. 115778, Dec. 2022. doi: 10.1016/j.indcrop.2022.115778. Abstract
     Hyoscyamine and scopolamine are two pharmaceutical tropane alkaloids (TAs) whose biosynthesis is dependent on tropinone and littorine. The biosynthesis process of tropinone and littorine was elucidated only very recently, with the discoveries of four novel TAs biosynthesis genes, namely type III polyketide synthase (PYKS), tropinone synthase (CYP82M3), phenyllactate UDP-glycosyltransferase (UGT1), and littorine synthase (LS). Yet their values in engineering TAs production remain unknown. In this study, their roles in engineering TAs’ production were studied via gene overexpression methods applied to root cultures of Atropa belladonna. PYKS or CYP82M3 overexpressed separately did not or slightly affected the biosynthesis of tropine, and had no effect on the accumulation of TAs. Co-overexpression of PYKS and CYP82M3 led to much higher levels of tropinone and tropine than those produced by the overexpression of either PYKS or CYP82M3. Overexpression of either UGT1 or LS significantly elevated the production of littorine, hyoscyamine, anisodamine, and scopolamine. Higher levels of these TAs were produced when UGT1 and LS were overexpressed together. Finally, when the four TA biosynthesis genes were overexpressed together, the highest production of TAs was obtained, indicating that synergistic effect did significantly augment TAs’ production.
652. F. F. Zenkner, M. Margis-Pinheiro, and A. Cagliari, “Nicotine Biosynthesis in Nicotiana: A Metabolic Overview,” Tobacco Science, vol. 56, no. 1, pp. 1–9, Apr. 2019. doi: 10.3381/18-063. Abstract
     Alkaloids are important compounds found in Nicotiana plants, essential in plant defense against herbivores. The main alkaloid of Nicotiana tabacum, nicotine, is produced in roots and translocated to the leaves. Nicotine is formed by a pyrrolidine and a pyridine ring in a process involving several enzymes. The pyridine ring of nicotine is derived from nicotinic acid, whereas the pyrrolidine ring originates from polyamine putrescine metabolism. After synthesis in root cortical cells, a set of transporters is known to transport nicotine upward to the aerial part and store it in leaf vacuoles. Moreover, nicotine can be metabolized in leaves, giving rise to nornicotine through the N-demethylation process. Some Nicotiana wild species produce acyltransferase enzymes, which allow the plant to make N-acyl-nornicotine, an alkaloid with more potent insecticidal properties than nicotine. However, although we can find a wealth of information about the alkaloid production in Nicotiana spp., our understanding about nicotine biosynthesis, transport, and metabolism is still incomplete. This review will summarize these pathways on the basis on recent literature, as well as highlighting questions that need further investigation.
653. B. Zhang, X.-qin Wang, X. Li, Y.-qing Ni, and H.-yu Li, “Aluminum Uptake and Disease Resistance in Nicotiana Rustica Leaves,” Ecotoxicology and Environmental Safety, vol. 73, no. 4, pp. 655–663, May 2010. doi: 10.1016/j.ecoenv.2009.12.028. Abstract
     The comparative effectiveness of aluminum hydroxide and aluminum chloride has been studied in the development of bacterial wilt infection on leaves of Nicotiana rustica cv. Gansu yellow flower. We have analyzed the changes of foliar H2O2 content, as well as of non-enzymatic and enzymatic antioxidants under aluminum stress. Pretreatment with aluminum hydroxide before pathogen challenge reduced the development of Ralstonia solanacearum infection and decreased the extent of leaf injury. The pretreatment also reduced the Al uptake in comparison to pretreatment with aluminum chloride. H2O2 generation was significantly enhanced by pretreatment with aluminum hydroxide. Increased NADPH oxidase and superoxide dismutase activities were correlated with limited infection. Aluminum hydroxide pretreatment shifted the leaf redox homeostasis of AsA/DHA and GSH/GSSG toward oxidation, yielding higher oxidant levels than aluminum chloride before bacterial inoculation. The results support the idea that aluminum hydroxide induced H2O2 accumulation through non-enzymatic and enzymatic regulation, ultimately resulting in resistance to tobacco wilt disease.
