An optimization of determination of genetic purity parameters of sunflower lines basing on DNA microsatellite loci

The wide introduction of hybrids into production is mainly limited by complicated seed growing foreseeing maintenance a high level of genetic purity of parental lines. So the work purpose was to optimize determination of parameters of sunflower lines genetic purity basing on DNA microsatellite loci. We used 12 sunflower lines from collection of the Armavir experimental station of the All-Russian research institute of oil crops (Krasnodar region). A genomic DNA was isolated from 100 separate 5-7-days growing etiolated sunflower seedlings of each sample using STAB-buffer. The 11 pairs of primers for PCR showed stable repeating polymorphism of fractions of amplified DNA. Seven pairs of primers were chosen as a result of determination of parameters of sunflower lines genetic purity using DNA loci and criteria of polymorphism level. To optimize the method two pairs of primers (ORS 5 and Har 432) were united into multiplex set. The last five loci (ORS 559, Har 432, ORS5, ORS 18, ORS 1144) were amplified separately. Creation of mixture containing united aliquots of DNA of the separate seedling in a vial allows decrease analyses quantity. In case of detection the ways with several fractions on phoregram after PCR, DNA samples of this atypical mixture are amplified apart to ascertain amount of off-type seeds.