vavilovВавиловский журнал генетики и селекцииVavilov Journal of Genetics and Breeding2500-3259Institute of Cytology and Genetics of Siberian Branch of the RAS10.18699/VJ20.592vavilov-2477Research ArticleБИОТЕХНОЛОГИЯ И СЕЛЕКЦИЯ РАСТЕНИЙBIOTECHNOLOGY AND PLANT BREEDINGФакторы, влияющие на получение DH-растений в культуре микроспор in vitro редиса европейскогоFactors affecting DH plants in vitro production from microspores of European radishhttps://orcid.org/0000-0001-5447-5341КозарьЕ. В.KozarE. V.

Пос. ВНИИССОК, Одинцовский район, Московская область

VNIISSOK, Odintsovo region, Moscow oblast

koz.leno4ek@gmail.comhttps://orcid.org/0000-0002-2695-190XДомблидесЕ. А.DomblidesE. A.

Пос. ВНИИССОК, Одинцовский район, Московская область

VNIISSOK, Odintsovo region, Moscow oblast

edomblides@mail.ruhttps://orcid.org/0000-0002-9492-6845СолдатенкоА. В.SoldatenkoA. V.

Пос. ВНИИССОК, Одинцовский район, Московская область

VNIISSOK, Odintsovo region, Moscow oblast

Федеральное государственное бюджетное научное учреждение Федеральный научный центр овощеводстваРоссияFederal State Budgetary Scientific Institution Federal Scientific Vegetable CenterRussian Federation2020170320202413139Copyright © Козарь Е.В., Домблидес Е.А., Солдатенко А.В., 20202020Козарь Е.В., Домблидес Е.А., Солдатенко А.В.Kozar E.V., Domblides E.A., Soldatenko A.V.This work is licensed under a Creative Commons Attribution 4.0 License.https://vavilov.elpub.ru/jour/article/view/2477

В последние годы резко повысилась потребность рынка в увеличении производства сортов и гибридов редиса европейского (RaphanussativusL.) для открытого и закрытого грунта, разнообразных по группам спелости, форме и окраске корнеплода. Поэтому важно расширять генетическое разнообразие и ускорять селекционный процесс. Технология получения удвоенных гаплоидов существенно сокращает время при создании гомозиготных константных родительских линий, для получения которых наиболее перспективен метод культуры микроспор invitro. Нам впервые удалось осуществить полный цикл получения DH-растений редиса европейского в культуре микроспор invitro, до включения материала в селекционный процесс. Подобраны: оптимальный размер бутонов, параметры теплового шока, среды для индукции и регенерации. Выявлено, что линейная длина бутонов с оптимальной стадией развития микроспор генотип-специфична. Так, для сортообразца Родос оптимальным является показатель 2.8-3.3 мм, а для сортообразца Тепличный Грибовский -3.7-4.2 мм. Для большинства генотипов оптимален температурный шок 32 °С в течение 48 ч. Впервые для индукции эмбриогенеза использована модифицированная среда Мурасиге-Скуга и обнаружено существенное влияние взаимодействия факторов «генотип x среда» на индукцию эмбриогенеза. Для этапа регенерации растений из эмбриоидов рекомендуется добавление к среде 1 мг/л бензиламинопурина и 0.1 мг/л гиббереллино-вой кислоты, укоренение микропобегов проводится на безгормональной среде. Анализ полученных растений-регенерантов методом подсчета хромосом и методом проточной цитометрии клеточных ядер показал, что 69 % растений имели диплоидный набор хромосом, 9 % - гаплоидный, 22 % - миксоплоидный и анеуплоидный. Семенное потомство удалось получить самоопылением из удвоенных гаплоидов и миксоплоидов, причем все растения R1 имели удвоенный набор хромосом. Это исследование является начальным этапом в разработке эффективной методики получения удвоенных гаплоидов редиса для применения в селекционном процессе.

Over the recent years the market demand for scaling up the production of European radish (Raphanus sativus L.) varieties and hybrids for open and protected production, varying in ripeness group, root shape and color, has drastically increased. Therefore, the expansion of genetic diversity and acceleration of the selection process are important. Doubled haploid technology considerably curtails the time required for creation of homozygous constant parental cell lines when in vitro microspore culture is used as the most promising method. For the first time, we were able to realize the full production cycle of DH plants of European radish by in vitro microspore culture up to inclusion of the produced material into the selection process. We have selected: preferable flower bud size, heat shock parameters, induction and regeneration media. It was revealed that linear length on the flower buds with the best possible stage of microspore development is genotype-specific: the flower bud length 2.8-3.3 mm is optimal for accessions of Rhodes and 3.7-4.2 mm is optimal for accessions of Teplichny Gribovsky. Heat shock at 32 °C for 48 hours is the most suitable for most genotypes. For the first time Murashige and Skoog based culture medium has been used for embryogenesis induction, and a major dependence of embryogenesis induction on the genotype x medium interaction was found. At regeneration and tiller stage it is advisable to add 1 mg/mL of benzylaminopurine and 0.1 mg/L of gibberellic acid to the medium, and rotting of micro-sprouts is performed with the use of hormone-free medium. Analysis of the produced regenerant plants by chromosome count and cell nucleus flow cytometry showed that 69 % of plants have a diploid chromosome set, 9 % have a haploid chromosome set, and 22 % have mixoploids and aneu-ploids chromosome sets. The seed progeny from doubled haploids and mixoploids were obtained by self-pollination, where all R1 plants had a doubled set of chromosomes. This study launches the development of an efficient method of radish doubled haploid production to be used in the selection process.

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