Biomics

Medicago varia Mart. (bastard alfalfa) is a highly productive, protein-rich, perennial forage crop, well adapted to the conditions of the Southern Urals and Cis-Urals. In the Republic of Bashkortostan, the breeding of alfalfa is carried out at the Bashkir Scientific Research Institute of Agriculture of the UFRC RAS and the experimental station “Ufimskaya”. To assess the genetic polymorphism of the initial seed material, accurately identify and certify alfalfa varieties, the SSR analysis method can be used, which consists of PCR analysis of the sizes of loci containing microsatellite repeats. Most alfalfa SSR markers have been developed for the model object Medicago truncatula Gaertn. (truncated alfalfa), several studies on the SSR loci of Medicago sativa L (common alfalfa) have been published. For genetic analysis of bastard alfalfa, it is proposed to use the same SSR markers, but how effective they will be on domestic varieties of this species remains unknown. The purpose of this study was to test SSR primers for 10 microsatellite loci on populations of bastard alfalfa, which are in breeding study at the Republic of Bashkortostan. For the study, four samples of M. varia were used: variety Chishminskaya 131, populations Skorospelaya 9, S 3-6, and S 3-8. Analysis of the molecular genetic polymorphism of these M. varia samples using the SSR-PCR method allowed us to obtain data on the allelic composition of the microsatellite loci MsEST53, MSE265, MSE562, MSE486, MSE195, MSE590, MSE323, MsEST9, MSE470, MSE276. The most effective for genetic certification of varieties and populations of this crop were the SSR markers MsEST53, MSE265, MSE562, MSE486, MSE195, MSE590, MsEST9, which gave the largest number of polymorphic DNA fragments according to the results of PCR and polyacrylamide gel electrophoresis, making it possible to distinguish all four samples, and were also characterized by similar primer annealing temperatures and non-overlapping allelic ranges, which makes it theoretically possible to use them in multiplex PCR. The highly variable SSR markers identified during the study can be used to create test systems for accurate identification and certification of varieties, as well as for assessing the genetic polymorphism of M. varia samples.

Alfalfa, forage crop, DNA, PCR, SSR, microsatellite analysis

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