Получение удвоенных гаплоидов Cucurbita pepo L.

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.

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