vavilovВавиловский журнал генетики и селекцииVavilov Journal of Genetics and Breeding2500-3259Institute of Cytology and Genetics of Siberian Branch of the RAS10.18699/VJ15.069vavilov-445Research ArticleСТРУКТУРНАЯ КОМПЬЮТЕРНАЯ БИОЛОГИЯ И ФАРМАКОЛОГИЯCytoplasmic heredityОпределение типа цитоплазмы у растений семейства Капустные (Brassicaceae Burnett) с помощью ДНК маркеровIdentification of cytoplasm types in accessions of the family Brassicaceae (Brassicaceae Burnett) with DNA markersДомблидесЕ. А.DomblidesE. A.edomblides@mail.ruДомблидесА. С.DomblidesA. S.ЗаячковскаяТ. В.ZayachkovskayaT. V.БондареваЛ. Л.BondarevaL. L.Федеральное государственное бюджетное научное учреждение Всероссийский научно-исследовательский институт селекции и семеноводства овощных культур, Московская область, Одинцовский р-н, пос. ВНИИССОК, РоссияРоссияAll-Russian Research Institute of Vegetable Breeding and Seed Production, Moscow oblast, Odintsovo region, VNIISSOK, RussiaRussian Federation201502122015195529537Copyright © Домблидес Е.А., Домблидес А.С., Заячковская Т.В., Бондарева Л.Л., 20152015Домблидес Е.А., Домблидес А.С., Заячковская Т.В., Бондарева Л.Л.Domblides E.A., Domblides A.S., Zayachkovskaya T.V., Bondareva L.L.This work is licensed under a Creative Commons Attribution 4.0 License.https://vavilov.elpub.ru/jour/article/view/445

Идентификация типа цитоплазмы с использованием молекулярных маркеров позволяет упростить отбор пар для скрещивания при создании гибридов на основе цитоплазматической мужской стерильности (ЦМС). На 40 образцах семейства Капустные (Brassicaceae Burnett) для определения типа цитоплазмы были протестированы 22 пары уже известных праймеров, отобранных из литературных источников. С использованием стандартной и мультиплексной ПЦР были идентифицированы 7 различных типов цитоплазмы (Ogura, Ogu-NWSUAF, nap, pol, cam, rad, ole) у коллекционных образцов. Расшифровка нуклеотидной после- довательности продуктов, дифференцирующих различные типы стерильности цитоплазмы, подтвердила наличие в митохондриальной ДНК (мтДНК) исследуемых стерильных образцов локусов генов orf138, orf222, orf224, связанных c проявлением признака ЦМС. У стерильного образца капусты пекинской в мтДНК присутствовало сразу два гена, orf138 и orf222, что соответствовало типу Ogu-NWSUAF. У всех образцов, несущих цитоплазму Ogura, в мтДНК присутствовал участок 417 п.н., на 100 % гомологичный с митохондриальным геном orf138 – типа А (всего ранее выделяли 9 типов гена orf138 – от A до I). Исключение составил образец капусты белокочанной F1 Tekila, у которого был выявлен новый аллельный вариант гена orf138, характеризующийся наличием делеции в 39 п.н., но в отличие от orf138 – типа F, для которого также характерна делеция в 39 п.н., у него присутствовали две несинонимичные замены: A→T по 95-й позиции и G→A по 99-й позиции.

Identification of cytoplasm types with the use of molecular markers enables to simplify the parent line pair selection for hybrid development based on CMS. Forty accessions of the family Brassicaceae (Brassicaceae Burnett) were analyzed with 22 pairs of PCR primers taken from previous research. Seven types of cytoplasm (Ogura, Ogu-NWSUAF, nap, pol, cam, rad, ole) were observed among breeding accessions using standard and multiplex PCR technique. The sequence of PCR products that differentiated different types of sterile cytoplasm confirmed the presence of three genes, orf138, orf222, and orf224, in mitochondrial DNA associated with exhibition of CMS. The accession of Nappa cabbage (Brassica rapa Pekinensis Group) had two genes, orf138 and orf222, which corresponded to cytoplasm Ogu-NWSUAF. All accessions carrying Ogura cytoplasm had the 417 bp fragment, which was 100 % identical to the mitochondrial orf138 gene sequence corres- pon­ding to Type А (formerly nine sequence Types, from A to I, had been identified). An exception was the accession of white head cabbage Tekila F1 with a new allelic variant of orf138 gene, which combined the 39-bp deletion typical of the Type F orf138 sequence and two nonsynonymous substitutions, A→T and G→A, at positions 95 and 99, respectively.

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