Сохранение эндемика Западного Саяна Fritillaria sonnikovae Schaulo et A. Erst (Liliaceae) в коллекции in vitro

Разработана эффективная система микроразмножения и сохранения в коллекции in vitro эндемика Западного Саяна Fritillaria sonnikovae Schaulo et A. Erst и проведена оценка генетической стабильности регенерантов после депонирования. Установлена эффективность использования сегментов луковичных чешуй в качестве первичных эксплантов. Наиболее оптимальной питательной средой на этапах инициации культуры in vitro и размножения являлась среда BDS, дополненная 5.0 мкМ 6-бензиламинопурина (БАП) и 2.0 мкМ 1-нафтилуксусной кислоты (НУК). Коллекция микрорастений поддерживается в условиях активного (+23±2 °С) и замедленного роста (+7 °С). При переводе микроклонов F. sonnikovae в условия замедленного роста при температуре +7 °С период беспересадочного субкультивирования удалось увеличить до 9–12 мес. Оценка регенерационного потенциала микрорастений при последующем культивировании сегментов чешуй на питательной среде BDS, дополненной 5.0 мкМ БАП и 2.0 мкМ НУК, позволила выявить стимулирующий эффект хранения при низких температурах: в первом пассаже после депонирования частота регенерации составила 93 %, количество адвентивных побегов – 6.9±1.7 шт./эксп. Однако дальнейшее субкультивирование приводило к снижению активности побегообразования, и к третьему пассажу регенерация достигала уровня до депонирования. На основании анализа ISSR-ПЦР-фрагментов установлено генетическое соответствие материнских растений и регенерантов, полученных в ходе прямого органогенеза в первом пассаже после депонирования (12 мес.).

Fritillaria sonnikovae, коллекция in vitro, ISSR-анализ, сомаклональная изменчивость, депонирование in vitro, сохранение биоразнообразия

1. Al Khateeb W., Bahar E., Lahham J., Schroeder D., Hussein E. Regeneration and assessment of genetic fidelity of the endangered tree Moringa peregrina (Forsk.) Fiori using Inter Simple Sequence Repeat (ISSR). Physiol. Mol. Biol. Plants. 2013;19(1):157-164. DOI 10.1007/ s12298-012-0149-z.

2. Antonova O.Yu., Truskinov E.V., Frolova D.V., Gavrilenko T.A. Analysis of the genetic stability of potato samples stored under in vitro conditions. Agrarnaya Rossiya = Agricultural Russia. 2004;6:25-30. (in Russian)

3. Bacchetta L., Remotti P.C., Bernardini C., Saccardo F. Adventitious shoot regeneration from leaf explants and stem nodes of Lilium. Plant. Cell Tiss. Org. Cult. 2003;74:37-44. DOI 10.1023/A:1023315321931.

4. Bairu M.W., Aremu A.O., Van Staden J. Somaclonal variation in plants: causes and detection methods. Plant Growth Regul. 2011;63:147-173. DOI 10.1007/s10725-010-9554-x.

5. Bairu M.W., Fennell C.W., Van Staden J. The effect of plant growth regulators on somaclonal variation in Cavendish banana (Musa AAA cv. “Zelig”). Sci. Hortic. 2006;108(4):347-351. DOI 10.1016/j.scienta.2006.01.039.

6. Bell R.L., Reed B.M. In vitro tissue culture of pear: Advances in techniques for micropropagation and germplasm preservation. Proc. 8th IS on Pear. 2002;412-418. Benson E.E. Plant Conservation Biotechnology. L.; Philadelphia P.A.: Taylor and Francis, 2002.

7. Bonnier F.J.M., Van Tuyl J.M. Long term in vitro storage of lily: effects of temperature and concentration of nutrients and sucrose. Plant Cell Tiss. Org. Cult. 1997;49:81-87. DOI 10.1023/A:1005810728215.

8. Bublyk O.M., Andreev I.O., Spiridonova K.V., Kunakh V.A. Genetic variability in regenerated plants of Ungernia victoris. Biologia Plantarum. 2012;56(2):395-400. DOI 10.1007/s10535-012-0106-2.

9. Cordeiro S.Z., Simas N.K., Henriques A.B., Sato A. In vitro conservation of Mandevilla moricandiana (Apocynaceae): short-term storage and encapsulation-dehydration of nodal segments. In Vitro Cell. Dev. Biol. – Plant. 2014;50:326-336. DOI 10.1007/s11627-014- 9600-x.

10. Cruz-Cruz C.A., González-Arnao M.T., Engelmann F. Biotechnology and conservation of plant biodiversity. Resources. 2013;2:73-95. DOI 10.3390/resources2020073.

11. Dunstan D.J., Short K.C. Improved growth of tissue cultures of the onion Allium cepa. Phisiol. Plant. 1977;41(1):70-72.

12. Engelmann F. Use of biotechnologies for the conservation of plant biodiversity. In vitro Cell. Dev. Biol. – Plant. 2011;47:5-16. DOI 10.1007/s11627-010-9327-2.

13. Gamborg O.L., Eveleigh D.E. Culture methods and detection of glucanases in cultures of wheat and barley. Can. J. Biochem. 1968;46(5): 417-421.

14. Jain S.M., Ochatt S.J. Protocols for in vitro propagation of ornamental plants, methods in molecular biology. New Jersey: Humana Press, 2010. DOI 10.1007/978-1-60327-114-1.

15. Jevremović S., Petrić M., Zivković S., Trifunović M., Subotić A. Superoxide dismutase activity and isoenzyme profiles in bulbs of snake’s head fritillary in response to cold treatment. Arch. Biol. Sci. 2010; 62(3):553-558. DOI 10.2298/ABS1003553J.

