Optimization of marker system for genotyping oil flax varieties from the collection of VNIIMK

Abstract

Relevance. Currently, in Russia, oil flax (Linum usitatissimum L.) crop acreage is increasing, therefore, expansion of the range of varieties and rapid introduction of new, highly adaptive varieties into production are important tasks. Identification methods based on the use of genetic passports are useful both at the stage of studying parent material and when protecting breeders' copyrights. An optimal system for identification should cover evenly the entire genome. Therefore, the purpose of this work is the optimization of the existing marker system for genotyping oil flax varieties from the collection of All-Russian Research Institute of Oil Crops named after V.S. Pustovoit (VNIIMK) by increasing the number of the used polymorphic microsatellite loci. Materials and methods. 17 samples of oil flax from the VNIIMK collection were used as object of the research. Eight pairs of primers flanking the microsatellite loci were used as tools for the research. Localization of the studied primers in the reference genome of L. usitatissimum was determined using the web version of Primer-BLAST. DNA was extracted from two-week-old seedlings with CTAB buffer. The discriminative power of the marker system was determined using parameters such as polymorphic information content (PIC), frequency, the observed and effective number of alleles. Cluster analysis and graphical composition of dendrograms were carried out using Statistica 6.0 software package.Results. We determined the localization of the studied primers on seven chromosomes. Three pairs of primers were localized simultaneously on two chromosomes. Testing of the primers revealed 6 polymorphic loci. The number of observed alleles per locus ranged from 2 to 3 (average of 2.1 alleles per locus). The effective number of alleles ranged from 1.13 to 1.99 (average value —1.62), and the value of polymorphic information content (PIC) ranged from 0.111 to 0.498 (average value — 0.358). The extension of the set of polymorphic SSR loci allowed the differentiation of all used 17 genotypes.