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A technology has been developed for DNA identification and certification of varieties of meadow clover (Trifolium pratense L.), alfalfa (Medicago varia Mart.), Sowing (M. sativa L.) and hop (M. lupuli-na L.) based on molecular analysis with using SSR and SRAP markers. The recommendations contain a description of the sequence of experiments and protocols for DNA typing procedures. The presented methods were developed by the authors on the basis of their own experimental research and using the data available in the literature. A characteristic of informative primers for each marking system is given, a set of DNA identification markers is proposed, and unique molecular genetic formulas of varieties are drawn up as the basis for a reference genetic passport. Methodological recommendations were prepared with the aim of mastering the technology of DNA certification of forage grasses in practice. Designed for managers and specialists of research and control laboratories, can serve as a textbook for students and postgraduates in specialized specialties.

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.