E.V. Dubina, S.V. Koroleva, S.V. Garkusha
All-Russian Rice Research Institute, pos. Belozernyi, 3, Krasnodar, 350921, Russian Federation
Summary. The studies were carried out at the premises of the laboratory of biotechnology and molecular biology of the FGBNU All-Russian Research Institute of Rice Breeding (Krasnodar) in 2013-2016. The aim of the study was to develop a method for evaluation of genetic purity of seeds F1 of Capsicum annuum L., based on the polymorphism of microsatellite DNA markers. The material was sweet pepper genotypes used in breeding work of the department of vegetable and potato breeding of the FGBNU All-Russian Research Institute of Rice Breeding (Krasnodar) to create F1 hybrids Fisht and Pamir. Twelve codominant microsatellite (SSR) markers were approved, and the level of its polymorphic information content for 17 genotypes of sweet pepper was determined. To isolate DNA from plant samples we used achlorophyllous seven-day seedlings. DNA extraction was carried out by CTAB method. The oligonucleotide sequences of the codominant SSR-markers drawn from a database on www.VegMarks site. For the electrophoretic separation of PCR products it was used 8% polyacrylamide gel on the basis of 1x Tris-borate buffer. Visualization was performed in ultraviolet after staining the gels with ethidium bromide (EtBr). According to DNA analysis results it was selected three codominant microsatellite DNA loci (CAMS-117 – 11th chromosome, CAMS-142 – 1st chromosome and CAMS-405 – 8th chromosome) with the polymorphism level suitable for molecular genetics differentiation of the studied group of genotypes of Capsicum annuum L. (15-16 alleles). The level of hybridity of the hybrid F1 Pamir for loci CAMS-142 and CAMS-117 was 92% (it was 8 maternal plants per 100 studied plants), of Fisht – 96%. The determined high degree of polymorphism of the system of molecular marking based on DNA microsatellite loci enables to successfully use it as a reliable laboratory method for determining the level of hybridity of hybrids F1 of sweet peppers, as well as to assess the variety qualities of seed.
Keywords: sweet pepper, SSR-markers, molecular marking.
Author Details: E.V. Dubina, Cand. Sc. (Biol.), leading research fellow (e-mail: Этот адрес электронной почты защищён от спам-ботов. У вас должен быть включен JavaScript для просмотра.); S.V. Koroleva, Cand. Sc. (Agr.), head of division; S.V. Garkusha, Dr. Sc. (Agr.), director.
For citation: Dubina E.V., Koroleva S.V., Garkusha S.V. Use of SSR-Markers for Evaluating Hybrid Level of Sweet Pepper. Dostizheniya nauki i tekhniki APK. 2016. V. 30. No 8. Pp. 42-44 (in Russ.).