izvestiiatimacadИзвестия Тимирязевской сельскохозяйственной академииIZVESTIYA OF TIMIRYAZEV AGRICULTURAL ACADEMY0021-342XФГБОУ ВО РГАУ-МСХА имени К.А. Тимирязева10.26897/0021-342X-2022-6-43-53izvestiiatimacad-346Research ArticleГЕНЕТИКА, БИОТЕХНОЛОГИЯ, СЕЛЕКЦИЯ И СЕМЕНОВОДСТВОGENETICS, BIOTECHNOLOGY, BREEDING AND SEED PRODUCTIONФакторы прямого прорастания микроспорогенных эмбриоидов Brassica Napus L.Factors of direct germination of microspore derived embryos of Brassica napus LВишняковаА. В.VishnyakovaA. V.

Вишнякова Анастасия Васильевна, канд. с.-х. наук, доцент кафедры ботаники, селекции и семеноводства садовых растений

127550, г. Москва, ул. Тимирязевская, 49

тел.: (499) 976–41–71

Anastasiya V. Vishnyakova, PhD (Ag), Associate Professor of the Department of Botany, Plant Breeding and Seed Technology

49 Timiryazevskaya Str., Moscow, 127434

phone: (499) 976–41–71

a.vishnyakova@rgau-msha.ruАлександроваА. А.AleksandrovaA. A.

Александрова Анастасия Алексеевна, аспирант кафедры ботаники, селекции и семеноводства садовых растений

127550, Москва, ул. Тимирязевская, 49

тел.: (910) 466–03–09

Anastasiya A. Aleksandrova, post-graduate student of the Department of Botany, Plant Breeding and Seed Technology

49 Timiryazevskaya Str., Moscow, 127434

phone: (910) 466–03–09

a.alexandrova@rgau-msha.ruМонахосС. Г.MonakhosS. G.

Монахос Сократ Григорьевич, д-р с.-х. наук, профессор, заведующий кафедрой ботаники, селекции и семеноводства садовых растений

127550, г. Москва, ул. Тимирязевская, 49

тел.: (499) 976–41–71

Sokrat G. Monakhos, DSc (Ag), Professor, Head of the Department of Botany, Plant Breeding and Seed Technology

49 Timiryazevskaya Str., Moscow, 127434

phone: (499) 976–41–71

s.monakhos@rgau-msha.ru(Российский государственный аграрный университет – МСХА имени К.А. ТимирязеваРоссияRussian State Agrarian University – Moscow Timiryazev Agricultural AcademyRussian Federation202231032023164353Copyright © Вишнякова А.В., Александрова А.А., Монахос С.Г., 20232023Вишнякова А.В., Александрова А.А., Монахос С.Г.Vishnyakova A.V., Aleksandrova A.A., Monakhos S.G.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://izvestiia.timacad.ru/jour/article/view/346

Культура изолированных микроспор – основной метод производства удвоенных гаплоидов рапса, широко используемый в научно-исследовательских учреждениях и коммерческих компаниях. Протокол производства эмбриоидов рапса хорошо разработан и эффективен для многих генотипов, при этом сохраняются сложности, выражающиеся в низкой частоте регенерации проростков из эмбриоидов. При применении стандартного протокола регенерация проростков из микроспорогенных эмбриоидов, как правило, включает стадию каллусообразования и последующей регенерации адвентивных побегов, или вторичных эмбриоидов, что увеличивает срок производства растений-регенерантов и затрудняет производство удвоенных гаплоидов. Выявление факторов, влияющих на частоту прямого прорастания эмбриоидов, позволит увеличить частоту формирования проростков и сократить срок производства растений-регенерантов. При проведении исследований было изучено влияние кислотности среды на продолжительность регенерации проростков при культивировании микроспорогенных эмбриоидов рапса, а также влияние воздействия на микроспорогенные эмбриоиды в семядольной стадии развития пониженных температур +1 и +5℃ в течение 3, 6, 8, 9, 12 дней при культивировании в темноте на частоту прямого прорастания эмбриоидов. Повышение рН питательной среды с 5,8 до 6,1 увеличило частоту прямого прорастания эмбриоидов на 18% и общую частоту регенерации проростков с 46 до 76%. Культивирование эмбриоидов при низких положительных температурах повлияло на частоту прямого прорастания эмбриоидов в сеянцы: наблюдали максимальную частоту прямого прорастания эмбриоидов 44–53% при культивировании при +1℃ в течение 6 и 9 дней. При культивировании эмбриоидов при +5℃ частота прямого прорастания составляла 0–10%, в контрольном варианте без холодовой обработки – 16%.

Isolated microspore culture is the main method of producing doubled rapeseed haploids and is widely used in research institutions and commercial companies. The protocol of rapeseed embryo production is well developed and efficient for many genotypes, but some issues remain due to the low regeneration frequency of plantlets from embryos. When the standard protocol is applied, regeneration of plantlets from microspore-derived embryos usually involves a callus-forming stage 51 followed by regeneration of adventitious shoots or secondary embryos, which prolong the period of plantlet regeneration and makes production of doubled haploids complicated. Identifying factors, which affect the frequency of direct embryo germination will increase the frequency of plantlet formation and reduce the period of DH plant production. In this work, we studied the effect of medium pH on the duration of plantlet regeneration from rapeseed microspore-derived embryos and effect of their low temperature treatment of +1 and +5℃ for 3, 6, 8, 9, 12 days in complete darkness on the embryo maturation and germination. Rising the pH of the nutrient medium from 5.8 to 6.1 increased the frequency of direct embryos germination up to 18% and the overall frequency of plantlet regeneration up to 76%. Culturing embryos at low temperatures effected the frequency of direct germination of embryos into plantlets. The maximum frequency of 44–53% direct embryo germination was observed when cultured at +1℃ for 6 and 9 days, when embryos were cultured at +5℃ the frequency of direct germination was 0–10%. In the control variant without cold treatment it was 16%.

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