ovoshchiОвощи РоссииVegetable crops of Russia2072-91462618-7132Федеральный научный центр овощеводства10.18619/2072-9146-2020-5-3-15ovoshchi-1133Research ArticleСЕЛЕКЦИЯ И СЕМЕНОВОДСТВО СЕЛЬСКОХОЗЯЙСТВЕННЫХ РАСТЕНИЙBREEDING AND SEED PRODUCTION OF AGRICULTURAL CROPSТехнология укоренения удвоенных гаплоидов редиса европейского, полученных в культуре микроспор in vitroRooting technique of double haploids obtained in culture of microspore in vitro for European radishКозарьЕ. В.KozarE. V.

Козарь Елена Викторовна – м.н.с. лаб. репродуктивной биотехнологии в селекции с.-х. растений

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

Elena V. Kozar – Junior Researcher of Laboratory of Reproductive Biotechnology in Crop Breeding

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

koz.leno4ek@gmail.comhttps://orcid.org/0000-0002-1319-5631КозарьЕ. Г.KozarE. G.

Козарь Елена Георгиевна – кандидат с.-х. наук, ведущий научный сотрудник лаб. иммунитета и защиты растений

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

Еlena G. Kozar – Ph.D. in Agriculture, leading researcher head of the laboratory of immunity and plant protection

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

https://orcid.org/0000-0002-9492-6845СолдатенкоА. В.SoldatenkoA. V.

Солдатенко Алексей Васильевич – доктор с.-х. наук, член-корр. РАН, гл.н.с.

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

Alexey V. Soldatenko – Dc. Sci. (Agriculture), corresponding member RAS, chief scientist

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

https://orcid.org/0000-0002-2695-190XДомблидесЕ. А.DomblidesE. A.

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

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

Elena A. Domblides – Ph.D. in 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 Federation20203010202005315Copyright © Козарь Е.В., Козарь Е.Г., Солдатенко А.В., Домблидес Е.А., 20202020Козарь Е.В., Козарь Е.Г., Солдатенко А.В., Домблидес Е.А.Kozar E.V., Kozar E.G., Soldatenko A.V., 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/1133Актуальность

Актуальность. Удвоенные гаплоиды (DH-растения), являются превосходным материалом для генетических исследований и селекции за счет полной гомозиготности. Род Rathanus в семействе Brassicaceae является самым не отзывчивым к технологии получения удвоенных гаплоидов с помощью культуры изолированных микроспор in vitro (IMC). Изучение причин нарушения корнеобразования у растений-регенерантов и отработка элементов этого этапа технологии, позволит значительно повысить эффективность IMC технологии для редиса европейского.

Методы

Методы. В исследование были включены три сортообразца из коллекции ФГБНУ «Федеральный научный центр овощеводства» (ФГБНУ ФНЦО): Тепличный Грибовский, Розово-красный с белым кончиком и Родос. В ходе экспериментов использовали стандартный протокол получения DH-растений с помощью IMC технологии в стандартном виде и с модификацией этапа укоренения. Для укоренения использовали твердую (агар 7 г/л) среду MS: безгормональная и MS с добавлением ИУК (в концентрациях 0,5; 1 и 2 мг/л) и жидкая питательная среда MSm с 0,1 мг/л кинетина. Концентрация сахарозы во всех средах составляла 20 г/л. Использовали три вида техники пересадки растительных эксплантов на агаризованную безгормональную среду MS: посадка микропобегов с погружением их базальной части на 2-3 мм в среду; посадка в лунку, сделанную в питательной среде с помощью пинцета в стерильных условиях; и посадка на поверхность среды без заглубления.

Результаты

Результаты. В настоящем исследовании изучены особенности этапа укоренения растенийрегенерантов редиса европейского в условиях in vitro. Показано, что для успешного укоренения микропобегов редиса важна техника пересадки. Растительные экспланты необходимо высаживать строго на поверхность твердой безгормональной питательной среды MS без заглубления. Использование для индукции корнеобразования жидкой среды MSm с добавлением 0,1 мг/л кинетина в пробирках с мостиками из фильтровальной бумаги также показала высокую эффективность. Для растений, склонных к образованию корнеплодоподобных структур (КС) с вторичными опухолями (ВО), необходима многократная диссекция аномальных образований с последовательными пересадками. Модификация этапа укоренения микропобегов повысила процент успешной адаптации DH-растений в условиях in vivo с 0-14% до 9598%.

Relevance

Relevance. Doubled haploids (DH-plants) are excellent material for genetic research and breeding due to their complete homozygosity. The genus Raphanus from the Brassicaceae family is the toughest to produce doubled haploid plants through isolated microspore culture in vitro (IMC). The study of the causes of disturbed root formation and the development of elements of this stage of technology will significantly increase the effectiveness of the IMC technology for European radish.

Methods

Methods. The study included three varieties from the collection of the Federal State Budgetary Scientific Institution Federal Scientific Vegetable Center (FSBSI FSVC): Teplichny Gribovsky, Rozovo-krasniy s belim konchikom and Rhodes. The experiments used a standard protocol for obtaining DH plants using IMC technology in a standard form and with a modification of the rooting stage. The solid MS medium (with agar 7g/L): MS without hormones, MS medium supplemented with IAA at concentrations of 0.5; 1 and 2 mg / L and liquid MSm medium supplemented with 0.1 mg / L kinetin were used for rooting of regenerated plants. All media were supplemented with 20 g/L sucrose. We used three types of techniques for transplanting plant explants onto a solid hormonefree MS medium: planting micro-shoots with their basal part immersed by 2-3 mm into the medium; planting in a well made in a nutrient medium using tweezers under sterile conditions; and landing on the surface of the medium without embedment.

Results

Results. In this work, we studied the features of the stage of rooting of regenerated European radish plants in vitro conditions. The transplant technique has been proven to be important for the successful establishment of radish micro-shoots. Plant explants must be planted strictly on the surface of a solid hormone-free nutrient medium MS, without embedment. The use of tubes with bridges made of filter paper and MSm liquid medium with the addition of 0.1 mg/L kinetin for the induction of root formation also showed high efficiency. For plants prone to the formation of root-like structures (RLS) with secondary tumors (ST), multiple dissection of abnormal formations with successive transplants is necessary. Modification at the rooting stage of micro-shoots growing has increased the percentage of successfully adapted DH plants in vivo conditions from 0-14% to 95-98%.

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