Analyses and evaluation of the main chemical components in different tobacco (Nicotiana tabacum L.) genotypes

2.1. Analyses of tobacco leaves

2.2. Analyses of tobacco seeds

2.3. Statistical analysis

The nicotine, reducing sugar, ion contents and seed crude oil determinations were repeated three times, and their analyses were performed within each replicate sample three times with standard deviation (SD). Significant differences among the genotypes were determined by the one-way ANOVA with means separation by the Least Significant Difference (LSD) test at the 0.01 level. dendrogram was used to show the hierarchy of clusters and to determine the genetic variability among tobacco cultivars and genotypes based on chemical components and FAC by the XLSTAT program (Yaldiz and Camlica, 2019Yaldiz G, Çamlica M. 2019. Variation in the fruit phytochemical and mineral composition, and phenolic content and antioxidant activity of the fruit extracts of different fennel (Foeniculum vulgare L.) genotypes. Ind. Crops Prod. 142, 111852. https://doi.org/10.1016/j.indcrop.2019.111852 ).

3.1. Nicotine content (NC)

NC is the most important factor in the tobacco industry especially in cigarette blends, despite the fact that it has some negative effects on the human body as healthy and protective. The quality parameter of the cigarette blend is determined by the NC. There were statistically significant differences among the tobacco genotypes and cultivar in terms of the NC (Table 3). The NC of tobacco genotypes and cultivar varied between 0.10-0.87%. The highest NC was found for the ESR-4 genotype when compared to the others, followed by ESR-5 (0.78%) and BSR-5 (0.59%). Cultivar ‘Akhisar 97’ was found to be the highest of the 20 genotypes at 0.41%. It was found that MR-4 (0.10%) and MR-3 (0.16%) genotypes contained lower nicotine ratios than the others. While the highest values were obtained for ESR-4 and ESR-5, the lowest values were obtained from MR-2, 3, 4, and 5 genotypes. Compared to the average NC of the tobacco-growing regions, AR, MR, ESR, and BSR were found at 0.33, 0.22, 0.46, and 0.37%, respectively. The results showed that the tobacco genotypes of ESR were higher than the others. This situation can be explained by the ecological conditions of Bolu, which are similar to a continental climate. It was noted that the characterization of quality and aroma of the tobacco depended on the soil, climate conditions and low amount of nitrogen (Bilgin et al., 1993Bilgin AE, Müftüoğlu Y, Ustralı A. 1993. Calibration of Commercial Tobacco Requirements and Phosphorus-Potash Analysis Methods of Oriental Tobacco Under Field Conditions in Aegean Region. T. C. Ministry of Agriculture and Rural Affairs General Directorate of Rural Services Menemen Research Institute Publications. Publication No: 1995, İzmir. 1993.). It was also reported that Aegean tobacco had very low nicotine contents (Delibacak et al., 2014Delibacak S, Ongun AR, Ekren S. 2014. Influence of soil properties on yield and quality of tobacco plant in Akhisar region of Turkey. Eurasian J. Soil Sci. 3, 286-292.). Abdallah (1986)Abdallah F. 1986. Is Tobacco Quality Measurable? Trans. K. Ketenci. Monopoly Institute Istanbul. reported that high levels of nicotine gave the hard and burner features, and low levels of nicotine led to poor taste and physiological dissatisfaction. These tobaccos can be used because of their low nicotine level and their rich flavor (Otan and Apti, 1989Otan H, Apti R. 1989. Tobacco. T.C. T.O.K.İ.B. Ege Agricultural Research Institute Publications, No: 83. Menemen-İzmir.). Although in low doses nicotine has a stimulating effect, increasing activity, alertness and memory, it also increases the heart rate and blood pressure and causes anorexia (Bastida and Beltran, 2011Bastida G, Beltran B. 2011. Ulcerative colitis in smokers, non-smokers and ex- smokers. World J. Gastroenterol. 17, 2740-2747. https://doi.org/10.3748/wjg.v17.i22.2740 ). These tobaccos can be used in the tobacco industry for protect human health because of their low NC.

