Nicotiana rustica

https://tobacco.ces.ncsu.edu/

Propagation

Germination

[1] [2] [3] [4] [5] [6] [7] [8] [9] [10] [11] [12] [13] [14] [15] [16] [17] [18] [19] [20] [21]

Seeds can be surface sterilized using 5% sodium hypochlorite.[22]

40-day transplant time.[23]

Vegetative

In-Vitro

Cultivation

UVA might be useful for increasing nicotine content, but the therapeutic window is narrow.[24]

Harvest

Post-Harvest Processing

Herb scissors.

Yield

Seed production is adversely affected by damage (including leaf picking and suckering). However, that effect can be mitigated by hormonal application.[26]

Nicotine does not degrade in sunlight.[27]

[23] [28] [29] [30] [31] [32] [33] [34] [35] [11] [36] [37] [38] [39] [40] [2] [41] [16] [42] [43] [44] [45] [46] [47] [48] [49] [50] [51] [20] [52] [53] [54]

[2]

Soilless

Rustica can be grown in Hoagland hydroponic solution.[22]

Soil

Fertilization

In a study of various ratios of urea to ammonium sulfate with a constant nitrogen application rate (200 kg/ha), between zero and twenty percent ammonium sulfate had the highest cured leaf yield while maintaining nicotine concentration. This may have been an effect of correcting the marginally sulfur-deficient soil (15.25 ppm).[25]

Nitrogen source does not significantly affect nicotine concentration, only growth.[26][25][23]

To reduce the application rate of expensive farmyard manure, Arya et. al found that the manure dose could be lowered to 2.5 t/ha while maintaining a high-profit margin if Azotobacter and phosphorus solubilizing bacteria are added. However, the full dose of all fertilizers studied (N, P, K, and manure) still produced the highest yield, cost/benefit ratio, and first-grade leaf yield, therefore, this reduced application rate should only be used in cases of manure scarcity.[23]

Rustica growth suffers at a salt concentration of 25mM, therefore rustica is a salt-sensitive glycophyte species. A salt concentration of 75 mM or higher was sufficient to cause death. Silicon in the form of 1mM sodium silicate application was able to ameliorate some of the negative effects of high salinity and even improved growth beyond the control group without salt.[22]

Temperature

Lighting

Pests

Ethylene is involved in reducing the damage-induced increase in nicotine production.[55]

Leaf damage is exacerbated by competition with other plants in the same soil or hydroponic media.[26]

Morphology

Stem

Leaves

Inflorescence

Seeds

Phytochemistry

Infraspecific Variation

Biosynthesis

Root tips are the source of nicotine. Root-bound potted plants will not produce excess nicotine in response to simulated herbivory.[56]

Distribution

Timecourse

Rustica emits more scented compounds at night primarily as benzenoids.[57]

Improvement

Identification

Inheritance

Methods

Nicotiana does not exhibit parthenogenesis. However, nicotiana species can stimulate the seed production of others, but the resultant offspring are not hybrids. Rather they are true to the mother type and sterile.[58]

