Identification of sunflower lines and hybrids bred at the Armavirskaya experimental station of VNIIMK: selection of optimal DNA-loci and PCR conditions

Lines and hybrids of Armavirskaya experimental station of All-Russia Research Institute of Oil Crops are not identified under DNA loci. Allele polymorphism of one and the same loci can be different for genetic materials originated from the various breeding centers, so it is necessary to seek SSR-primers determining polymorphism of DNA fractions. Creation of a system of molecular markers on a base of DNA loci for a separate laboratory requires for optimization of temperature regimens of DNA amplification with different primers. Thus, the purposes of this work were optimization of PCR conditions, search of optimal microsatellite loci and determination of stable recurred polymorphism of DNA amplified fractions to create a system of molecular markers suitable for identification of sunflower lines and hybrids bred at Armavirskaya experimental station. Seven hybrids and 12 lines from sunflower collection of the Armavirskaya experimental station of VNIIMK were used for research. 18 pairs of SSR-primers were selected for polymerase chain reaction. Three primers did not hybridize with a matrix DNA in no one annealing temperature. An optimal temperature of hybridization with matrix DNA was selected experimentally for 15 pairs. It is showed that 11 pair of primers for PCR revealed stable recurred polymorphism of amplified DNA fractions, this allows to use them for molecular-genetic identification of sunflower inbreds and hybrids developed at the Armavirskaya experimental station of VNIIMK. Analysis of electrophoretic spectra of microsatellite loci showed some lines had identic set of microsatellite loci. Unicity of analyzed lines is 58.3%. Seven studied hybrids had different allele set of microsatellite loci in genotypes. The further screening of SSR-loci for more correct sunflower genotypes identification is supposed.