Fruit growing
and viticulture of South Russia

Annotation

Oomycete Plasmopara viticola causes one of the most harmful diseases of grapes � downy mildew. In the areas of the humid climate of the Black Sea coast of the North Caucasus, the pathogen causes particular damage. In the form of epiphytotic, Plasmopara viticola develops 6-7 times of 10 years and can cause`s the losses from 50 to 100 % of yeld, despite the presence of a large number of fungicides that can inhibit the harmfulness of this disease. The aim of the work was to test the microsatellite DNA markers of GOB, CES, ISA, and BER to study the diversity of P. viticola populations parasitizing in vineyards of the Krasnodar Territory. The material for the study were grape leaves of various varieties affected by mildew. The leaves was taken in May-July 2019 at various points in the Krasnodar Territory. P. viticola DNA was isolated from the diseased tissue of grape leaves using the �CytoSorb� developed by �Syntol� specifically for the diagnosis of phytopathogens. A total of 8 samples of P. viticola DNA were extracted. The study was carried out using the classical method of polymerase chain reaction with optimization of the number and duration of cycles, as well as the concentration of reagents. The size of the amplified fragments of the GOB, CES, ISA, and BER loci was estimated using an ABI Prism 3130 automated genetic analyzer and using fragment analysis. Data analysis was carried out in the program Gene Mapper 4.1. The greatest polymorphism was detected by the GOB marker (15 types of alleles in eight studied samples). The GOB, CES, ISA, and BER markers can be used to study P. viticola populations wide spreading in the vineyards of the Krasnodar Territory.

Annotation

One of the most important scientific problems in the genetics of cultivated plants is study and conservation of genetic diversity. Local native cultivars of different regions are the important part of world grapes gene pool. Traditional ampelographic descriptions are supplemented by molecular genetic data in the study of the gene pool of grapes at the present time. Genotyping of the 14 Dagestan autochthonous grapes cultivars has done in our research: Asyl Kara, Bayat Kapy, Gimra, Gulaby Daghestanskiy, Joonga, Dubut, Mola Huseyn Tsibil, Rish Baba, Sarah, Tavlinskiy Pozdniy, Hatal Baar, Hop Halat, Khotsa Tsibil, Shavrany. DNA profiles were obtained by microsatellite loci VVMD5, VVMD7, VVMD27, VVS2, VrZAG62 and VrZAG79. Above-noted SSR-markers are recommended as the main for Vitis vinifera genotyping. Molecular genetic analysis was performed on an automated genetic analyzer that provides data, corresponding to the modern world requirements for genotype identification. Using of multiplex PCR was tested in our work. Multiplex kits allow to optimize the time spending on analysis of DNA, as well as significantly reduce implementation costs. VVMD27 locus showed the highest polymorphism in the study of this group of Daghestan cultivars: 9 alleles per locus are identified; locus VrZAG62 showed the smallest polymorphism � 5 alleles / locus. The cultivars of grapes with uncommon alleles at the studied loci were determined in the analyzed varieties group. The information of number of microsatellite markers VVMD5, VVMD7, VVMD27, VVS2, VrZAG62 and VrZAG79 for fingerprinting of analyzed samples of grapes cultivars is shown.