Ekren and Sekin (2008)Ekren S, Sekin S. 2008. Investigation of chemical and expertise characteristics of Akhisar region tobaccos and relations with the leaf yield. Ege J. Agric. Res. 45 (3), 165-173. noted that NC was found from 0.12 to 1.06% in tobacco from the Akhisar region. 1.8-2.1% (Kurt and Ayan, 2014Kurt D, Ayan AK. 2014. Effect of the different organic fertilizer sources and doses on yield in organic tobacco (Nicotiana tabacum L.) production. J. Agric. Faculty Gaziosmanpasa Univer. 31 (2), 7-14.), 0.73-0.95% (Yagac, 2015Yagac C. 2015. Determination of yield and quality characters of tobacco cultivars of Aegean Region in Denizli conditions. MsC Namık Kemal University Graduate School of Natural and Applied Sciences Department of Field Crops.) were also found.

The results were similar to those of other researchers, except for Kurt and Ayan (2014)Kurt D, Ayan AK. 2014. Effect of the different organic fertilizer sources and doses on yield in organic tobacco (Nicotiana tabacum L.) production. J. Agric. Faculty Gaziosmanpasa Univer. 31 (2), 7-14.. The differences can be explained by the fact that the tobacco genotypes and cultivar showed differences depending on their adaptation, genetic properties and survival in different places and the soil contents as organic matter can affect the NC of tobacco (Griesser et al., 2015Griesser M, Weingart G, Schoedl-Hummel K, Neumann N, Becker M, Varmuza K, Liebner F, Schuhmacher R, Forneck A. 2015. Severe drought stress is affecting selected primary metabolites, polyphenols, and volatile metabolites in grapevine leaves (Vitis vinifera cv. Pinot noir). Plant. Physiol. Biochem. 88, 17-26. https://doi.org/10.1016/j.plaphy.2015.01.004 ).

3.2. Reducing sugar content (RSC)

Reducing sugar contents can provide mildness when smoking tobacco and was found to positively affect its quality (Abdallah 1986Abdallah F. 1986. Is Tobacco Quality Measurable? Trans. K. Ketenci. Monopoly Institute Istanbul.). Differences in RSC were found to be statistically significant among the tobacco genotypes and cultivar. As seen in Table 3, the RSC of tobacco genotypes and cultivar ranged from 9.70 to 21.30%. The RSC of ESR-9 and ESR-11 were higher than the other genotypes and cultivar. The ESR-2, ESR-5 and ESR-10 genotypes were found to be lower than the others at 9.70%. When compared among the regions, the highest RSC was found in MR at 16.70% followed by AR (15.43%), ESR (14.48%) and BSR (14.04%). The cultivar was found to 16.90% above the average of the regions.

The desirable amount of RSC is between 8-13% (Camas et al., 2007Camas N, Karaali H, Ozcan H. 2007. Effects of different fertilizer doses on yield, quality and technological properties of pressed tobacco genotype in Erbaa-Taşova Conditions. 2007 TTL Foreign Trade. Research Report.). The closest values to the desired ones were seen in 8 genotypes between 9.70-13.0%. The values were found in BSR genotypes (BSR-2, 3, 4, 5), followed by ESR genotypes (ESR-2, ESR-5 and ESR-10) and MR-1 in MR genotypes. Based on the RSC values (Table 3), these genotypes are different from the others. Therefore, they are suitable for the production of the RSC used in the tobacco industry for a quality product.

It is noted that the RSC of tobacco varied between 11.30-21.83% (Yagac, 2015Yagac C. 2015. Determination of yield and quality characters of tobacco cultivars of Aegean Region in Denizli conditions. MsC Namık Kemal University Graduate School of Natural and Applied Sciences Department of Field Crops.) and 7.81-33.71 (Ekren and Sekin, 2008Ekren S, Sekin S. 2008. Investigation of chemical and expertise characteristics of Akhisar region tobaccos and relations with the leaf yield. Ege J. Agric. Res. 45 (3), 165-173.). Our results have been compared to those of other researchers and it was determined that they showed similar values as reported by researchers.

3.3. Ion content (IC)

Ions are non-combustible substances and are only present in the ash. It is known that, due to the increase in potassium from these substances, the ability of the tobacco to burn improves, while the chloride salt affects it negatively (Er and Yıldız 2014Er C, Yıldız M. 2014. Pleasure plant. Ankara University Publications, No: 419.).

The present study was conducted for the evaluation of IM such as Potassium (K+), Magnesium (Mg²⁺), Calcium (Ca²⁺), Chloride (CI^-), Phosphorus (PO₄ ^3-) and Sulfate (SO₄ ^2-), and in the seeds of tobacco genotypes and cultivar. Significant differences were found among tobacco genotypes and cultivar in terms of IC (Table 4).

