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 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

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

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

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

Professional apple gardening is bound to particular risks, of which is essential losses of a harvest because of diseases. An apple scab, the caused Venturia inaequalis (Cooke) G. Winter, brings the greatest loss. The main approach in monitoring of a scab of an apple-tree is creation of grades, steady against pathogen. In the present work we have carried out phytopathology testing of generations of the seedlings received from the free pollination of six forms of a crab of Malus orientalis from a collection MOS VIR (Maykop) characterized by a relative resistance to the apple scab in 2-3 classes of damage by long-term data. For infection we used inoculum, consisting both of natural population of a scab, and of strains of pathogen of various cultivars and geographical origin. When carrying out padding infection increase in force of an infectious background that can be bound to selection of the plants of biotypes of a fungus that are most adapted for genotypes was noted. It is recommended for precise elimination of unstable plants at selection at early stages of an ontogenesis to carry out more than one serial infection during the season. By results of the phytopathological testing, we highlighted three Malus orientalis forms from the six studied No. 17982, 17985 and 3080 the most perspective stability genes for an introgressiya in a cultural gene pool of an apple tree

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.

Industrial horticulture assumes the most effective use of the potential of varieties. One of the key factors determining the yield of garden plantings is the effectiveness of pollination. To obtain the maximum yield, it is necessary to ensure maximum pollination during the flowering period. For this reason, much attention has been paid to the selection of pollinators. Crab-apple forms are promising for use as pollinators, so this work was aimed at identifying the most common alleles of the self-incompatibility gene in the crab-forms using the molecular genetic method of analysis. The object of the study was 29 apple-tree creams and 3 elite selection forms. They carried out the molecular genetic identification of alleles S2 and S10, which are among the most common apple trees in the world gene pool. Allele S2 was identified in 16 samples (14 forms and 2 elite selection forms), while S10 allele in one sample (elite form 12/2-20 (24-28)). Data on the allelic composition of the S gene in the samples studied are of value for the formation of a genetic passport on the compatibility of the studied samples of apple with modern industrial varieties

Estimation of self-incompatibility allelic diversity in Russian apple varieties was conducted. We have established S-allele gene set for a number of varieties. The most common allele of gene was identified. Alleles frequency data obtained are consistent with those in the world apple germplasm

This article presents the results of testing IRAP DNA markers Cass1 and Cass2 applied to Prunus spinosa. The findings suggest the high perspectiveness of their using for the study of genetic diversity of the gene pool of this species. According to the results of the analysis of the sample 12 genotypes were identified from 6 to 13 fragments in the spectrum of Cass1 and from 5 to 11 fragments for Cass2. As a result of cluster analysis in the sample formed three groups of samples. In one of the groups, which is most distant from the other two, includes samples taken in Ukraine, while the remaining two groups included samples from Armenia, the Krasnodar region region, the Republic of Adygea, Ukraine and Moldova, and three cultural large-fruited form. The distribution of samples in clusters corresponded to their geographical origin that favors the objective assessment of genetic distances between the samples using Cass1 and Cass2 markers. Thus, it was concluded that the prospects of using DNA markers to study the genetic diversity within a species of Prunus spinosa