Evaluation of genetic diversity of some tea genotypes attributed to Darjeeling in Iran using ISSR markers

Document Type : scientific research article

Authors

1 Assistant Prof., Tea Research Center, Horticultural Sciences Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Lahijan, Iran.

2 Corresponding Author, Assistant Prof., Tea Research Center, Horticultural Sciences Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Lahijan, Iran.

Abstract

Background and objectives: Tea (Camellia sinensis (L.) O. Kuntze) is the most consumed beverage in the world after water and an important commercial plant with economic value in northern Iran. Over the years, various tea plants have been cultivated in Iran's tea gardens by seed method, which has led to a high diversity in them. Since most gardeners have propagated the plant sexually, tea gardens vary in quality. Therefore, determining the genetic diversity and relatedness of plants has an important role to support breeding and breeding programs.

Materials and methods: In this study, 28 tea plants attributed to Darjeeling region along with eight Iranian samples and eight samples imported from Sri Lanka were investigated for genetic diversity using ISSR markers. After sampling from young and fully developed leaves, their genomic DNA was extracted and 16 ISSR markers were used to investigate the genetic relationships of 44 tea samples. The obtained data were analyzed by the simple similarity coefficient for the ISSR markers and the cluster was designed based on the UPGMA algorithm. Population structure analysis was also done by POPGENE program.

Results: The use of 16 ISSR primers produced 158 bands, of which 116 bands showed polymorphism, and based on this, the percentage of polymorphism was calculated as 73.42%. The PIC test showed a range of 0.45 to 0.50 and this indicator was 0.49 for all markers. The results of Cophentic test showed that SM similarity coefficient and UPGMA algorithm are the most suitable for cluster analysis. Based on the obtained data, the range of similarity was found in the range of 0.376 to 0.880 with an average of 0.626. In the cluster analysis, the samples were divided into two groups at the similarity level of 0.56, and the first group is the largest group, which can be divided into two subgroups. In the population structure analysis, the indicators of Observed number of alleles, Effective number of alleles, Nei's gene diversity and Shannon's Information index in populations and total population were calculated as 1.928, 1.641, 0.364 and 0.532, respectively. The total diversity, average intrapopulation diversity, level of population subdivision were also obtained as 0.380, 0.330 and 0.130 respectively. The maximum similarity was between the samples attributed to Darjeeling and selected Iranian samples.

Conclusion: Based on the obtained data, it was determined that there are significant changes among the investigated samples based on the ISSR markers. The percentage of polymorphism and polymorphism information content of the used markers indicated that these markers have a high ability to identify diversity among tea genotypes. Also, the results of this study stated that tea genotypes in Iran have high genetic diversity because they are mostly sexually propagated. This level of variation found between samples suggests that tea from different companies or regions is of better quality, not entirely due to genotype, but rather the ecology in which the tea is grown in terms of climate and soil characteristics, as well as processing technology. At the same time, this genetic difference can be used in breeding studies.