654. C.-S. Zhang, Y. Zheng, L. Peng, and J. Cao, “Rootstock-Scion Interaction Affects the Composition and Pathogen Inhibitory Activity of Tobacco (Nicotiana Tabacum L.) Root Exudates,” Plants, vol. 9, no. 12, p. 1652, Dec. 2020. doi: 10.3390/plants9121652. Abstract
     The composition and allelopathy to Phytophthora nicotianae (the causal agent of tobacco black shank disease) of root exudates from a resistant tobacco (Nicotiana tabacum L.) cultivar Gexin 3, a susceptible cultivar Xiaohuangjin 1025 and their reciprocal grafts were investigated. Grafting with disease-resistant rootstock could improve resistance to black shank; this is closely related to the allelopathy of root exudates. The root exudates from the resistant cultivar inhibited the growth of P. nicotianae, while those from the susceptible cultivar promoted the growth; the grafting varieties had intermediate properties. The root exudate composition differed among cultivars. Gexin 3 was rich in esters and fatty acids, while Xiaohuangjin 1025 contained more hydrocarbons and phenolic acids. The composition of root exudates of grafted cultivars as well as their allelopathy to P. nicotianae were altered, and tended to be close to the composition of cultivar used as rootstock. Eugenol, 4-tert-butylphenol, mono (2-ethylhexyl) phthalate, 4-hydroxybenzoic acid, 2,6-di-tert-butylphenol, dipropyl phthalate, and methyl myristate were identified as the main compounds contributing to inhibitory properties of root exudates. Sorbitol was suggested to play a role in disease induction. Overall, rootstock–scion interaction affected the composition of tobacco root exudates, which may be attributed to the different disease resistance among grafted plants, rootstock and scion.
655. S. Zhao, Z. Wu, M. Lai, M. Zhao, and B. Lin, “Determination of Optimum Humidity for Air-Curing of Cigar Tobacco Leaves during the Browning Period,” Industrial Crops and Products, vol. 183, p. 114939, Sep. 2022. doi: 10.1016/j.indcrop.2022.114939. Abstract
     Hainan is a natural air-curing barn for cigar leaves. Humidity has a marked influence on the air-curing of cigar leaves. The browning stage is a key period, which determines the degree and quality of cigar tobacco leaf browning. To understand the effects of humidity in the browning stage on tobacco leaf quality, we examined the malondialdehyde and polyphenol contents, polyphenol oxidase activity, moisture, chemical composition, and appearance of tobacco leaves that were air cured at humidities of 60%, 70%, and 80% during the browning stage. High humidity (80%) during curing led to a high moisture content and polyphenol oxidase activity in leaves. With regard to malondialdehyde content, a minute differences between moderate-humidity (70%) and high-humidity (80%) air-curing were observed. During the browning stage, the chlorogenic acid content was reduced by 85.32% when tobacco leaves were air-cured at 80% humidity, which means that it underwent a more comprehensive oxidized process compared to the other two treatments, whereas rutin and scopoletin experienced a more thorough oxidation at 60% humidity. 70% Humidity was found to be the most suitable humidity for the appearance of air-cured leaves than others, producing the optimum appearance of the cigar leaves. At the end of air-curing, the treatment with 70% curing humidity had a more coordinated chemical composition compared to the other treatments, for instance, it had a starch content of 1.14% and a potassium-chloride ratio of 7.65, which indicated a much better quality of cigar tobacco leaves. In summary, these findings indicate that air-curing at 70% humidity during the browning stage was the most favorable choice for improving tobacco leaf quality for cigar production.
656. B. Zhao, F. A. Agblevor, R. K. C., and J. G. Jelesko, “Enhanced Production of the Alkaloid Nicotine in Hairy Root Cultures of Nicotiana Tabacum L.,” Plant Cell, Tissue and Organ Culture (PCTOC), vol. 113, no. 1, pp. 121–129, Apr. 2013. doi: 10.1007/s11240-012-0256-0. Abstract
     The utility of hairy root cultures to produce valuable phytochemicals could be improved by repartitioning more of the desired phytochemical into the spent culture media, thereby simplifying the bioprocess engineering associated with the purification of the desired phytochemical. The majority of nicotine produced by tobacco hairy root cultures is retained within roots, with lesser amounts exuded into the spent culture media. Reduced expression of the tobacco nicotine uptake permease (NUP1) results in significantly more nicotine accumulating in the media. Thus, NUP1-reduced expression lines provide a genetic means to repartition more nicotine into the culture media. The present study examined a wild type and a NUP1-reduced expression hairy root line during a variety of treatments to identify culture conditions that increased nicotine accumulation in the media. The NUP1-reduced expression line grew faster, used less oxygen, and exuded more nicotine into the media. Basification of the culture media associated with root growth resulted in a dramatic reduction in nicotine accumulation levels in the media, which was reversed by decreasing the pH of the media. Kinetic analysis of hairy root growth and nicotine accumulation in the media revealed a potential improvement in nicotine yields in the media by stimulating the branching of tobacco hairy roots.