Annotation

The central part of Abkhazia is recognized as one of the regions where the cultural grapevine originates. Famous varieties that have created the fame of local wines as well as the less-studied genotypes and wild-growing forms grow here. The study of the local gene pool of grapes at the molecular genetic level makes it possible to more fully assess the genetic diversity of varieties and forms, to identify closer and distant genotypes. As a part of the study of the Abkhazia grapes, we are carrying out DNA profiling of local cultivars. The purpose of this work is to study the genotype of the variety Azhshkuakua (Azhizhkvakva) variety. The Azhshkuakua plants growing in the collection of the agricultural company "Wines and Waters of Abkhazia", corresponding to the varietal description was used in the work. DNA was extracted from the apical part of young shoots of the cultivar`s plants by the method based on the use of CTAB. Genotyping was performed using SSR markers recommended for identification of grape varieties: VVS2, VVMD5, VVMD7, VVMD25, VVMD27, VVMD28, VVMD32, VrZAG62, VrZAG79. The amplified PCR reaction products were evaluated by capillary electrophoresis using an ABI Prism 3130 automatic genetic analyzer, followed by sizing using the GeneMapper and PeakScanner software, correcting the values taking into account the data of the control (reference) genotype with a known allelic composition (Pinot noir). Analysis of the obtained DNA profile of Azhizhkvakva in the international database of DNA-fingerprint of grape varieties revealed its correspondence to the profile of the Tsitska cultivar, which is considered an indigenous variety of Georgia. The genotypes of these cultivars can be considered synonymous, since they have almost identical morphological features. According to the literature and molecular genetic data, confirmed the high degree of genetic similarity of varieties, it can be assumed that these cultivars are clonal variations of the same genotype.

Annotation

One of the limiting factors of consistently high yields with good quality are various diseases of the crop. For example, diseases caused by phytoplasmas can have a serious negative impact the quantity of the crop and its quality, which can further affect the quality of wine products and profits. Phytoplasmas are one of the most dangerous phytopathogens. On grapes, they are represented by two species � Candidatus Phytoplasma vitis Flavescence dorée (causes Golden yellowing of grapes) and Candidatus Phytoplasma solani Bois noir. Many plant diseases that are thought to be caused by phytoplasmas were described before molecular genetic studies identified the various groups of phytoplasmas that cause these diseases. It is now possible to assess the relationship between the classification of phytoplasmas and specific plant diseases. Golden yellowing of leaves is a quarantine disease for the Russian Federation and the European Union and it causes a great damage to vineyards, so quick and accurate identification of this disease is very important. The main method for identifying phytoplasmas is real-time PCR (RT-PCR) with specific primer systems. The aim of our study was to compare the methods for extracting DNA from plant tissue affected by phytoplasma for further real-time PCR. This study showed that the samples isolated using the commercial kits "AgroDiagnostics" and "CytoSorb" show the similar results, while the sample isolated by the laboratory method based on the use of CTAB buffer showed higher and earlier peaks on the graph, which is important for detecting a small amount of pathogen in the test material and proves the greater effectiveness of this method of isolation.

Annotation

Downy mildew is a vine disease caused by the obligate heterothallic biotrophic endoparasite Plasmopara viticola. The area of distribution of this pathogen are vineyards all over the world, but the greatest losses are in the viticulture zones with a temperate continental and subtropical climate. In Krasnodar Territory, epiphytotic development of mildew occurs 6-7 times in 10 years. The first molecular genetic studies of the pathogen began at the end of the 20th century. The purpose of this work is to evaluate, based on DNA marker analysis, the polymorphism of the P. viticola population in two generations of the pathogen on grape plants growing in an isolated geographic point. The material for the study was the affected grape leaves taken from the vegetation plot of the FSBSI NCFSCHVW, from plants without chemical treatment. The material was taken in two time intervals � the end of July (the first generation of the pathogen � 4 samples) and the middle of August (the second generation � 6 samples). To study the diversity, highly polymorphic SSR-markers � GOB and PV144, were used. P. viticola DNA was isolated directly from infected leaves by the CTAB method. A total of 10 DNA samples of the pathogen were analyzed. The study was carried out by the classical method of polymerase chain reaction. The size of the target fragments of the PV144 and GOB loci was estimated using an ABI Prism 3130 automatic genetic analyzer by fragment analysis. The data obtained were analyzed using Gene Mapper 4.1 software. The highest degree of polymorphism was found for the GOB marker � 7 types of alleles and to a lesser extent PV144 � 4 types of alleles. It was found that the samples of the pathogen population collected during the first generation have significantly higher genetic polymorphism, in contrast to the samples collected during the second generation. Research in this direction continues.