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1.Benzie, I. F., & Szeto, Y. T. (1999). Total antioxidant capacity of teas by the ferric reducing/antioxidant power assay. Journal of agricultural and food chemistry, 47 (2), 633-636.2.Ferruzzi, M. G. (2010). The influence of beverage composition on delivery of phenolic compounds from coffee and tea. Physiology & behavior, 100 (1), 33-41.3.Panda, H. (2011). The Complete Book on Cultivation and Manufacture of Tea. Asia Pacific Business Press, New Delhi.4.De Costa, W. A., Mohotti, A. J., & Wijeratne, M. A. (2007). Ecophysiology of tea. Brazilian Journal of Plant Physiology, 19, 299-332.5.Yogurtcu, B., & Aygun, A. (2021). Characterization of tea (Camellia sinensis L.) genotypes grown in Turkey by ISSR markers. Applied Ecology & Environmental Research, 19 (5), 10.15666/aeer/1905_41034114.6.Bublyk, O., Andreev, I., Kalendar, R., Spiridonova, K., & Kunakh, V. (2013). Efficiency of different PCR-based marker systems for assessment of Iris pumila genetic diversity. Biologia, 68 (4), 613-620.7.Anderson, N. O., Younis, A., & Sun, Y. (2010). Intersimple sequence repeats distinguish genetic differences in Easter lily ‘Nellie White’clonal ramets within and among bulb growers over years. Journal of the American Society for Horticultural Science, 135 (5), 445-455.8.Vo, T. D. (2007). Assessing genetic diversity in Vietnam tea [Camellia sinensis (L.) O. Kuntze] using morphology, inter-simple sequence repeat (ISSR) and microsatellite (SSR) markers. Dissertation, Georg-August University. http://dx.doi.org/10.53846/goediss-1693.9.Kalpana, D., Choi, S. H., Choi, T. K., Senthil, K., & Lee, Y. S. (2012). Assessment of genetic diversity among varieties of mulberry using RAPD and ISSR fingerprinting. Scientia Horticulturae, 134, 79-87.10.Kumar Mondal, T. (2002). Assessment of genetic diversity of tea (Camellia sinensis (L.) O. Kuntze) by inter-simple sequence repeat polymerase chain reaction. Euphytica, 128 (3), 307-315.11.Nybom, H. (2004). Comparison of different nuclear DNA markers for estimating intraspecific genetic
diversity in plants. Molecular ecology, 13 (5), 1143-1155.12.Thomas, J., Vijayan, D., Joshi, S. D., Lopez, S. J., & Kumar, R. R. (2006). Genetic integrity of somaclonal variants in tea (Camellia sinensis (L.) O Kuntze) as revealed by inter simple sequence repeats. Journal of Biotechnology, 123 (2), 149-154.13.Zietkiewicz, E., Rafalski, A., & Labuda, D. (1994). Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification. Genomics, 20 (2), 176-183.14.Chen, D., Yi, X., Yang, H., Zhou, H., Yu, Y., Tian, Y., & Lu, X. (2015). Genetic diversity evaluation of winged bean (Psophocarpus tetragonolobus (L.) DC.) using inter-simple sequence repeat (ISSR). Genetic Resources and Crop Evolution, 62, 823-828.15.Ben-Ying, L. I. U., You-Yong, L. I., Yi-Chun, T. A. N. G., Li-Yuan, W. A. N. G., Cheng, H., & Ping-Sheng, W. A. N. G. (2010). Assessment of genetic diversity and relationship of tea germplasm in Yunnan as revealed by ISSR markers. Acta Agronomica Sinica, 36 (3), 391-400.16.Chen, L., Zhou, Z. X., & Yang, Y. J. (2007). Genetic improvement and breeding of tea plant (Camellia sinensis) in China: from individual selection to hybridization and molecular breeding. Euphytica, 154, 239-248.17.Falakro, K., & Jahangirzadeh Khiavi, S. (2020). Assessment of genetic diversity and relationships among tea genotypes in Iran based on RAPD and ISSR markers. Journal of Horticulture and Postharvest Research, 3 (2), 209-220.18.Jahangirzadeh Khiavi, S., Falakro, K., Safaei Chaei Kar, S., Ramzi, S., & Kahneh, E. (2020). Usage of morphological and ISSR markers for investigation of Tea genotypes. Journal of Plant Production Research, 26 (4), 131-147. In Persian]19.Roy, S. C., & Chakraborty, B. N. (2009). Genetic diversity and relationships among tea (Camellia sinensis) cultivars as revealed by RAPD and ISSR based fingerprinting. Indian Journal of Biotechnology, 8 (4), 370-376.20.Yao, M. Z., Chen, L., & Liang, Y. R. (2008). Genetic diversity among tea cultivars from China, Japan and Kenya revealed by ISSR markers and its implication for parental selection in tea breeding programmes. Plant Breeding, 127 (2), 166-172.21.Beris, F. S., Sandalli, C., Canakci, S., Demirbag, Z., & Belduz, A. O. (2005). Phylogenetic analysis of tea clones (Camellia sinensis) using RAPD markers. Biologia-Section Botany, 60, 457-461.22.Dellaporta, S. L., Wood, J., & Hicks, J. B. (1983). A plant DNA minipreparation: version II. Plant molecular biology reporter, 1, 19-21.23.Roldan-Ruiz, I., Calsyn, E., Gilliland, T. J., Coll, R., Van Eijk, M. J. T., & De Loose, M. (2000). Estimating genetic conformity between related ryegrass (Lolium) varieties. 2. AFLP characterization. Molecular Breeding, 6, 593-602.24.Yun-Qing, H., Xiao-Bo, L., Xiao-Ling, W. (2019). Genetic diversity of the endangered Acer pentaphyllum Diels by ISSR analysis. Journal of Sichuan University (Natural Science Edition),
