Efficiency of SSR and PawS markers for evaluation of genetic polymorphism among red clover (Trifolium pratense L.) cultivars

Background. Identification of crop varieties is presently one of the most important aspects due a significant annual increase in the number of newly developed cultivars. Application of molecular markers makes it possible to identify cultivars and secure protection of plant breeders’ rights. Marker techniques based on SSR loci and PawS markers were evaluated for their efficiency in revealing the DNA polymorphism in Russian red clover cultivars, and the research results are presented in this publication.

Materials and Methods. The total genome DNA was extracted by a modified SDS method from 30 seedlings per each cultivar. Nine simple sequence repeats (SSR) and 4 PawS markers were used for genotyping. The basic genetic diversity parameters were measured and analyzed using the software resources GelAnalyzer 2010а, MStools v.3, and Statistica 7.0.

Results and conclusion. The mean level of intervarietal DNA polymorphism in red clover was 38.6%. Cultivar-specific amplicons were obtained for 4 accessions (cvs. ‘Trifon’, ‘Topaz’, ‘Trio’ and ‘Mars’) with SSR loci RCS1307 and RCS3095. These loci were found appropriate for identification and certification of such cultivars. The tested PawS markers (individually and in combinations) proved non-informative for the analysis of intervarietal DNA polymorphism in red clover. The only primer pair PawS5+PawS16 generated reproducible PCR products, but unique amplicons were absent in the DNA profiles. The data obtained in this study may be helpful for further identification and certification of Russian red clover cultivars and promising breeding materials. 

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