ovoshchiОвощи РоссииVegetable crops of Russia2072-91462618-7132Федеральный научный центр овощеводства10.18619/2072-9146-2021-4-11-26ovoshchi-1789Research ArticleСЕЛЕКЦИЯ И СЕМЕНОВОДСТВО СЕЛЬСКОХОЗЯЙСТВЕННЫХ РАСТЕНИЙBREEDING AND SEED PRODUCTION OF AGRICULTURAL CROPSПолучение удвоенных гаплоидов Cucurbita pepo L.Obtaining doubled haploids of Cucurbita pepo L.https://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, Россия, Московская область, Одинцовский район, п. ВНИИССОК, ул. Селекционная, д. 14

Elena A. Domblides

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

edomblides@mail.ruhttps://orcid.org/0000-0002-7433-5271ЕрмолаевЕ. А.ErmolaevA. S.

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

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

Alexey S.Ermolaev

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

AlexeyErmolaev@vniissok.ruhttps://orcid.org/0000-0002-4387-9153БеловС. Н.BelovS. N.

Сергей Николаевич Белов - младший научный сотрудник лаборатории репродуктивной биотехнологии в селекции сельскохозяйственных растений

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

Sergey N. Belov

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

belov@vniissok.ruФедеральное государственное бюджетное научное учреждение "Федеральный научный центр овощеводства" (ФГБНУ ФНЦО)РоссияFederal State Budgetary Scientific Institution Federal Scientific Vegetable Center (FSBSI FSVC)Russian Federation202102092021041126Copyright © Домблидес Е.А., Ермолаев Е.А., Белов С.Н., 20212021Домблидес Е.А., Ермолаев Е.А., Белов С.Н.Domblides E.A., Ermolaev A.S., Belov S.N.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.vegetables.su/jour/article/view/1789

Удвоенные гаплоиды уже около 100 лет используются во всем мире в селекционных программах и фундаментальных исследованиях как ценный гомозиготный материал. Вид Cucurbita pepo L. представлен огромным многообразием форм, включает высокопродуктивные овощные культуры и имеет широкое распространение в мире. Несмотря на большую экономическую значимость, создание эффективных протоколов, обеспечивающих стабильное получение удвоенных гаплоидов у этого вида остается актуальной задачей. Получение DH-растений представляет интерес не только по причине ускорения селекционного процесса, но и за счет реализации огромного потенциала гаметоклональной изменчивости, заложенной у этого высоко полиморфного вида. В обзоре рассмотрены основные использующиеся технологии получения удвоенных гаплоидов у овощных культур C. pepo: партеногенез in situ стимулированный обработанной/облученной пыльцой, гиногенез in vitro (культура неопыленных семяпочек in vitro) и андрогенез in vitro (культура пыльников/микроспор in vitro). Представлен анализ исследований, проведенных с начала открытия гаплоидных растений до современных достижений и оценки перспектив в области получения DH растений. Рассмотрены основные критические факторы, влияющие на эффективность каждой из технологий и ее отдельных этапов. Представлена разработанная технология получения удвоенных гаплоидов кабачка с использованием культуры неопыленных семяпочек in vitro, позволяющая получать до 55 эмбриоидов на 1 культивируемую завязь (28 эмбриоидов/100 культивируемых семяпочек). Для внедрения гаплоидных технологий в селекционный процесс необходимо полученные растения-регенеранты оценивать на уровень плоидности. Использование прямого подсчета хромосом в апикальных клетках может представлять определенную сложность у этого вида ввиду большого их количества (2n=40) и их малого размера. В зависимости от уровня оснащения лаборатории определение плоидности с использованием проточной цитометрии клеточных ядер и подсчета количества хлоропластов в замыкающих клетках устьиц в эпителии абаксиальной стороны листа, может быть более удобными методами. В обзоре рассмотрены перспективы использования молекулярных маркеров для оценки на гомозиготность при DH-технологиях, в том числе и у C. pepo.

Doubled haploids have been widely used worldwide in breeding programs and fundamental research as valuable homozygous material for about 100 years. The species Cucurbita pepo L. are represented by a huge variety of forms, include highly productive vegetable crops and have a wide distribution in the world. Despite the great economic importance, the creation of effective protocols to ensure stable production of doubled haploids in this species remains an urgent task. DH plants are of interest not only because of the acceleration of the breeding process, but also because of the realization of the huge potential of gametoclonal variability inherent in this highly polymorphic species. In this review, we analyzed the main technologies used for obtaining doubled haploids in vegetable crops of C. pepo: parthenogenesis in situ stimulated by treated/irradiated pollen, gynogenesis in vitro (unpollinated ovule culture in vitro) and androgenesis in vitro (anther/microspore culture in vitro). An analysis is presented of the research carried out from the beginning of the discovery of haploid plants to the current advances and evaluation of the prospects in the field of DH plant production. The main critical factors influencing the efficiency of each technology and its individual steps are considered. The developed technology of doubled haploids obtaining using non-pollinated ovary culture in vitro is presented. This technology allows to obtain up to 55 embryoids per one cultivated ovary (28 embryoids/ 100 cultivated ovules) To introduce haploid technologies into the breeding process it is necessary to evaluate the obtained plants for ploidy level. The use of direct counting of chromosomes in apical cells may present a certain difficulty in this species due to their large number (2n=40) and their small size. Depending on the level of laboratory equipment, ploidy determination using flow cytometry of cell nuclei and counting the number of chloroplasts in stomatal guard cells in the epidermis of the abaxial side of the leaf may be more convenient methods. The prospects for the use of molecular markers for assessment for homozygosity in DH technologies used, including C. pepo, are discussed in the review.

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