657. Z.-L. Zheng, Z. Yang, J.-C. Jang, and J. D. Metzger, “Phytochromes A1 and B1 Have Distinct Functions in the Photoperiodic Control of Flowering in the Obligate Long-Day Plant Nicotiana Sylvestris,” Plant, Cell & Environment, vol. 29, no. 9, pp. 1673–1685, 2006. doi: 10.1111/j.1365-3040.2006.01538.x. Abstract
     The obligate long-day plant Nicotiana sylvestris with a nominal critical day length of 12 h was used to dissect the roles of two major phytochromes (phyA1 and phyB1) in the photoperiodic control of flowering using transgenic plants under-expressing PHYA1 (SUA2), over-expressing PHYB1 (SOB36), or cosuppressing the PHYB1 gene (SCB35). When tungsten filament lamps were used to extend an 8 h main photoperiod, SCB35 and SOB36 flowered earlier and later, respectively, than wild-type plants, while flowering was greatly delayed in SUA2. These results are consistent with those obtained with other long-day plants in that phyB has a negative role in the control of flowering, while phyA is required for sensing day-length extensions. However, evidence was obtained for a positive role for PHYB1 in the control of flowering. Firstly, transgenic plants under-expressing both PHYA1 and PHYB1 exhibited extreme insensitivity to day-length extensions. Secondly, flowering in SCB35 was completely repressed under 8 h extensions with far-red-deficient light from fluorescent lamps. This indicates that the dual requirement for both far-red and red for maximum floral induction is mediated by an interaction between phyA1 and phyB1. In addition, a diurnal periodicity to the sensitivity of both negative and positive light signals was observed. This is consistent with existing models in which photoperiodic time measurement is not based on the actual measurement of the duration of either the light or dark period, but rather the coincidence of endogenous rhythms of sensitivity – both positive and negative – and the presence of light cues.
658. P. Zhou et al., “Transcriptomic Analyses of Chilling Stress Responsiveness in Leaves of Tobacco (Nicotiana Tabacum) Seedlings,” Plant Molecular Biology Reporter, vol. 38, no. 1, pp. 1–13, Mar. 2020. doi: 10.1007/s11105-019-01167-0. Abstract
     Low temperature is among the most significant abiotic stresses restricting geographical distribution of plants and reducing crop productivity. However, the molecular regulatory mechanisms of tobacco plants in response to low temperature are poorly understood. To elucidate the molecular mechanisms of chilling tolerance in tobacco, the transcriptomic responses of tobacco under chilling stress were analyzed using RNA-seq analysis. A total of 1675 differentially expressed genes (DEGs) were detected from T12h vs. CK12h and T24h vs. CK24h libraries; among these genes, 1170 genes were upregulated and 505 were downregulated. Additionally, 109 genes were found to be specifically expressed in tobacco seedlings under chilling stress. Functional annotation revealed that the DEGs enriched that categories of regulating soluble sugar and polyamine content and composition to maintain cell osmotic potential, accelerating the de novo synthesis of D1 protein to promote PSII repair, regulating signal transduction such as ABA and GA, and promoting lipid metabolism and lignin synthesis to improve stability of membrane system and mechanical strength of cell wall. This work provides additional insights into the molecular basis of tobacco seedling responses to low-temperature stress.
659. M. K. Zubko, E. I. Zubko, O. A. Khvedynich, S. V. Lopato, S. A. Latipov, and \relax Y. \relax Y. Gleba, “Somatic Hybridization Between Nicotiana Tabacum L. (Tobacco) and Atropa Belladonna L. (Deadly Nightshade),” in Somatic Hybridization in Crop Improvement II, T. Nagata and Y. P. S. Bajaj, Eds. Berlin, Heidelberg: Springer, 2001, pp. 304–327. doi: 10.1007/978-3-642-56758-2_21. Abstract
     Somatic hybridization based on protoplast fusion appeared as an alternative to sexual hybridization of higher plants. The term was proposed by (1974). Numerous experiments on somatic hybridization have been carried out using Nicotiana species (Gleba and Sytnik 1984; Negrutiu et al. 1989; Bates 1992; Puite 1992). The genus Nicotiana belongs to the Solanaceae family and includes about 70 species showing significant genetic and morphological variability (Goodspeed 1954). The most important species is N. tabacum L., or tobacco, cultivated widely as a source for the tobacco industry. N. tabacum (2n=48) originated from Central and South America (Darlington and Wylie 1955) but is now unknown in the wild. Different varieties of tobacco display strong vigor of stems possessing simply organized large leaves covered by abundant trichomes. Determinate inflorescences contain about 60–100 pink flowers, which after self- or cross-pollination develop capsules full of small brown seeds. Due to these characteristics, which allow easy propagation and manipulation with cells and tissues in vitro, tobacco became a convenient model species for plant cell- and genetic engineering. The success of wholeplant regeneration from cultivated tobacco protoplasts (Takebe et al. 1971) further provided basis for the generation of hybrids by protoplast fusions. First somatic hybrids were produced in interspecific Nicotiana combinations (Carlson et al. 1972; Melchers and Labib 1974; Gleba et al. 1975; Kameya 1975).