METHOD FOR DNA IDENTIFICATION AND GENETIC CERTIFICATION OF PERENNIAL AND ANNUAL RYEGRASS VARIETIES BASED ON SSR- AND SCoT-MARKING SYSTEMS Russian patent published in 2024 - IPC A01H1/00 C12Q1/68 

FIELD: molecular biology; plant genetics.

SUBSTANCE: described is a method for DNA identification and genetic certification of perennial and annual ryegrass varieties based on SSR- and SCoT-marking systems. Method involves the use of multilocus SCoT markers, which mark DNA associated with coding regions of the genome, to assess genetic polymorphism, detecting PCR products by preliminary agarose gel electrophoresis and drawing up molecular-genetic formulas based on the results of the detected allelic diversity. DNA extraction is carried out using a modified SDS method, in which 5 M ammonium acetate is used as a protein coagulant, DNA extraction is carried out without steps of additional purification and washing of solutions with ethanol, variety-specific DNA spectra are obtained from germinated seeds taken in an amount of at least thirty of each variety using two marking systems: 7 microsatellite SSR-loci and 9 SCoT-markers, detecting DNA fragments unique for the sample of the analyzed collection, the sizes of which are verified by high-resolution capillary electrophoresis and/or selective sequencing, molecular-genetic formula of the variety, as well as morphobiological and economic characteristics of the variety are added to the passport.

EFFECT: method enables reliable and reliable differentiation and identification of varieties included in the State Register of the Russian Federation, and can be used in selection, seed growing, in the system of variety testing and registration, in the work of control and testing laboratories, in solving legal issues on protection of intellectual property rights.

1 cl, 7 dwg, 4 tbl, 6 ex