History & Society

Work Log

15 Jul 2023

14 Jul 2023

11 Jul 2023

Bibliography

1. Cooper, Valerie C. and Walkey, D. G. A., 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.
2. Matusiewicz, E., 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. url: 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...
3. Pearce, Robert C. and Li, Yongmei and Bush, Lowell P., 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, July 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.
4. Saha, S. and Modak, S. B. and Basu, P. K., 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. url: 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...
5. Steinberg, Robert A., Production and Germination of Nicotiana Seed Grown Under Various Greenhouse Conditions, Tobacco Science, vol. 216, pp. 4--46, 1960.
6. Zaghdoud, Chokri and {Maâroufi-Dguimi}, Houda and Ouni, Youssef and Guerfel, Mokhtar and Gouia, Houda and Negaz, Kamel-Eddine and Ferchichi, Ali and Debouba, Mohamed, 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
7. Boughey, H. J., Competitive Ability and Environmental Sensitivity in Nicotiana Rustica., 1978. url: 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.
8. Breese, E. L., Selection for Differing Degrees of Out-Breeding in Nicotiana Rustica, Annals of Botany, vol. 23, no. 2, pp. 331--344, April 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.
9. Chowdhury, M. K. U., An Improved Method for Dihaploid Production in Nicotiana Rustica through Anther Culture, Theoretical and Applied Genetics, vol. 69, no. 2, pp. 199--204, December 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.
10. Dudek, M., 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. url: 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...
11. Essa, A. K., Variations in Alkaloidal Constituents of Plant Tissue Cultures. [Nicotiana Tabacum, Nicotiana Rustica, Datura Stramonium and Hyoscyamus Niger], January 1987. url: 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.
12. Gill, Ravinder and Rashid, A. and Maheshwari, S. C., 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.
13. Hamill, J. D. and Pental, D. and Cocking, E. C., 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, December 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.
14. Handique, A. K. and Sharma, C. M., Colchicine Induced Hypertrophy in Catharanthus Roseus (L.) G. Don and Nicotiana Rustica L., Crop Research (Hisar), vol. 3, no. 2, pp. 287--290, 1990. url: 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....
15. Jinks, J. L. and Pooni, H. S., Determination of the Environmental Sensitivity of Selection Lines of Nicotiana Rustica by the Selection Environment, Heredity, vol. 49, no. 3, pp. 291--294, December 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.
16. Matusiewicz, E., 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. url: 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....
17. Pooni, H. S. and Foster, R. and Zhao, B., 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, March 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.
18. Saxena, S. S. and Tyagi, I. D., 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. url: 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.
19. Tabata, Mamoru and Hiraoka, Noboru, 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.
20. Waratadar, Asif and Nirmalnath, P. Jones and Matiwade, P. S. and Kr, Abhinandana and Ks, Raghavendra, 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. url: 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.
21. Werner, C. P. and Dunkin, I. M. and Cornish, M. A. and Jones, G. H., Gene Transfer in Nicotiana Rustica by Means of Irradiated Pollen II Cytogenetical Consequences, Heredity, vol. 52, no. 1, pp. 113--119, February 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.
22. Hajiboland, R. and Cheraghvareh, L., 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, September 2014. url: 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.
23. Arya, R. L. and Kumar, P. Harishu and Roy, S. and Chandrasekhararao, C. and Amarnath, S. and Chanda, S., Effect of Integrated Nutrient Management on Yield, Quality, and Economics of Motihari I Tobacco (Nicotiana Rustica) in Terai Region of North Bengal, June 2011. url: 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.
24. Kartusch, Robert and Mittendorfer, Bettina, Ultraviolet Radiation Increases Nicotine Production in Nicotiana Callus Cultures, Journal of Plant Physiology, vol. 136, no. 1, pp. 110--114, April 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.
25. Parmar, S C and Rathod, S V and Gondaliya, S M and Chaudhary, N N, 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. url: 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.
26. Baldwin, Ian T., Inducible Nicotine Production in Native Nicotiana as an Example of Adaptive Phenotypic Plasticity, Journal of Chemical Ecology, vol. 25, no. 1, pp. 3--30, January 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.
27. Ohta, Shinsuke and Yatazawa, Michihiko, Effect of Light on Nicotine Production in Tobacco Tissue Culture, Agricultural and Biological Chemistry, vol. 42, no. 4, pp. 873--877, April 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.
28. Bocciarelli, Dorian, Medium Optimization for the Rhizosecretion of Three Monoclonal Antibodies from Nicotiana Tabacum Hydroponic Culture, pp. 29, September 2014. url: 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.
29. Bouthour, D. and {Hajjaji-Nasraoui}, A. and Saafi, L. and Gouia, H. and {Chaffei-Haouari}, C., 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)
30. Bridgen, Mark P., Studies of in Vitro Flowering and de Novo Flowers of Nicotiana Tabacum, 1984. url: 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.
31. Chandnani, J. J. and Thomas, A. I., Preliminary Studies on Topping and Suckering in Hookah Tobacco (Nicotiana Rustica)., Indian Journal of Agronomy, vol. 4, pp. 144--7, 1960. url: 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...
