ovoshchiОвощи РоссииVegetable crops of Russia2072-91462618-7132Федеральный научный центр овощеводства10.18619/2072-9146-2020-6-67-72ovoshchi-1188Research ArticleСЕЛЕКЦИЯ И СЕМЕНОВОДСТВО СЕЛЬСКОХОЗЯЙСТВЕННЫХ РАСТЕНИЙBREEDING AND SEED PRODUCTION OF AGRICULTURAL CROPSРазработка HybProb молекулярного маркера на аллель L4 для маркеропосредованной селекции перца сладкого Capsicum spp. на платформе Real-TimeDevelopment of the HybProb molecular marker for the L4 allele for marker-assisted selection of sweet pepper Capsicum spp. on the real-time PCR platformГавришС. Ф.GavrishS. F.

Сергей Федорович Гавриш – доктор сельскохозяйственных наук, профессор, заместитель директора по науке

127006, г. Москва, ул. Садовая-Каретная, д.8, стр.6, этаж 2, помещение II, комната 5,7

Sergey F. Gavrish – Doc. Sci. (Agriculture), professor

8, Sadovaya-Karetnaya str., bld. 6, 2 floor, II, room no 5, 7, Moscow, 127006

gavrish@gavrish.ruБудылинМ. В.BudylinM. V.

Михаил Вячеславович Будылин – кандидат биологических наук, заместитель директора по молекулярной диагностике и биотехнологии

127006, г. Москва, ул. Садовая-Каретная, д.8, стр.6, этаж 2, помещение II, комната 5,7

Mikhail V. Budylin – Cand. Sci. (Biology), deputy director for biotechnology and molecular diagnistics

8, Sadovaya-Karetnaya str., bld. 6, 2 floor, II, room no 5, 7, Moscow, 127006

bmw@gavrish.ruВербаВ. М.VerbaV. M.

Вадим Михайлович Верба – кандидат сельскохозяйственных наук, заместитель директора по селекции на Ближнем Востоке

127006, г. Москва, ул. Садовая-Каретная, д.8, стр.6, этаж 2, помещение II, комната 5,7

Vadim M. Verba – Cand. Sci. (Agriculture), deputy director for breeding in the Middle East countries

8, Sadovaya-Karetnaya str., bld. 6, 2 floor, II, room no 5, 7, Moscow, 127006

verbavm@mail.ruОбщество с ограниченной ответственностью «Научно-исследовательский институт селекции овощных культур»РоссияScientific research institute of vegetable breeding Ltd.Russian Federation202026122020066772Copyright © Гавриш С.Ф., Будылин М.В., Верба В.М., 20202020Гавриш С.Ф., Будылин М.В., Верба В.М.Gavrish S.F., Budylin M.V., Verba V.M.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/1188

Актуальность. Одним из условий создания современного высокотехнологичного гибрида сладкого перца является наличие в нём устойчивости к вирусу мягкой крапчатости (PMMoV), относящемуся к роду Tobamovirus. Данный вирус способен распространяться вместе с семенами, что может приводить к серьезному экономическому ущербу. Основным методом контроля распространения данного заболевания является возделывание устойчивых F1 гибридов перца. На сегодняшний день единственным универсальным источником устойчивости сладкого перца ко всем штаммам PMMoV является аллель гена L — L4. Однако до сих пор не существует надёжной маркерной системы, которая могла бы быть интегрирована в MAS. Методы. На основе данных из базы NCBI нами была сгенерирована молекулярная диагностическая система на основе гибридизационных зондов. Валидация была проведена на семи образцах рода Capsicum из мировых банков гермплазмы. После валидации для тестирования нового маркера использовали расщепляющиеся поколения F2 и F3 ряда селекционных образцов. Результаты. Используя экспериментальную маркерную систему Fret4 нами было протестировано 7 эталонных образцов гермплазмы перца с полным соответствием пиков расплава заявленным аллелям гена L (L0/L1/L3/L4). Далее при помощи маркерной системы был проведён МАS на 10 сортопопуляциях общим числом в 10 тысяч растений для выявления гена L4. В результате нам удалось создать высокоточную и экономически эффективную маркерную систему. Данная маркерная система, основанная на гибридизационных зондах легко интегрируется в МАS, позволяя быстро и надёжно определять аллельное состояние L4 при селекции сладкого перца на устойчивость к Тобамовирусам.

Relevance. One of the conditions for creating a modern high-tech hybrid of sweet pepper is the presence of resistance to the Pepper mild mottle virus (PMMoV). This virus belongs to the genus Tobamovirus. This virus can spread through seeds, causing enormous damage to the sweet pepper crop. The main method of controlling of virus is the cultivation of F1 hybrids of pepper with genetic resistance. To date, the L4 gene is able to confer resistance to all known strains of this virus. However, there is still no reliable marker system that could be integrated into the MAS. Methods. Based on data from the NCBI, we generated a molecular diagnostic system based on hybridization probes. Validation was performed on seven pepper accessions from the world's germplasm banks. After validation, the new marker was successfully tested on 2540 plants of F2 and F3 generation. Results. Using the experimental marker system Fret4, we tested 7 reference samples of pepper germplasm with full correspondence of the melt peaks to the declared alleles of the L gene (L0/L1/L3/L4). Then, using a marker system, MAS was carried out on 10 populations of an number of 10 thousand plants to identify the L4 allele. As a result, we were able to create a highly accurate and cost-effective marker system for the most important pepper resistance gene. This marker system based on hybridization probes is easily integrated into the MAS, allowing fast and reliable determination of the L4 allele state in sweet pepper breeding to Tobamovirus.

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