vavilovВавиловский журнал генетики и селекцииVavilov Journal of Genetics and Breeding2500-3259Institute of Cytology and Genetics of Siberian Branch of the RAS10.18699/vjgb-25-55vavilov-4677Research ArticleГЕНЕТИКА РАСТЕНИЙPLANT GENETICSОптимизация этапов технологии получения DH-растений капусты белокочаннойOptimization of technology steps for obtaining white cabbage DH-plantsМинейкинаА. И.MineykinaA. I.

пос. ВНИИССОК, Одинцовский район, Московская о6ласть

VNIISSOK, Odintsovo district, Moscow region

СтебницкаяК. С.StebnitskaiaK. S.

пос. ВНИИССОК, Одинцовский район, Московская о6ласть

VNIISSOK, Odintsovo district, Moscow region

ФомичеваМ. Г.FomichevaM. G.

пос. ВНИИССОК, Одинцовский район, Московская о6ласть

VNIISSOK, Odintsovo district, Moscow region

БондареваЛ. Л.Bondareva,L. L.

пос. ВНИИССОК, Одинцовский район, Московская о6ласть

VNIISSOK, Odintsovo district, Moscow region

ДомблидесА. С.DomblidesA. S.

пос. ВНИИССОК, Одинцовский район, Московская о6ласть

VNIISSOK, Odintsovo district, Moscow region

ДомблидесЕ. А.DomblidesE. A.

пос. ВНИИССОК, Одинцовский район, Московская о6ласть

VNIISSOK, Odintsovo district, Moscow region

edomblides@mail.ruФедеральный научный центр овощеводстваРоссияFederal Scientific Vegetable CenterRussian Federation202520072025294517529Copyright © Минейкина А.И., Стебницкая К.С., Фомичева М.Г., Бондарева Л.Л., Домблидес А.С., Домблидес Е.А., 20252025Минейкина А.И., Стебницкая К.С., Фомичева М.Г., Бондарева Л.Л., Домблидес А.С., Домблидес Е.А.Mineykina A.I., Stebnitskaia K.S., Fomicheva M.G., Bondareva, L.L., Domblides A.S., Domblides E.A.This work is licensed under a Creative Commons Attribution 4.0 License.https://vavilov.elpub.ru/jour/article/view/4677

Одна из экономически важных сельскохозяйственных культур среди представителей рода Brassica L. – капуста 6елокочанная. Для создания высокопродуктивных F1 ги6ридов с улучшенными показателями нео6ходима генетически разноо6разная 6аза выровненного материала, получение которого занимает продолжительную часть селекционного процесса. Ускорить этот этап селекции можно за счет получения удвоенных гаплоидов (DH-растений). Отсутствие стандартизированных, эффективных и воспроизводимых протоколов для культивирования in vitro разных видов растений, охватывающих несколько факторов и их взаимодействие, часто препятствует практической реализации метода. Растительный материал, условия культивирования и состав питательных сред являются определяющими факторами эффективности эм6риогенеза. В результате проведенного исследования оптимизирован протокол получения удвоенных гаплоидов в культуре изолированных микроспор in vitro для капусты 6елокочанной позднеспелого срока созревания. Оптимальный размер 6утонов позднеспелой капусты 6елокочанной для введения в культуру in vitro варьировал от 3.5 до 5.0 мм. Для изученных генотипов капусты 6елокочанной совместное влияние высокотемпературного стресса при 32 °С в течение 48 ч и рН питательной среды 5.8 спосо6ствовало наи6ольшему выходу эм6риоидов. 3а счет использования платформы-шейкера при режиме 40 о6./мин достигнуто ускорение развития эм6риоидов до семядольной стадии на 7–10 суток. Благодаря подо6ранному комплексу условий для успешного эм6риогенеза из микроспор капусты 6елокочанной достигнут выход эм6риоидов до 273.6 ± 32.2 шт./чашку Петри. Применение проточной цитометрии позволило отделить удвоенные гаплоиды (69.8 %) от гаплоидов (8.4 %), триплоидов (1.5 %) и тетраплоидов (20.3 %) на ранней стадии развития. Молекулярно-генетический анализ с использованием микросателлитных маркеров (SSR-анализ) подтвердил гаплоидное происхождение диплоидных растений-регенерантов.

White cabbage is one of the economically important crops among the representatives of the genus Brassica L. To create highly productive F1 hybrids with improved characteristics, the breeders need genetically diverse breeding material, which takes a long time to produce. It is possible to significantly accelerate this stage of breeding by obtaining doubled haploids (DH-plants). The lack of standardized, efficient and reproducible protocols for in vitro cultivation of different plant species, covering several factors and their interactions, often hinders the practical implementation of the method. Plant material, cultivation conditions and composition of nutrient media are determinants of embryogenesis efficiency. As a result of this study, the protocol for obtaining doubled haploids in in vitro culture of isolated microspores was optimized for late maturing white cabbage. The optimal bud size for introduction into in vitro culture varied from 3.5 to 5.0 mm. For the studied genotypes, the combined effect of high-temperature stress at 32 °C for 48 h and pH 5.8 stimulated the highest embryoid yield. The use of 3.5 g/L phytogel as a gelling agent was not effective. The use of flow cytometry allowed for separation of doubled haploids (69.8 %) from haploids (8.4 %), triploids (1.5 %) and tetraploids (20.3 %) at an early stage of development. Molecular genetic analysis with polymorphic microsatellite loci (SSR-analysis) confirmed the haploid origin of the diploid regenerant plants.

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