vavilovВавиловский журнал генетики и селекцииVavilov Journal of Genetics and Breeding2500-3259Institute of Cytology and Genetics of Siberian Branch of the RAS10.18699/VJ17.272vavilov-1139Research ArticleГЕНОМИКА И ТРАНСКРИПТОМИКАPLANT GENETICS AND BREEDINGСохранение эндемика Западного Саяна Fritillaria sonnikovae Schaulo et A. Erst (Liliaceae) в коллекции in vitroConservation of the West Sayan endemic Fritillaria sonnikovae Schaulo et A. Erst (Liliaceae) in in vitro collectionМурасеваД. С.MurasevaD. S.

Новосибирск

Novosibirsk

dsmuraseva@csbg.nsc.ruЗвягинаН. С.ZvyaginaN. S.

Новосибирск

Novosibirsk

НовиковаТ. И.NovikovaT. I.

Новосибирск

Novosibirsk

ДорогинаО. В.DoroginaO. V.

Новосибирск

Novosibirsk

Центральный сибирский ботанический сад Сибирского отделения Российской академии наукРоссияCentral Siberian Botanical Garden SB RASRussian Federation201701112017215554560Copyright © Мурасева Д.С., Звягина Н.С., Новикова Т.И., Дорогина О.В., 20172017Мурасева Д.С., Звягина Н.С., Новикова Т.И., Дорогина О.В.Muraseva D.S., Zvyagina N.S., Novikova T.I., Dorogina O.V.This work is licensed under a Creative Commons Attribution 4.0 License.https://vavilov.elpub.ru/jour/article/view/1139

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.).

An effective system of micropropagation and in vitro conservation of the West Sayan endemic Fritillaria sonnikovae Schaulo et A. Erst was developed and the genetic stability of regenerants after slow-growth storage was assessed. The efficiency of bulb scale segments as primary explants was established. The optimum nutrient medium for stages of in vitro culture initiation and multiplication was BDS supplemented with BAP (5.0 µM) and NAA (2.0 µM). The microplant collection was maintained in active-growth (+23±2 °С) and slow-growth (+7 °С) conditions. The period of subculturing at +7 °С was prolonged to 9–12 months when microclones of F. sonnikovae were transferred to slowgrowth conditions. Assessment of microplant regeneration potential in further cultivation of bulb scales on BDS nutrient medium supplemented with BAP (5.0 µM) and NAA (2.0 µM) revealed a stimulating effect of conservation at low temperatures: the regeneration rate reached 93 % and the number of bulblets per explant was 6.9±1.7 at the first passage after slow-growth storage. However, further cultivation led to decrease of shoot development, and the regeneration rate reached the level before slow-growth storage at the third passage. The genetic fidelity of regenerants obtained during direct organogenesis at the first passage after slow-growth storage (12 months) was established by analysis of ISSR-PCR-fragments.

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The authors declare that there are no conflicts of interest present.