In this study, the concentration of K⁺ varied from 8.1 to 52.6 mg/g plant. The K⁺ concentrations in the leaves were different among all the different genotypes. The highest value was found for BSR-5 and the lowest value was found for BSR-7. Krishnamurthy and Ramakrishnayya (1993) reported that the tobacco plant had a higher K requirement compared to other cultivated plants. They also noted that color, texture, thickness, elasticity, and burning capacity were affected by dry leaves and the high K⁺ content in the dried leaves was generally considered to be the highest quality criterion. The obtained K⁺ values were higher than those reported by Irget et al., (1999)Irget ME, Oktay M, Hakerlerler H, Atil H, Cakici H. 1999. A research on the nutrition and soil-plant relationships of Virginia (flue-cured) tobacco grown in Düzce. J. AARI 9 (2), 125-142. (0.87-2.24%). Our tobaccos were found to be rich in terms of K⁺. Therefore, especially BSR-5 and ESR-5 genotypes were found to be more suitable for the burning capacity of tobacco.

The Mg²⁺ concentrations of the tobacco genotypes and cultivar varied between 6.04 and 0.61 mg/g plant (Table 4). The highest value was determined for BSR-3, and the lowest value was determined for the AR-4 genotype. Ca²⁺ content varied from 0.27 to 37.4 mg/g. While the highest values were obtained for ESR-10 (37.4 mg/g) and BSR-1 (34.7 mg/g), the lowest values were obtained for the ESR-4 (0.27 mg/g) and BSR-10 (10.7 mg/g) genotypes. Cl^- was present in the range of 1.28-32.3 mg/g. The highest concentration was present in the BSR-6 genotype, followed by the BSR-3 (29.2 mg/g) genotype. In the present study, the concentration range of PO₄ ^3- was 4.29-9.44 mg/g, as shown in Table 4. The highest level of that form of PO₄ ^3- was found in the ESR-7 genotype (9.44 mg/g), followed by the BSR-6 genotype (8.38 mg/g). SO₄ ^2- concentrations ranged from 2.61 to 7.40 mg/g. Its maximum content (7.40mg/g) was present in ESR-5, and its minimum content (2.61 mg/g) was present in BSR-9. The highest amounts of K⁺, Mg²⁺ and Ca²⁺ were found in the MR genotypes, the highest PO₄ ^3- and SO₄ ^2- in the ESR genotypes and the highest CI^- was seen in the BSR tobaccos. The change in the same ion contents in the tobacco genotypes among the regions can be explained by genetic differences and tobacco producers may have used the same tobacco genotype seeds in the same region.

Ca²⁺, Mg²⁺ and CI^- contents were found between 2.08-6.32%, 0.20-0.98%, and 0.1-0.50%, respectively (Irget et al., 1999Irget ME, Oktay M, Hakerlerler H, Atil H, Cakici H. 1999. A research on the nutrition and soil-plant relationships of Virginia (flue-cured) tobacco grown in Düzce. J. AARI 9 (2), 125-142.). The obtained data was found somewhat similar to other researchers.

3.4. Seed crude oil (SCO)

Significant differences were found among the tobacco genotypes and cultivar in terms of SCO, as shown in Table 3. The SCO values ranged from 24.33-47.0%. The analysis showed that SCO was the highest in BSR-5 (47.0%), followed by BSR-6 (46.33%) genotypes and AR-1 cultivar (46.33%). The lowest SCO values were found for the ESR-2 (24.33%), MR-3 (25.33%) and AR-4 (25.33%) genotypes. When the SCO of the tobacco genotypes and cultivar were compared among the regions, SCO was found at 32.44% in AR, 30.20% in MR, 38.06% in ESR, and 39.20% in BSR. In general, the average SCO of the tobacco genotypes and cultivar was over 30%, with the highest average SCO determined for the BSR genotypes. So, BSR-5, BSR-6 and the AR-1 cultivar had highest crude oil contents and they can be used in biodiesel production as an alternative fuel source (Fornasier et al., 2018Fornasier F, Gomez JFC, de Carvalho FS, Schneider RdCdS, Costa ABd, Moraes JAR, Bravo dCAG. 2018. Biodiesel production from energy tobacco. Orbital: Electronic J. Chem. 10 (2), 123-132.).

