OPTIMIZATION OF TECHNOLOGICAL SCHEME OF OBTAINMENT OF CULTIVATED CABBAGE (BRASSICA OLERACEA L.) DOUBLED HAPLOIDS UNDER CONDITIONS OF IN VITRO ANTHER CULTURE BY THE CONTROL OF GAMETE ORIGIN OF REGENERATED PLANTS BY PCR-ANALYSIS

S.V. Garkusha, N.V. Epifanovitch, Zh.M. Mukhina, E.G. Savenko, V.A. Glazyrina, L.A. Shundrina
All-Russian Rice Research Institute, p. Belozernyi, 3, Krasnodar, 350921, Russian Federation

Summary. The technology of guaranteed obtainment of doubled haploid lines of cultivated cabbage (Brassica oleracea L.) for their further use as parental forms is of great importance, as it allows accelerating several times the process of developing commercial F1 hybrids. The main purpose of this research was the development of laboratory testing methods for the control of gamete (from microspores) origin of cabbage regenerants, obtained in vitro culture, on the basis of molecular-genetic approach. The main principle of improvement, used for the first time, was a comparative genotyping of donor forms (source of explants) and regenerants, derived from explants, by molecular (microsatellite) co-dominant markers. It was obtained the regenerants of cultivated cabbage from various donor genotypes in anther culture in vitro and microsatellite profiles of donor genotypes by five SSR-markers. As a result, the Bol AB 20 TR SSR-marker was experimentally selected, which has co-dominant nature of inheritance in studied genotypes. PCR conditions with the selected marker were optimized for studied genotypes. Microsatellite profiles of regenerant plants, grown in vitro conditions from anthers of donor genotypes, in SSR-locus Bol AB 20 TR were obtained. The comparative characteristic of the allelic state of the microsatellite locus Bol AB 20 TR in donor and regenerant plants was carried out, and the conclusion about their gamete origin was made. The scheme is designed to ensure genetic purity and morphological evenness of F1 hybrids of cultivated cabbage developed by domestic breeders. After field testing, it will be in demand in practical breeding programs carried out, in particular, in the All-Russian Research Institute of Rice Breeding.

Keywords: cultivated cabbage, anther culture in vitro, gamete origin of regenerants, SSR-genotyping.

Author Details: S.V. Garkusha, D. Sc. (Agr.), director (e-mail: Этот адрес электронной почты защищён от спам-ботов. У вас должен быть включен JavaScript для просмотра.); N.V. Epifanovitch, post-graduate student; Zh.M. Mukhina, D. Sc. (Biol.), head of laboratory; E.G. Savenko, senior research fellow; V.A. Glazyrina, senior research fellow; L.A. Shundrina, senior research fellow.

For citation: Optimization of Technological Scheme of Obtainment of Cultivated Cabbage (Brassica oleracea L.) Doubled Haploids under Conditions of in Vitro Anther Culture by the Control of Gamete Origin of Regenerated Plants by PCR-Analysis / S.V. Garkusha, N.V. Epifanovitch, Zh.M. Mukhina, E.G. Savenko, V.A. Glazyrina, L.A. Shundrina. Dostizheniya nauki i tekhniki APK. 2016. V. 30. No 9. Pp. 33-35 (in Russ.).