DNA analysis of centuries-old linden trees using SSR-markers

Abstract

Centuries-old trees are not only natural monuments, but also objects of biological, genetic and ecological research. The most numerous group of perennial trees in Ukraine, along with members of the genus Quercus L. is Tilia L. Genetic analysis of perennial linden trees will assess genetic polymorphism between different members of the same species, search for reference communities, and create genetic maps within a species. DNA markers have become widespread among molecular DNA labeling methods, in particular SSR markers. To assess the polymorphism of ancients Tilia cordata L. in research were used 6 SSR markers (Tc5, Tc915, Tc920, Tc927, Tc937, Tc963). The main stages and results of PCR technique for genetic analysis of perennial linden trees using SSR markers and determination of genetic polymorphism between them are highlighted. Our own universal SSR-PCR test system for DNA identification of genotypes of perennial lindens has been developed. According to the results of SSR genotyping, DNA profiles were obtained for 6 samples of historically valuable and centuries-old trees of Tilia cordata L. aged 400-1000 years. Based on microsatellite analysis, genetic relationships and the level of genetic diversity of selected genotypes were assessed. As a result of PCR, alleles of the expected size were obtained. According to the calculated value of the half-morphology index of the locus (RIC), the most polymorphic marker Tc920 PIC was 0.72, the lowest value of genetic distances (1.73). According to the results of studies of 6 samples of small-leaved linden by molecular SSR markers, molecular genetic polymorphism was determined in all studied samples. The analysis showed that the sample was characterized by a low level of genetic polymorphism, which may be due to the use of a limited number of parental forms in the selection process or genetically related forms.