Tobacco seed oil was reported as 24.56-41.93% (Mohammad and Tahir 2014Mohammad MT, Tahir NA. 2014. Evaluation of chemical compositions of tobacco (Nicotiana tabacum L) genotypes. Seeds Annual Res. Review Biol. 4 (9), 1480-1489. https://doi.org/10.9734/ARRB/2014/8211 ), 29.82% (Abbas Ali et al., 2008Abbas Ali M, Abu Sayeed M, Roy RK. 2008. Comparative study of on characteristics of Seeds oils and Nutritional composition of seeds from different varieties of tobacco cultivated in Bangladesh. Asian J. Biochem. 3 (4), 203-212. https://doi.org/10.3923/ajb.2008.203.212 ). The obtained data were found similarly those reported by other researchers.

3.5. Fatty acid compositions (FAC)

Table 5 shows the FAC of the tobacco genotypes and cultivar. Linoleic (18:2), oleic (18:1), palmitic (C16:0) and butyric acids (C4:0) were the four most abundant acids among the 25 FAC (13.92-75.04, 0.46-17.80, 5.55-19.11 and 0.33-64.98%, respectively).

Linoleic acid was found to be the main component and predominated the oils of tobacco (13.92-75.04%), except for the MR-5 and ESR-6 genotypes. The highest linoleic acid was found in ESR-3 (75.04%), followed by the AR-1 cultivar (67.84%) and AR-4 (67.44%) genotype. The lowest linoleic acid was determined for BSR-8 (13.92%) and ESR-9 (24.54%). The high content of linoleic acid in tobacco seed oil is very important for the production of oleo-chemicals (Abbas Ali et al., 2008Abbas Ali M, Abu Sayeed M, Roy RK. 2008. Comparative study of on characteristics of Seeds oils and Nutritional composition of seeds from different varieties of tobacco cultivated in Bangladesh. Asian J. Biochem. 3 (4), 203-212. https://doi.org/10.3923/ajb.2008.203.212 ). It can be used as a surfactant, dispersant, bio lubricant, in cosmetics and a variety of synthetics in the formulation of protective coating and in preparations of other long chain compounds (Awola et al., 2010Awola VG, Ogunniyi DS, Odetoye TE. 2010. Refining, modification and characterization of tobacco seed oil for improved potentials for industrial use. Nigerian J. Pure Appl. Sci. 2168-2174.). The content of linoleic acid was higher than that reported (4.2-9.23%) by Mohammad and Tahir (2014)Mohammad MT, Tahir NA. 2014. Evaluation of chemical compositions of tobacco (Nicotiana tabacum L) genotypes. Seeds Annual Res. Review Biol. 4 (9), 1480-1489. https://doi.org/10.9734/ARRB/2014/8211 and found similar to the 6.45-77.48% reported by Poltronieri (2016)Poltronieri P. 2016. Tobacco seed oil for biofuels. Pp. 161-185. In: Poltronieri P and D’Urso O.F. Biotransformation of Agricultural Waste and By-Products. The Food, Feed, Fibre, Fuel (4F) Economy. Elsevier, Oxford, UK. 2016. ISBN: 9780128036228. https://doi.org/10.1016/B978-0-12-803622-8.00006-9 . Therefore, especially the ESR-3 and AR-1 genotypes are promising for linoleic acid to be obtained at high levels and these genotypes can be used in oleo-chemical production and other areas such as cosmetic and surfactant. It can also be used in making quick-drying oils such as oil paints and varnishes (Chiririwa et al., 2014Chiririwa H, Hapanyengwi A, Muzenda E. 2014. Tobacco seed oil as an economically viable alternative for the tobacco ındustry. International Conference on Chemical Engineering and Advanced Computational Technologies, November, 24-25, 36-39. https://doi.org/10.15242/IIE.E1114017 ). In addition to this, Kirkova et al., (2016)Kirkova S, Srbinoska M, Ivanova S, Georgieva A. 2016. Determination of fatty acid composition of seed of oriental tobacco. Tobacco 66 (1-6), 53-58. https://doi.org/10.13140/RG.2.2.18885.14560 noted that the linoleic acid content must be lower to obtain better oil as quality. Therefore, the BSR-8 and ESR-9 genotypes can be evaluated for the best oil quality.

