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

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

The study was performed to genotype some commercial wine yeast strains using the assay of Interdelta genomic sequences. Experimental parameters of PCR to identify were optimized and optimal simplified method of DNA extraction from dried preparations of yeast cultures was define. Proven method showed a high level of resolution and can be used for the analysis of genetic diversity wine yeast in combination with SSR-markers

In the course of the work, 33 ISSR markers were evaluated for efficacy in the detection of genetic changes in regenerants of Galanthus woronowii Losinsk.. Ten markers were found suitable for genotyping according to the species under study. Five samples from the selected ten were analyzed for a sample of 20 plants of regenerants and a mother plant. The obtained data testify to genetic stability of plant material in the process of microclonal propagation

The study was aimed on approbation of 11 ISSR DNA-markers for their using in DNA fingerprinting of rare plant species of the Western Caucasus Lilium Caucasicum Miscz. Ex Grossh., Galánthus woronowii Kolak., Pancratium maritimum L. according to results DNA-markers set was selected for further using in the investigation of genetic diversity of the mentioned plant species

As the result of the work, the test of microsatellite DNA markers for the identification of genomic polymorphism between apple cultivars Florina, Golden Delicious and clonal forms has been done. With the high level of intervarietal polymorphism, studied DNA markers showed no allelic differences between varieties and their clonal forms

Results of testing of multiplex sets SSR-markers for genotyping of rice varieties are presented in the article. Two sets of SSR-markers were formed: 1: RM1+ RM11+ RM70+RM122; 2: RM164+RM167+RM168. The optimal combination of DNA markers in the multiplex sets and PCR conditions allowed obtaining accurate, easily interpretable results when performing fragment analysis on automated genetic analyzer ABIprism3130. Using multiplex sets, genotyping was performed for several varieties of rice: domestic breeding and one variety – IR36 from the breeding of IRRI (Manila, Philippines). For all the studied varieties specific SSR-fingerprints were obtained. RM 168 marker showed in domestic varieties a low level of polymorphism - one allele of 97 bp. However, at the same time, the variety IR-36, showed a second type of allele 107 bp. In addition, the loci of RM1, RM11, RM167 and RM164 have unique alleles in this variety. It is consistent with significant genetic differences of these varieties and the rest of the varieties in studied sample. The proposed SSR multiplexes are promising for use in DNA certification of rice varieties and assessment of genetic diversity

Microsatellite DNA markers are currently used effectively in the study of the genetic diversity of the gene pool of fruit crops and DNA certification of varieties. For plum now there is rather limited list of works on the development of this type of DNA markers. Most often, the search for new SSR-markers for this species is carried out by checking of crossreproducibility of SSR-markers developed in other species of the genus Prunus. In this study, for the 18 SSR-markers previously developed on a peach, there was performed testing and evaluation of the prospects for the use of the genotyping of plum cultivars. Testing was made on the 4 varieties of genetically distant, belonging to the 4 different subspecies of Prunus domestica L., showed the effectiveness of their use. During the study, all tested DNA-markers were grouped together in multiplex sets comprising 3-4 markers. This allows simultaneous genotyping of 3-4 loci in a single PCR reaction. These multiplex kits are available for use in the study of genetic polymorphism of species Prunus domestica L

The study was performed to genotype some commercial wine yeast strains with SSR-markers. Five polymorphic SSR-markers were tested in a selection of 15 yeast strains. Tested SSR-markers showed a high level of informativeness as well as polymorphism and can be used further to analyze the genetic diversity of wine yeast

This study was aimed on testing of IRAP markers developed at plum on varieties of peach, Russian plum and cherry plum and genotyping of home and foreign varieties of plum, followed by an analysis of the data. During the testing of the markers have been identified as a high level of polymorphism between genotypes of plum and between the studied species. On the basis of the results obtained by genotyping 15 samples of Prunus was built dendrogram. Cluster analysis was divided into 3 groups corresponding varieties of P. persica, P. domestica and a common group for varieties of Russian plum and cherry plum