RAPD diagnostics of barley varieties of domestic and foreign selection
Abstract
Purpose. To diagnose barley varieties of domestic and foreign selection based on protein content and isoenzymes by polymerase chain reaction using oligonucleotide RAPD primers, to establish the percentage of possible polymorphism of DNA fragments corresponding to target loci and to establish the relationship of varieties according to the studied traits. Methods. The research material consisted of barley varieties of domestic and foreign selection Acordine, Komandor, Bohun, Agrariy. DNA of the varieties was isolated from shade seedlings by the solid–phase method (DNA fixation on a magnetic sorbent). DNA amplification was carried out with a Bio-Rad C100 amplifier using primers UBC-402 and UBC-475 and subsequent detection in an agarose gel.Results. Assessment of the state of DNA preparations of barley varieties, carried out using gel electrophoresis, indicates the high quality of nucleic acid preparations, which is confirmed by thin bands (stripes). According to the results of polymerase chain reaction, we established the localization of genes associated with the expression of protein content traits and isoenzymes in the studied barley varieties. According to the first primer, the loci of the varieties Accordino and Agrariy have a length of 341 np, Komandor and Bohun – 258 np. According to the second primer, the loci are located in a slightly different way: Accordino, Komandor and Bohun – 258 np, Agrariy – 341 np. The results of our studies revealed that each target locus is complementary to one dekamer primer. Polymorphic fragments are absent at all. Another common feature of the studied barley varieties is the almost identical length of amplified DNA fragments.According to the results of cluster analysis, four separate clusters were formed, indicating the absence of kinship between the studied barley varieties according to the primers used. Conclusions. Our studies confirm the presence of polymorphic fragments of barley DNA according to two RAPD primers, but instead show the presence of loci of target genes responsible for the expression of protein content and isoenzymes. The sequence of fragments of a series of complementary primer complexes: GGGCGGCAAC and GGTCGCATAA. We also believe that it is necessary to conduct studies of the polymorphism of barley varieties using 10–20 RAPD primers, as well as ISSR primers.а
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