Scientific Journal of KubSAU

Isolation of high-quality RNA from the tissues of perennial woody plants, including woody grape vines, is very difficult due to the high content of phenolic compounds, secondary metabolites and polysaccharides and the ribonuclease activity of destroyed tissues. Most of the existing methods require either large time or financial costs, or do not give reproducible results in the case of RNA extraction from mature grape tissues. The modified isolation protocol is based on a combination and modification of the known RNA extraction methods, taking into account the characteristics of mature grape tissues. Existing commercial kits for the isolation of RNA from plant tissues showed a low efficiency of RNA extraction from mature grape tissues, primarily associated with "varietal specificity". Reproducible results in the extraction of RNA showed CTAB-method, however, it has several significant drawbacks associated with the duration of the extraction and the complexity of the processing of an RNA preparation with a DNAase. The developed method is based on increasing the concentration of mercaptoethanol and polyvinylpyrrolidone in the extraction buffer, eliminating the stage of RNA selective precipitation via LiCl, and replacing it with deposition on a silica-based membrane (SiO2) followed by processing with DNA-ase. and increase the purity of the preparation of RNA from genomic DNA in comparison with the original method. A modified isolation protocol was developed based on a combination and modification of known RNA extraction methods, taking into account the characteristics of mature grape tissues. This solution allows to obtain reproducible quantity and quality of RNA for the subsequent synthesis of cDNA and RT-PCR

Genetic studies of apricot are the actual direction in the genetics of fruit crops. In this regard, the improvement of the collection of SSR markers for the genotyping of this culture is an objectively significant task. In a study for the 16 SSR-markers previously developed on almonds (PdUnchar2, PdSLD1, PdGMGT1, PdTrTFGT1, PdUnchar2, PdSLD1, PdGMGT1, PdTrTFGT1) and Siberian apricot (A3-72, A1-63, H2-22, A3- 7-1, H2-5, A1-7, A3-9, H2-45), approbation and evaluation of the prospects of using for genotyping Prunus armeniaca L. were performed. Approbation, performed on 3 varieties of different origin, revealed markers and their combinations optimal for their use. During the study, all tested DNA markers were grouped into multiplex sets, including 4 markers. This allows carrying out genotyping simultaneously on 4 loci in the formulation of one reaction. One marker (PdUnchar2) from the studied sample included in the multiplex set did not show amplification. Five markers gave a monomorphic product. The remaining 11 SSR markers allowed us to obtain polymorphic, cultivar-specific SSR fingerprints for all the studied cultivar. These multiplex sets are proposed for use in studying the genetic polymorphism of the species Prunus armeniaca L.