32. Chaubey, C. N. and Mishra, S. K. and Mishra, A. P., Study of Variability and Path Analysis for Leaf Yield Components in Hookah Tobacco., Tobacco Research, vol. 16, no. 1, pp. 47--52, 1990. url: 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.
33. Dimitrijevic, R., 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. url: 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.
34. Douglas, G. C. and Keller, W. A. and Setterfield, G., 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, February 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.
35. Drake, Katherine E. and Moore, J. Michael and Bertrand, Paul and Fortnum, Bruce and Peterson, Paul and Lewis, Ramsey S., 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.
36. Ghader, Habibi and Zienab, Sadeghipour and Roghieh, Hajiboland, Effect of Salicylic Acid on Tobacco (Nicotiana Rustica) Plant Under Drought Conditions, vol. 7, no. 25, pp. 17--28, January 2015. url: https://www.sid.ir/paper/159996/fa. Abstract
    {$<$}P dir=ltr align=left{$>$}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\textsubscript{2} 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.{$<$}?xml:namespace prefix = o ns = urn:schemas-microsoft-com:office:office /{$><$}o:p{$><$}/o:p{$>$}{$<$}/P{$>$}
37. Hunt, Warren F. and Loomis, Robert S., 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.
38. Katba, P. J. and Pandor, B. L. and Gediya, L. N., Genetic Association among Different Characters in Tobacco (Nicotiana Rustica L.), Journal of Pharmacognosy and Phytochemistry, vol. 8, no. 1, pp. 146--148, 2019. url: 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.
39. Katba, P. J. and Hadiya, R. G. and Kapadia, V. N. and Patel, D. C. and Patel, A. D., 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. url: 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.
40. Laszlo, Csaba and Kaminski, Kacper and Guan, Haifeng and Fatarova, Maria and Wei, Jianbing and Bergounioux, Alexandre and Schlage, Walter K. and {Schorderet-Weber}, Sandra and Guy, Philippe A. and Ivanov, Nikolai V. and Lamottke, Kai and Hoeng, Julia, Fractionation and Extraction Optimization of Potentially Valuable Compounds and Their Profiling in Six Varieties of Two Nicotiana Species, Molecules, vol. 27, no. 22, pp. 8105, January 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.
41. Matusiewicz, E., Obtaining nicotine from Nicotiana rustica., Tyton, Warszawa, vol. 6, no. 9-10, pp. 6--7, 1950. url: 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.
42. Mench, M. and Tancogne, J. and Gomez, A. and Juste, C., 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, July 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.
43. Ninama, {\relax SD} and Gediya, {\relax KM} and Rathwa, {\relax MK} and Vaghela, {\relax GM}, 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. url: 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.
44. Panchal, Jalpa and Gedia, K. M., 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. url: 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...
45. Parajuli, Suman and Patel, D A and Makwana, M G and Parmar, D J, Genetic Diversity Studies on Cured Leaf Yield and Its Components in Rustica Tobacco [Nicotiana Rustica (L.)], pp. 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.
46. Pateli, H N, Effect of Biparental Mating and Extent of Genetic Variability in Tobacco (Nicotiana Rustica L.), Green Farming, vol. 1, no. 2, pp. 158--160, March 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.
47. Popova, Venelina and Ivanova, Tanya and Stoyanova, Albena and Nikolova, Violeta and Hristeva, Tsveta and Zheljazkov, Valtcho D., GC-MS Composition and Olfactory Profile of Concretes from the Flowers of Four Nicotiana Species, Molecules, vol. 25, no. 11, pp. 2617, January 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.
48. Prajapati, R B and Gediya, K M and Patel, D M, Feasibility of Vegetable Intercropping in Rustica Tobacco (Nicotiana Rustica L.) Under Middle Gujarat Conditions, Advances in Life Sciences, pp. 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.
49. Prather, Charles W. and Sisler, Edward C., 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.
50. Röper, W. and Schulz, M. and Chaouiche, E. and Meloh, K. A., Nicotine Production by Tissue Cultures of Tobacco as Influenced by Various Culture Parameters, Journal of Plant Physiology, vol. 118, no. 5, pp. 463--470, April 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.
51. Sabour, M. and Simmonds, J. and Setterfield, G., 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.
52. Woras, G. and Hashmi, E. A. and Karim, F. and Shahid, M. and Shah, S. A., Comparative Evaluation of Some Tobacco Varieties (Nicotiana rusticaL.), Pak Tobacco (Pakistan), 2004. url: 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.
53. Zhang, Bo and Wang, Xiao-qin and Li, Xin and Ni, Yong-qing and Li, Hong-yu, 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.
54. Zhao, Bo and Agblevor, Foster A. and K. C., Ritesh and Jelesko, John G., 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, April 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.
55. Kahl, Johannes and Siemens, David H. and Aerts, Rob J. and Gäbler, Ralph and Kühnemann, Frank and Preston, Catherine A. and Baldwin, Ian T., 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, January 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.
56. Baldwin, Ian T., Damage-Induced Alkaloids in Tobacco: Pot-Bound Plants Are Not Inducible, Journal of Chemical Ecology, vol. 14, no. 4, pp. 1113--1120, April 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.
57. Raguso, Robert A and Levin, Rachel A and Foose, Susan E and Holmberg, Meredith W and McDade, Lucinda A, Fragrance Chemistry, Nocturnal Rhythms and Pollination “Syndromes” in Nicotiana, Phytochemistry, vol. 63, no. 3, pp. 265--284, June 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.
58. Wellington, Richard, 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.