56 (1), 161-166.25.Kafkas, S., Ozkan, H., Ak, B., Acar, I., Atli, H., & Koyuncu, S. (2006). Detecting DNA polymorphism and genetic diversity in a wide pistachio germplasm: Comparison of AFLP, ISSR, and RAPD markers. Journal of the American Society for Horticultural Science, 131 (4), 10.21273/jashs.131. 4.522.26.Najafzadeh, R., Arzani, K., Bouzari, N., & Saei, A. (2014). Genetic diversity assessment and identification of new sour cherry genotypes using intersimple sequence repeat markers. International Journal of Biodiversity, 1-8. http://dx.doi.org/10.1155/2014/308398.27.Kumar, J., & Agrawal, V. (2019). Assessment of genetic diversity, population structure and sex identification in dioecious crop, Trichosanthes dioica employing ISSR, SCoT and SRAP markers. Heliyon, 5 (3), e01346.28.Noroozi, S., Baghızadeh, A., & Javaran, M. J. (2009). The genetic diversity of Iranian pistachio Pistacia vera L. cultivars revealed by ISSR markers. Biological Diversity and Conservation, 2 (2), 50-56.29.Indu, R., Kaushik, R. A., Deepak, R., Maloo, S. R., & Manish, C. (2020). Molecular characterization of chrysanthemum by using RAPD and ISSR markers. Journal of Pharmacognosy and Phytochemistry,
9 (1), 1423-1429. 30.Esselman, E. J., Jianqiang, L., Crawford, D. J., Windus, J. L., & Wolfe, A. D. (1999). Clonal diversity in the rare Calamagrostis porteri ssp. insperata (Poaceae): comparative results for allozymes and random amplified polymorphic DNA (RAPD) and intersimple sequence repeat (ISSR) markers. Molecular Ecology, 8 (3), 443-451.31.Nagaoka, T., & Ogihara, Y. (1997). Applicability of inter-simple sequence repeat polymorphisms in wheat for use as DNA markers in comparison to RFLP and RAPD markers. Theoretical and applied genetics, 94, 597-602.32.Paul, S., Wachira, F. N., Powell, W., & Waugh, R. (1997). Diversity and genetic differentiation among populations of Indian and Kenyan tea (Camellia sinensis (L.) O. Kuntze) revealed by AFLP markers. Theoretical and Applied Genetics, 94, 255-263.33.Yang, W., de Oliveira, A. C., Godwin, I., Schertz, K., & Bennetzen, J. L. (1996). Comparison of DNA marker technologies in characterizing plant genome diversity: variability in Chinese sorghums. Crop Science, 36 (6), 1669-1676.34.Jahangirzadeh Khiavi, S., & Falakro, K. (2017). Investigation of Genetic Diversity between Tea Shrubs Based on ISSR Markers. In Proceedings of First National Conference of Ecology, Diversity and Plant Conservation. Tehran, Iran. [In Persian]
35.Falakro, K., Khiavi, S. J., & Chaeikar, S. S. (2020). Study of Genetic Diversity between Some Tea Genotypes from Foman-Iran. Bioagrica, 1 (1), 1-10.36.Falakro, K., Jahangirzadeh Khiavi, S., Gholami, M., & Pour Azizan, S. (2022). RAPD markers, instruments for determining the genetic relationships of tea plant in Iran. Journal of Plant Research, 35 (1), 42-54.37.Kaundun, S. S., Zhyvoloup, A., & Park, Y. G. (2000). Evaluation of the genetic diversity among elite tea (Camellia sinensis var. sinensis) accessions using RAPD markers. Euphytica, 115, 7-16.38.Lai, J. A., Yang, W. C., & Hsiao, J. Y. (2001). An assessment of genetic relationships in cultivated tea clones and native wild tea in Taiwan using RAPD and ISSR markers. Botanical Bulletin of Academia Sinica, 42.39.Balasaravanan, T., Pius, P. K., Kumar, R. R., Muraleedharan, N., & Shasany, A. K. (2003). Genetic diversity among south Indian tea germplasm (Camellia sinensis, C. assamica and C. assamica spp. lasiocalyx) using AFLP markers. Plant Science, 165 (2), 365-372.40.Taniguchi, F., Kimura, K., Saba, T., Ogino, A., Yamaguchi, S., & Tanaka, J. (2014). Worldwide core collections of tea (Camellia sinensis) based on SSR markers. Tree genetics & genomes, 10, 1555-1565.