No abstract

Forage annual and perennial grasses are the difficult subject for molecular and genetic studies because of the problem with obtaining qualitative genomic DNA for PCR, due of high content of proteins, polysaccharides and polyphenols. The known methods of DNA extraction or the numerous commercial kits allow isolating purified nucleic acids from the leaf tissue, but characterized by low efficiency at seedlings using. The modified method of DNA isolation, based on the SDS-extraction buffer (sodium dodecil sulfate), is presented in this study. Significant modifications were introduced in the reagents compound and the steps of procedure accordingly to used type of plant tissue and the result was positive at usage on the bulking samples, as well as on the individual genotypes (the only seedling). Reliability of this method and the functionality of the obtained DNA samples were tested in PCR with different molecular markers (SSR, SRAP and PawS) in researches on revealing of forage legume grasses DNA polymorphism. The general advantages of the proposed method are simplicity and effectiveness, the possibility to isolate qualitative DNA without toxic reagents application, as well as relatively low cost and availability of reagents. This method can be useful for studying the genetic biodiversity and for decision the different tasks, required the rapid analysis of large plant populations.

The results of experimental studies on polymorphism of 10 red clover cultivars of Russian selection are presented. Informative primer pairs that can be used for the identification of markers during intervariety differentiation were determined. Based on SSR fingerprints, molecular genetic formulas of the studied cultivars were compiled. Samples of genomic DNA of each cultivar were obtained from pooled aliquots of the plant tissue, each containing 30 seedlings. Eleven pairs of SSR primers developed for structural analysis of the red clover genome were used in genotyping. Unique amplification fragments were found with SSR locus RCS1307 for Trifon and Topaz cultivars, as well as with SSR locus RCS3095 for Trio and Mars cultivars, which can serve as identification DNA markers The results of this study are of practical importance for the molecular genetic certification of breeding achievements, which is necessary to assess the quality and genetic uniformity of seeds and to control hybridization in the breeding process. forage crops, red clover, DNA fingerprinting, SSR loci, cultivar identification This study was funded from the federal budget within the framework of a state assignment (project no. 0442-2019-0001 АААА-А19-119122590053-0).

The conditions for creating alfalfa samples MN-2 (from the Selena variety) and P-67 (from the Lugovaya variety) with increased resistance to Fusarium pathogens using the method of gamete selection are described. The method is based on the selection of gametes in vivo under the influence of a selective factor (culture filtrate) introduced into the generative organs at the early stages of their development using vacuum infiltration. The method was developed in the biotechnology department of the V.N. V.R. Williams. The accessions are included in the breeding process and are currently the accessory varieties. In order to identify differences at the genetic level between the created varieties and the original forms, a comparative analysis was carried out using 2 types of molecular markers based on the PCR method. DNA polymorphism was detected using SRAP primers, which may indicate a change in the structure of genes associated with economic and biological traits. Research is ongoing. When growing under the conditions of a selection and greenhouse complex of acid-tolerant plants of meadow clover, created using cellular technologies in vitro, the infestation by powdery mildew of 120 acid-tolerant plants was studied. No signs of damage were found in 10% of plants. The largest number of affected plants (36.7%) had a score of 3. It was found that a high infestation with powdery mildew (scores 4 and 5) significantly reduces the number of inflorescences in the bush to 83.4 and 75.7%, respectively. Therefore, in further studies to create a population of acid-tolerant plants, the genotypes of meadow clover were used without signs and with a low score of powdery mildew damage.

Печатается по постановлению Ученого совета ФГБУН «Ордена Трудового Красного Знамени Никитский ботанический сад-Национальный научный центр РАН» № 11 от «29» сентября 2020 г.