Scientific Journal of KubSAU

In the process of the research we had accomplished an analysis of allelic polymorphism of self-incompatibility in pears in demand in the production and breeding of modern varieties of pears of North-Caucasian Zonal Research Institute of Horticulture and Viticulture collection of genetic resources. In the first stage the consensus primers PycomC1F and PycomC5R were used. With obtained data, after identification of the alleles studied in varieties using consensus primers an allele specific S5 and S8 DNA markers were used for confirming the presence / absence of data allele studied cultivars

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