Получение удвоенных гаплоидов Brassica purpuraria

Relevance. In recent years vegetable crop Brassica rapa ssp. chinensis var. purpuraria (synonyms: Brassica campestris L. var.purpurea Bailey; Brassica rapa L. ssp. chinensis var. purpurea) is gaining popularity as an object of genetic and molecular researches, and as an economically valuable vegetable plant due to the high content of biologically active compounds and distinctive economically valuable traits. Effective technology for development DH-plants to accelerate the breeding process for this culture has not been developed yet, so research in this area is relevant.Materials and methods. The study included two varieties from the collection of Vavilov AllRussian Research Institute of Plant Industry (VIR): No. 1301 (China) and No. 1357(Netherlands). Both protocols standard unmodified and with addition of silver nitrate (AgNO3) in the medium for embryogenesis induction were used in experiments for production of DH plants from isolated microspore in vitro. Direct chromosome counting in meristem cells and flow cytometry were used to determine the ploidy of regenerating plants.Results. As a result of the study embryogenesis in B. purpuraria culture can develop with the use of a standard protocol as well as with the addition of silver nitrate that showed a positive effect on the induction of embryogenesis. The yield of embryoids varied depending on the genotype of the individual plant within the variety accession. The highest yield of embryoids was 40 embryoids/petri dish. The main problem at the stage of regeneration is that about half of the regenerating plants occurred to be albinos and were not viable. We show a high degree of spontaneous chromosome doubling in regenerated plants (all analyzed plants were doubled haploids). In total 38 regenerated plants were obtained from accession No. 1301. It was shown that four DH-plants had self-incompatibility after self-pollination, but seed progeny from other plants was obtained. The created material was taken for genetics study and breeding work. Актуальность. Овощная культура Brassica rapa L. ssp. chinensis (L.) Hanelt var. purpuraria (L.H. Bailey) Hanelt (синонимы: Brassica campestris L. var. purpurea Bailey; Brassica rapa L. ssp. chinensis var. purpurea) в последние годы набирает популярность как объект генетико-молекулярных исследований и как овощное растение ввиду высокого содержания биологически активных веществ и хозяйственно ценных признаков. Эффективной технологии получения DH-растений для ускорения селекционного процесса у этой культуры еще не разработано, поэтому исследования в этой области актуальны.Материалы и методы. В исследование включены два сортообразца из коллекции ФГБНУ «Федеральный исследовательский центр Всероссийский институт генетических ресурсов растений имени Н.И. Вавилова» (ВИР): №1301(Китай) и №1357(Нидерланды). В ходе экспериментов использовали стандартный протокол получения DH-растений в культуре изолированных микроспор in vitro и протокол с добавлением нитрата серебра (AgNO3) в питательную среду для стимулирования индукции эмбриогенеза. Для определения плоидности растений-регенерантов использовали прямой подсчет хромосом в меристемных клетках и метод проточной цитометрии клеточных ядер.Результаты. В результате исследования установлено, что эмбриогенез в культуре B. purpuraria возможен как при использовании стандартного протокола, так и с добавлением нитрата серебра, который положительно влияет на индукцию эмбриогенеза. Выход эмбриоидов варьировал в зависимости от генотипа индивидуального растения в пределах сортообразца. Максимальный выход эмбриоидов составил 40 эмбриоидов на чашку Петри. Обозначена основная проблема, возникающая на этапе регенерации: около половины растений-регенерантов оказались альбиносами и были нежизнеспособны. Показана высокая степень спонтанного удвоения хромосом в растениях-регенерантах. Всего было получено 38 растений-регенератов из сортообразца №1301. Выявлено, что у некоторых DH-растений при инбридинге проявляется самонесовместимость (4 шт), в то время как у остальных растений семенное потомство было получено. Созданный материал передан для генетических исследований и селекционной работы.

Production of doubled haplois in Brassica purpuraria

Relevance. In recent years vegetable crop Brassica rapa ssp. chinensis var. purpuraria (synonyms: Brassica campestris L. var.purpurea Bailey; Brassica rapa L. ssp. chinensis var. purpurea) is gaining popularity as an object of genetic and molecular researches, and as an economically valuable vegetable plant due to the high content of biologically active compounds and distinctive economically valuable traits. Effective technology for development DH-plants to accelerate the breeding process for this culture has not been developed yet, so research in this area is relevant.Materials and methods. The study included two varieties from the collection of Vavilov AllRussian Research Institute of Plant Industry (VIR): No. 1301 (China) and No. 1357(Netherlands). Both protocols standard unmodified and with addition of silver nitrate (AgNO3) in the medium for embryogenesis induction were used in experiments for production of DH plants from isolated microspore in vitro. Direct chromosome counting in meristem cells and flow cytometry were used to determine the ploidy of regenerating plants.Results. As a result of the study embryogenesis in B. purpuraria culture can develop with the use of a standard protocol as well as with the addition of silver nitrate that showed a positive effect on the induction of embryogenesis. The yield of embryoids varied depending on the genotype of the individual plant within the variety accession. The highest yield of embryoids was 40 embryoids/petri dish. The main problem at the stage of regeneration is that about half of the regenerating plants occurred to be albinos and were not viable. We show a high degree of spontaneous chromosome doubling in regenerated plants (all analyzed plants were doubled haploids). In total 38 regenerated plants were obtained from accession No. 1301. It was shown that four DH-plants had self-incompatibility after self-pollination, but seed progeny from other plants was obtained. The created material was taken for genetics study and breeding work