USING iPBS MARKERS TO ANALYSE WINTER BARLEY GENOTYPE POLYMORPHISM

The aim of the study was to evaluate the efficiency of using iPBS-molecular markers for genotype analysis of winter barley varieties of modern Russian selection. Fifteen winter barley varieties of different originators were selected for genotyping. Their genetic diversity was assessed using 10 iPBS-markers. To prepare samples for DNA extraction, winter barley seeds were pre-germinated in Petri dishes on moistened filter paper using a thermostat, excluding access to light. DNA extraction was performed using the CTAB-method, after which the extracted DNA was diluted with TE buffer and the concentration was checked using a Qubit fluorimeter. To ensure a high yield of the amplified product, the following polymerase chain reaction conditions were used: 5 min at 94 °C, then 40 cycles, 30 sec at 94 °C, 30 sec at 55 °C, 1 min at 72 °C, 3 min at 72 °C. Electrophoretic separation of PCR products was carried out for 1 h using 2 % TAE – agarose gel. The GelDoc gel-documenting system was used to visualize the separation results. As a result of the PCR and subsequent detection of its products in the range from 100 to 10,000 bp, 1223 alleles were clearly identified. Statistical processing of the obtained data made it possible to determine the degree of polymorphism of barley genotypes based on PCoA and cluster analysis, which in turn showed similar results, dividing the samples into four populations. The obtained results make it possible to use iPBS class marker systems to determine the degree of polymorphism of winter barley genotypes.