High concentrations of oleic (0.46-17.80%) and palmitic acids were detected (5.55-19.12%; except for ESR-1 genotype) in all the tobacco studied oils. The highest oleic acid was found in the BSR-3 genotype with 17.80%, followed by ESR-9 (17.35%) and AR-2 (16.35%). The lowest oleic acid was found in ESR-8 (0.46%). The highest palmitic acid was determined in MR-5 (19.12%) and the lowest palmitic acid was found in the BSR-7 genotype.

The effects of oleic acid are mediated by preventing the reduction in palmitic acid-mediated activated protein kinase (AMPK) activity, resembling the action of metformin (Palomer et al., 2018Palomer X, Pizarro-Delgado J, Barroso E, Vázquez-Carrera M. 2018. Palmitic and oleic acid: The yin and yang of fatty acids in type 2 diabetes mellitus. Trends Endocrinol. Metabol. 29 (3), 178-190. https://doi.org/10.1016/j.tem.2017.11.009 ). Palmitic and oleic acids were determined at 21.33-25.667% and 17.00-26.667% by Mohammad and Tahir (2014)Mohammad MT, Tahir NA. 2014. Evaluation of chemical compositions of tobacco (Nicotiana tabacum L) genotypes. Seeds Annual Res. Review Biol. 4 (9), 1480-1489. https://doi.org/10.9734/ARRB/2014/8211 . The main fatty acids (FA) in tobacco were found to be linoleic acid (60-80%), oleic acid (10-20%) and palmitic acid (8-20%) (Giannelos et al., 2002Giannelos PN, Zannikos F, Stourna S, Lois E, Anastopoulos G. 2002. Tobacco seed oil as an alternative diesel fuel: physical and chemical properties. Ind. Crops Prod. 16, 1-9. https://doi.org/10.1016/S0926-6690(02)00002-X ; Abbas Ali et al., 2008Abbas Ali M, Abu Sayeed M, Roy RK. 2008. Comparative study of on characteristics of Seeds oils and Nutritional composition of seeds from different varieties of tobacco cultivated in Bangladesh. Asian J. Biochem. 3 (4), 203-212. https://doi.org/10.3923/ajb.2008.203.212 ; Stanisavljević et al., 2009Stanisavljević IT, Veličković DT, Todorović ZB, Lazić ML, Veljković VB. 2009. Comparison of techniques for extraction of tobacco seed oil. Europ. J. Lipid Sci. Technol. 111 (5), 513-518. https://doi.org/10.1002/ejlt.200800232 ; Bucciarelli et al., 2013Bucciarelli S, Civitella D, Fecondo G, Ghiann G, Sorrentino C, Del Piano L, Raimo F, Stanisci V. 2013. Influence of plant density and leaf priming on seed and seed oil yield tobacco. Proc. CORESTA Meeting, Agronomy/Phytopathology, Brufa di Torgiano (PG), Italy.; del Piano et al., 2014Piano L del, Abet M, Raimo F, Modestia F, Sicignano M, Enotrio T. 2014b. Caraterizzazione morfologica e qualitativa di accessioni di Nicotiana tabacum L. per la produzione di olio da seme. Proc. X National Congress on Biodiversity, Rome, Italy.).

The palmitic and oleic acid results were found to be different from those obtained by Mohammad and Tahir (2014)Mohammad MT, Tahir NA. 2014. Evaluation of chemical compositions of tobacco (Nicotiana tabacum L) genotypes. Seeds Annual Res. Review Biol. 4 (9), 1480-1489. https://doi.org/10.9734/ARRB/2014/8211 ; but they were close to those reported by other researchers. It has been reported that climatic conditions affect the properties of plants such as growth, yield and biochemical contents (Zandalinas et al., 2018Zandalinas SI, Mittler R, Balfagón D, Arbona V, Gómez-Cadenas A. 2018. Plant adaptations to the combination of drought and high temperatures. Physiol. Plant. 162, 2-12. https://doi.org/10.1111/ppl.12540 ).

Butyric acid can be used in different industries, and there has recently been a great interest in using it as a precursor to biofuels (Dwidar et al., 2012Dwidar M, Park JY, Mitchell RJ, Sang B. 2012. The future of butyric acid in ındustry. Scient. World J. 1-9. https://doi.org/10.1100/2012/471417 ). Moreover, it has many uses in the pharmaceutical and chemical industries. It was well known for its anticancer effects and it induces morphological and biochemical changes in various cells leading to the concomitant suppression of neoplastic properties (Cao et al., 2011Cao Y, Li HQ, Zhang J. 2011. Homogeneous synthesis and characterization of cellulose acetate butyrate (CAB) in 1-Allyl-3-methylimidazolium chloride (AmimCl) ionic liquid. Indust. Engineer. Chem. Res. 50 (13), 7808-7814. https://doi.org/10.1021/ie2004362 ). Butyric acid was found between 0.33-64.98% in all the tobaccos. The highest butyric acid was found for ESR-6 (64.98%), followed by the MR-5 (58.61%) and BSR-7 (35.54%) genotypes. The lowest butyric acid was found in ESR-3 with 0.33%. ESR-6, MR-5 and BSR-7 genotypes can be cultivated to obtain high butyric acid contents for use in biofuel.

