Analysis of the preservation of the Quercus robur L. genotype using SSR- and ISSR-markers in the creation of callus cultures

Pedunculate oak (Quercus robur L.) is a forest-forming species in the European part of Russia and has a high economic value. However, at the same time, reproduction and cultivation of oak plantations causes significant difficulties. At present, a technique for clonal micropropagation of this species using nodal segments has been developed, but its application to old-growth trees causes difficulties. Therefore, it is relevant to search for alternative biotechnological approaches for the reproduction of adult oak material, including the production of regenerated plants through callus culture by indirect organogenesis or somatic embryogenesis. However, such a method of reproduction can lead to an intense manifestation of somaclonal variability, so its use should be accompanied by the control of the preservation of the genotype. This paper presents the result of the analysis of the preservation of the Q. robur genotype during the creation of callus cultures . Parts of green and semi-lignified shoots of a 200-year-old tree obtained during the growing season served as primary explants for obtaining oak callus cultures. To identify variability, two sets of molecular markers were used: microsatellite (SSR) and intermicrosatellite (ISSR). PCR was performed with the selected markers, and the amplification products were detected by agarose gel electrophoresis. As a result, the involved markers showed the complete identity of the genotypes of the original trees and callus tissues grown in in vitro culture. Thus, it has been shown that the cultivation of pedunculate oak calli has prospects for the conservation and reproduction of valuable tree genotypes.