16. Joshi S.K., Dhar U., Andola H.C. In vitro bulblet regeneration and evaluation of Fritillaria roylei Hook. – a high value medicinal herb of the Himalaya. Acta Hortic. 2007;756:75-84. DOI 10.17660/ActaHortic.2007.756.8.

17. Kamenetsky R., Okubo H. Ornamental geophytes: from basic science to sustainable production. Boca Raton, F.L.: CRC Press, 2013.

18. Keller J. Influence of different temperature treatments on viability of in vitro cultivated Allium shoots and bulblets. Acta Hort. 1992;319:307-312. DOI 10.17660/ActaHortic.1992.319.47.

19. Krishnan P.N., Decruse S.W., Radha R.K. Conservation of medicinal plants of Western Ghats, India and its sustainable utilization through in vitro technology. In Vitro Cell. Dev. Biol. – Plant. 2011;47:110-122. DOI 10.1007/s11627-011-9344-9.

20. Kul’khanova D.S., Erst A.A., Novikova T.I. In vitro regeneration from bulbous scales of Fritillaria sonnikovae, an endemic species. Ontogenez = Ontogenesis (Moscow). 2015;46(4):259-266. DOI 10.7868/ S0475145015040059. (in Russian)

21. Liu X., Yang G. Adventitious shoot regeneration of oriental lily (Lilium orientalis) and genetic stability evaluation based on ISSR marker variation. In Vitro Cell. Dev. Biol.– Plant. 2012;48:172-179. DOI 10.1007/s11627-012-9429-0.

22. Marinangeli P.А., Hernandez L.F., Pellegrini C.P., Curvetto N.R. Bulblet differentiation after scale propagation of Lilium longiflorum. J. Am. Soc. Hort. Sci. 2003;128(3);324-329.

23. Miller M.P. Tools for populations genetic analyses (TFPGA) 1.3. A Windows program for the analysis of allozyme and molecular population genetic data. 1997. Available at http://www.ccg.unam. mx/~vinuesa/tlem09/docs/TFPGADOC.PDF.

24. Nei M. Genetic distance between populations. Am. Nat. 1972;106: 283-292.

25. Nikolić М., Misić D., Maksimović V., Jevremović S., Trifunović M., Subotić A. Effect of low temperature on rooting rate and carbohydrate content of Fritillaria meleagris bulbs formed in culture in vitro. Arch. Biol. Sci., Belgrade. 2008;60(1):5-6. DOI 10.2298/ ABS080105PN.

26. Novikova T.I. Use of biotechnological approaches for the conservation of plant biodiversity. Rastitelnyy mir Aziatskoy Rossii = Plant Life of Asian Russia. 2013;2(12):119-128. (in Russian)

27. Parić А., Cakar J., Muratovic E., Karalija E. Induction of bulblets on leaf and bulb explants of endangered Lilium bosniacum (G. Beck) G. Beck ex Fritsch. Botanica Serbica. 2011;35(1):31-35.

28. Poluboyarova T.V., Novikova T.I. Clonal micropropagation of ornamental onions of the subgenus Melanocrommyum from flower organs. Byulleten Gosudarstvennogo Nikitskogo botanicheskogo sada = Bulletin of the State Nikitskiy Botanical Gardens. 2013;107:46-52. (in Russian)

29. Rakosy-Tican E., Bors B., Szatmari A.-M. In vitro culture and mediumterm conservation of the rare wild species Gladiolus imbricatus. African J. Biotech. 2012;11(81):14703-14712. DOI 10.5897/AJB12.784.

30. Rani V., Parida A., Raina S. Random amplified polymorphic DNA (RAPD) markers for genetic analysis in micropropagated plants of Populus deltoides Marsh. Plant. Cell. Report. 1995;14:459-462. DOI 10.1007/BF00234055.

31. Reed B.M., Sarasan V., Kane M., Bunn E., Pence V.C. Biodiversity conservation and conservation biotechnology tools. In Vitro Cell Dev. Biol. – Plant. 2011;47:1-4. DOI 10.1007/s11627-010-9337-0.

32. Shaulo D.N., Erst A.S. A new species of Fritillaria L. (Liliaceae) from West Sayan. Turczaninowia. 2010;13(3):46-49. (in Russian)

33. Song J.Y., Mattson N.S., Jeong B.R. Efficiency of shoot regeneration from leaf, stem, petiole and petal explants of six cultivars of Chrysanthemum morifolium. Plant. Cell Tiss. Org. Cult. 2011;107:295- 304. DOI 10.1007/s11240-011-9980-0.

34. Tavartkiladze O.K., Vechernina N.A. Conservation of the plant gene pool in in vitro culture collections. Izvestiya Altayskogo gosudarstvennogo universiteta = Izvestiya of the Altai State University. 1999;5:13-15. (in Russian)

35. Varshney A., Lakshmikumaran M., Srivastava P.S., Dhawan V. Establishment of genetic fidelity of in vitro-raised Lilium bulblets through RAPD markers. In Vitro Cell. Dev. Biol. – Plant. 2001;37:227-231. DOI 10.1007/s11627-001-0040-z.

36. Vasil I.K., Thorpe T.A. Plant cell and tissue culture. Dordrecht: Kluwer Academic Publishers, 1994.

37. Vinci V.A., Parekh S.R. Handbook of industrial cell culture: mammalian, microbial and plant cells. New Jersey: Humana Press, 2003. DOI 10.1007/978-1-59259-346-0.

38. Zvyagina N.S., Dorogina O.V., Krasnikov A.A. Genetic differentiation and karyotype variation in Hedysarum chaiyrakanicum, an endemic species of Tuva Republic, Russia. Indian J. Experim. Biol. 2016; 54(5):338-344.