Поиск и анализ нуклеотидных последовательностей в геноме подсолнечника, ассоциированных с генами устойчивости к расам заразихи Orobanche cumana Wallr.

Abstract

Studying of sunflower lines resistant to broomrape race G, bred in V.S. Pustovoit All-Russian Research Institute of Oil Crops (VNIIMK), Krasnodar showed the genetic control of resistance is monogenic with incomplete dominance. The purpose of the research was to seek and analyze molecular markers for the genes of resistance to broomrape race G using PCRmethods. In our research we used six developed in VNIIMK lines-donors resistant to broomrape race G: RGP1, RGP2, RGB, RGL1, RGL2, RGA, and susceptible lines VK 101, VK 678, VK 680. We extracted DNA from the leaves of young sunflower plants using STAB-buffer. For PCR-analysis we used 17 pairs of primers of two types: SCAR (sequence characterized amplified region) and SSR (simple sequence repeat). Conditions of amplification are as recommended by authors of markers with experimental selection of a temperature for primers hybridization. Within the research we balanced involved into work SSR- and 12 SCAR-loci using BLAST and a set of HanXRQr1.0. Due to these data we composed a physical map of loci location. After assessment of six resistant and three susceptible parental lines, seven markers demonstrated polymorphism by a length of locus DNA and two ones – by presence/absence of amplified fragment. For markers ORS 683 and ORS 1112 we observed a relation between loci polymorphism and susceptibility of parental lines to broomrape. This was certified with a presence of an allele of 364 n.p. length in ORS 683 and an absence of an allele of 375 n.p. length in ORS 1112. These markers were noted as primary ones for hybridological analysis in F1 и F2 with selected pairs for crossing. Additional markers will be selected individually using obtained data on loci polymorphism. Thus, as a result of the research we created a physical map for the further markers selection, found hybrid combinations and markers from the studied ones for hybridological analysis.