ovoshchiОвощи РоссииVegetable crops of Russia2072-91462618-7132Федеральный научный центр овощеводства10.18619/2072-9146-2022-5-5-14ovoshchi-2026Research ArticleСЕЛЕКЦИЯ, СЕМЕНОВОДСТВО И БИОТЕХНОЛОГИЯ РАСТЕНИЙBREEDING, SEED PRODUCTION AND PLANT BIOTECHNOLOGYОптимизация этапов технологии получения удвоенных гаплоидов кабачка (Cucurbita pepo L.) в культуре неопыленных семяпочек in vitroOptimization of steps in the technology of obtaining doubled haploids of summer squash (Cucurbita pepo L.) in the culture of unpollinated ovules in vitrohttps://orcid.org/0000-0002-7433-5271ЕрмолаевА. С.ErmolaevA. S.

Алексей Станиславович Ермолаев – младший научный сотрудник лаборатории репродуктивной биотехнологии в селекции сельскохозяйственных растений

143072, Московская область, Одинцовский район, п. ВНИИССОК, ул. Селекционная, д.14

Alexey S. Ermolaev – Junior Researcher of Laboratory of Reproductive Biotechnology in Crop Breeding 

14, Selectsionnaya str., VNIISSOK, Odintsovo district, Moscow region, 143072

ErmolaevAlexeyStanislavovich@gmail.comhttps://orcid.org/0000-0002-2695-190XДомблидесЕ. А.DomblidesE. A.

Елена Алексеевна Домблидес – кандидат с.-х. наук, зав. лаб. репродуктивной биотехнологии в селекции с.-х. растений

143072, Московская область, Одинцовский район, п. ВНИИССОК, ул. Селекционная, д.14

Elena A. Domblides – Cand. Sci. (Agriculture), Head of Laboratory of Reproductive Biotechnology in Crop Breeding 

14, Selectsionnaya str., VNIISSOK, Odintsovo district, Moscow region, 143072

edomblides@mail.ruФедеральное государственное бюджетное научное учреждение "Федеральный научный центр овощеводства" (ФГБНУ ФНЦО)РоссияFederal State Budgetary Scientific Institution Federal Scientific Vegetable Center (FSBSI FSVC)Russian Federation20222509202205514Copyright © Ермолаев А.С., Домблидес Е.А., 20222022Ермолаев А.С., Домблидес Е.А.Ermolaev A.S., Domblides E.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.vegetables.su/jour/article/view/2026

Актуальность. Для создания эффективной технологии получения удвоенных гаплоидов (DH-технологии) кабачка в культуре неопыленных семяпочек in vitro необходимо подобрать оптимальные значений многих факторов, степень влияния каждого из которых на гиногенез может существенно отличаться. Целью исследования являлась оптимизация отдельных этапов технологии.Методы. Для индукции эмбриогенеза использовали жидкую и агаризованную (7 г/л) среду IMC с различной концентрацией сахарозы (от 20 до 80 г/л) и различными регуляторами роста растений (2 мг/л 2,4 D; 0,2 мг/л ТДЗ; 0,8 мг/л 2,4 D и 1,2 мг/л НУК).Результаты. Оптимальным для изученных генотипов кабачка оказался предварительно заизолированный с вечера, сорванный утром раскрывшийся бутон. Стерилизация завязей кабачка краткосрочным обжиганием после обработки 96% спиртом, позволяет существенно сократить временные затраты на этот этап без потери эмбриогенного потенциала. Для индукции гиногенеза в культуре неопыленных семяпочек кабачка возможно использовать жидкую питательную среду IMC c сахарозой от 20 до 40 г/л. Использование питательной среды с 2 мг/л 2,4 D для большинства генотипов оказалось более эффективным и позволяло получить большее количество эмбриоидов, по сравнению с питательной средой содержащей 0,2 мг/л ТДЗ и со средой с 0,8 мг/л 2,4 D и 1,2 мг/л НУК. Образование эмбриоидов наблюдалось уже через 5 недель культивирования.Заключение. Нам удалось завершить полный цикл технологии получения удвоенных гаплоидов в культуре неопыленных семяпочек in vitro для 30 генотипов кабачка и получить DH-растения, которые являются ценным исходным материалом как для селекционеров, так и для генетических исследований. Оптимизация этапов технологии позволила достичь максимального результата для отдельных генотипов – 55 эмбриоидов на 100 культивируемых семяпочек.

Relevance. To create an effective technology for obtaining doubled haploids (DH-technology) of zucchini in unpollinatedseedpod culture in vitro it is necessary to select the optimal values of many factors, the degree of influence of each of which on gynogenesis can vary significantly. The aim of the study was to optimize the individual stages of the technology.Methods. Liquid and agarized (7 g/L) IMC medium with different sucrose concentrations (20 to 80 g/L) and different plant growth regulators (2 mg/L 2,4 D; 0.2 mg/L TDZ ; 0.8 mg/L 2,4 D and 1.2 mg/L NUC) were used for induction of embryogenesis.Results. Optimal for the studied zucchini genotypes was pre-isolated from the evening, plucked in the morning opened bud. Sterilization of zucchini ovaries by short-term burning after treatment with 96% alcohol, allows significant reduction of the time required for this step without loss of embryogenic potential. IMC nutrient medium with sucrose (20 to 40 g/l) can be used for induction of gynogenesis in the unpollinatedzucchini ovary culture. The use of nutrient media with 2 mg/l 2,4 D for most genotypes was more effective and resulted in higher number of embryoids compared to nutrient media containing 0.2 mg/l TDC and media with 0.8 mg/l 2,4 D and 1.2 mg/l NAA. Embryoid formation was observed after 5 weeks of cultivation.Conclusion. We were able to complete the full cycle of technology for obtaining doubled haploids in unpollinatedseedpod culture in vitro for 30 zucchini genotypes and obtain DHplants, which are valuable source material for both breeders and genetic research. Optimization of the individual steps of the technology made it possible to achieve the maximum result for individual genotypes – 55 embryoids per 100 cultivated ovules.

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