A correlation was seen between butyric acid and linoleic acid. When linoleic acid was not detected, such as in ESR-6 and MR-5, butyric acid was found at the highest levels in these genotypes. Elaidic and cis-11, 14 heneicosanoic acids were the minor fatty acids in these tobaccos, constituting 0.04-7.22 % and 0.04-1.97%, respectively. Linoleic acid and butyric acid were found at the highest FA rates among the others at 54.0% and 18.57%, respectively.

The FA of tobacco seed crude oil can be divided into two main components as saturated fatty acids (SFA) and unsaturated fatty acids (USFA). The total SFA and USFA in tobacco seed oil were found to be 33.65 and 64.26%, respectively. While the 11 FA compositions were found as SFA, 14 FA compositions were determined as USFA. SFA compositions varied from 11.51 to 88.84% in ESR-3 and ESR-6 from the ESR genotypes. The USFA compositions were found between 11.16-87.98% in ESR-3 and ESR-6 in the ESR genotypes. Among the USFA, linoleic acid (54.0%) and oleic acid (9.52%) were the most abundant while palmitic and butyric acids (18.71%) were most abundant among the SFA (Table 5).

The total FA compositions ranged from 77.94-100% in the tobacco genotypes and cultivar. The highest total FA compositions were found in the ESR-1 and BSR-8 genotypes (100%), followed by MR-5, ESR-6 and BSR-10 with 99.99%. The lowest total FA compositions were seen in BSR-6 (77.94%), followed by the BSR-3 (94.42%) and MR-2 (94.69%) genotypes.

The amounts of unsaturated and saturated fatty acids reported in the present work are close to those reported as 73.95-26.1% (Zalatanov et al., 2000Zalatanov M, Menkov N. 2000. Phospholipid and fatty acid composition of tobacco seeds. Rostlinna Vyroba 46, 439-441.) and 85.2 and 14.8% (Giannelos et al., 2002Giannelos PN, Zannikos F, Stourna S, Lois E, Anastopoulos G. 2002. Tobacco seed oil as an alternative diesel fuel: physical and chemical properties. Ind. Crops Prod. 16, 1-9. https://doi.org/10.1016/S0926-6690(02)00002-X ), respectively.

3.6. Genetic diversity of tobacco cultivar and genotypes

Dendrogram analyses were carried out among the chemical components as NC, RSC, SCO and 25 FACs in two different figures in 29 tobacco genotypes and 1 cultivar. The dendrogram analyses were divided into two main groups in terms of NC, RSC, SCO and FAC as A and B (Figures 1a and b). All figures consisted of 2 sub-groups as A1 and B1. While group A was formed of 16 genotypes and one cultivar, group B contained 13 genotypes. Sub-group A1 comprised 11. including cultivar (AR-1), 4 ESR, 4 BSR, 1 MR and 1 AR genotypes. Sub-group A2 included 6 genotypes (5 ESR, 5 BSR, 2 AR and 1 MR genotypes). 3 genotypes fell into sub-group B1 (ESR 4, ESR 5 and BSR 5) and sub-group B2 had 10 genotypes as 4 BSR, 3 ESR, 2 AR and 1 AR genotypes (Figure 1a). When figure 1b was examined, it was seen that the FACs of the tobacco cultivar and genotypes were divided into four sub-groups as A1, A2, B1 and B2. Sub-group A1 and B1 had only one FAC as lignoceric acid and arachidic acid, respectively. While group A2 had 13 FACs including one of the major acids, butyric and linoleic acid; B2 had FACs. Figures 1a and 1b were evaluated together and the major FACS were determined in group A and B1 subgroup and minor FACs were observed in B2 in Figure 1b was formed based on linoleic acid. Most of the saturated acids were found in group A and unsaturated acids were detected in group B in Figure 1b.