POLYMORPHISM ASSESSMENT AND SELECTION OF SSR AND IPBS MARKERS FOR BETULA PENDULA MOLECULAR GENETIC STUDY
Abstract
Introduction. To use the molecular markers successfully, it is obligatory to select carefully the type of DNA marker and the particular primers used in the polymerase chain reaction. To solve the considered problem, it is necessary to conduct the preliminary research to detect the most informative DNA markers that allow determining the deepest polymorphism of the studied object. The goal of the research is to assess the level of variability of SSR and iPBS markers, and select the most informative primers for the genetic research of betula pendula. Six pairs of SSR primers and 14 iPBS primers were the object of the research. The methodology of the research included the following actions, implemented consistently: performance of polymerase chain reaction (PCR) with all the primers, visualization of PCR results using the electrophoresis, definition of the length of alleles of SSR loci in SSR analysis and PCR fragments using the iBPS primers, calculation of genetic polymorphism figures. Research results. The tested SSR and iPBS primers substantially differ in terms of the level of polymorphism. Number of the observed alleles for different SSR-locuses varied from 4 to 14; the observed heterozygosity was 0.38-0.88; the expected heterozygosity - 0.56- 0.89. The values of indicators of genetic polymorphism of 12 iPBS primers for Betula pendula were defined for the first time. The following figures for the indicators of variability (number of PCR fragments 7–17, number of polymorphic PCR fragments 3–17; share of polymorphic loci 42.9–100.0 %; number of observed alleles 1.43–2.00; expected heterozygosity 0.14–0.34) were typical for the iPBS. Conclusion. To conduct the genetic analysis, it is necessary to use, above all, the primers that allow revealing the deepest polymorphism of betula pendula. Using the studied SSR primers L1.1, L2.3, L5.4, and L7.8 the SSR loci were found to have had the largest number of alleles (8 -14). Among the tested iPBSprimers, iPBS2076, iPBS2078, iPBS2232, and iPBS2271 primers are of greatest interest, application of these primers made it possible to reveal from 10 to 17 polymorphic PCR fragments.
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