Clonal micropropagation of a rare species Hedysarum theinum Krasnob. (Fabaceae) and assessment of the genetic stability of regenerated plants using ISSR markers
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In the present study, a protocol was developed for the in vitro propagation of a rare medicinal plant, Hedysarum theinum (tea sweetvetch), from axillary buds, and identification of the regenerants was performed with the use of ISSR markers. It was demonstrated that Gamborg and Eveleigh medium supplemented with 5 μM 6-benzylaminopurine was the best for H. theinum for initial multiplication. On the other hand, half-strength Murashige and Skoog (MS) basal medium supplemented with 7 μM α-naphthaleneacetic acid proved to be the best for explant rooting. Molecular genetic analysis of the H. theinum mother plants and the obtained regenerants was performed with six ISSR markers. Depending on the primer, four to ten amplified fragments with sizes ranging from 250 to 3000 bp were identified. Our results confirmed the genetic stability of regenerants obtained in five passages and their identity to the mother plant.
KeywordsInter Simple Sequence Repeat Genetic Stability Naphthalene Acetic Acid ISSR Marker Genetic Fidelity
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- 1.Krasnoborov, I.M., Azovtsev, G.R., and Orlov, V.P., A new species of the genus Hedysarum L. (Fabaceae L.) from southern Siberia, Bot. Zh., 1985, vol. 70, no. 7, pp. 968–973.Google Scholar
- 2.Kurbatskii, V.I., Genus Hedysarum L., in Flora Sibiri (Flora of Siberia), Polozhii, A.V., et al., Eds., Novosibirsk: Nauka, 1994, vol. 9, pp. 153–166.Google Scholar
- 3.Volodarskaya, S.B., Vinokurova, E.Yu., and Shul’ts, E.E., Chemical study of Hedysarum theinum, in Fizicheskie i biokhimicheskie aspekty izucheniya lekarstvennykh rastenii (Physical and Biochemical Aspects of Medicinal Plants’ Investigation) (Proc. Int. Conf. in Memoriam of V.G. Minaev), Novosibirsk, 1998, pp. 18–19.Google Scholar
- 21.Hakki, E.E., Dogan, B., Duran, A., et al., Phylogenetic relationship analysis of Genista L. (Fabaceae) species from Turkey as revealed by inter-simple sequence repeat amplification, Afr. J. Biotechnol., 2010, vol. 9, no. 18, pp. 2627–2632.Google Scholar
- 22.Vysochina, G.I. and Kukushkina, T.A., Biologically active substances from some Hedysarum L. species, Khim. Rastit. Syr’ya, 2011, vol. 4, pp. 251–258.Google Scholar
- 26.Elisafenko, T.V., Dorogina, O.V., Achimova, A.A., and Yamtyrov, M.B., Problems of reintroduction and restoration by example of species from the genera Hedysarum L. and Viola L., in Problemy botaniki Yuzhnoi Sibiri i Mongolii (Problems of Botany of Southern Siberia and Mongolia) (Proc. 12th Int. Theor. Pract. Conf.), Barnaul, 2013, pp. 232–234.Google Scholar
- 27.Vysotskii, V.A., Biotechnological methods in the production of improved planting material and selection of fruit and berry plants, Extended Abstract of Doctoral Dissertation, Moscow, 1998.Google Scholar
- 28.De Klerk, G.J., How to measure somaclonal variation, Acta Bot. Neerl., 1990, vol. 39, pp. 129–144.Google Scholar
- 29.Sharma, S.K., Bryan, G.J., Winfield, M.O., and Millam, S., Stability of potato (Solanum tuberosum L.) plants regenerated via somatic embryos, axillary bud proliferated shoots, microtubers and true potato seeds: a comparative phenotypic, cytogenetic and molecular assessment, Planta, 2007, vol. 226, no. 6, pp. 1449–1458.CrossRefPubMedGoogle Scholar
- 32.Nayak, S. and Sen, S., Differential resistance of three species of Ornithogalum to polyploidization in vitro, Nucleus, 1998, vol. 41, pp. 48–52.Google Scholar