DNA markers are widely used in grapevine breeding to create forms with combined resistance genes. Downy mildew is one of the most common fungal diseases of the vine in the world. Growing grapevines with increased resistance allows to reduce the number of chemical treatments. The decrease in the use of pesticides is especially significant for viticulture of table varieties, since berries are directly consumed by humans for food. Currently, more than 20 resistance genes have been identified by molecular methods, and DNA markers for many genes have been developed. The genes Rpv3 (inherited from North American grape species) and Rpv12 (derived from V. amurensis) are among the most effective and have an additive effect. The study of 14 table grape varieties for the presence of the Rpv3 gene and 8 varieties for the presence of the Rpv12 gene was performed by using DNA-marker analysis. The analysis included varieties that could inherit these genes from the parent forms, according to their ancestry. The study was conducted using an automatic genetic analyzer ABI Prism 3130 and special software GeneMapper and PeakScanner, DNA-markers were taken from literature sources. According to the results of DNA-marker analysis, 9 varieties were identified, including 2 seedless varieties, with the Rpv3299-279 allele in the genotypes, which determines resistance to downy mildew, and 3 table varieties with the Rpv12 gene in the genotypes. One table grape genotype was identified with Rpv3 and Rpv12.
Local grape cultivars from different countries of the world are an important part of the gene pool of this culture. Of particular interest are the genotypes of the most ancient regions of viticulture. The territories of the subtropical zone of Georgia and the central part of Abkhazia belong to one of the centers of origin of the cultural grapevine. The purpose of the work was to genotype native Abkhazian grape cultivars, to study their genetic diversity based on DNA profiling data and to compare them with the genotypes of local varieties of other viticultural regions. Samples of plants were taken on the territory of the Republic of Abkhazia in private farmsteads and in the collection of the agricultural firm “Vina i Vody Abkhazii“ (“Wines and Waters of Abkhazia”). The genotyping of the Abkhazian cultivars Avasirhva, Agbizh, Azhapsh, Azhizhkvakva, Azhikvaca, Atvizh, Atyrkuazh, Achkykazh, Kachich was carried out using 14 DNA markers, 9 of which are standard microsatellite markers recommended for the identification of grape varieties. To improve our knowledge about the sizes of the identified alleles, we used the DNA of grape cultivars with a known allelic composition at the analyzed loci. Statistical analysis of the data showed that the observed heterozygosity for the analyzed loci exceeded expected values, which indicates a genetic polymorphism of the studied sample of varieties. Evaluation of genetic similarity within the analyzed group based on the results of genotyping at 14 loci showed that the cultivars Kachich and Azhapsh differed from the other Abkhazian varieties. The obtained DNA profiles of the Abkhazian cultivars were checked for compliance with DNA-fingerprints of grape varieties in the Vitis International Variety Catalogue. The Georgian varieties Azhizhkvakva and Tsitska turned out to be synonyms according to DNA profiles, two varieties from the Database (Italian Albana bianca and Georgian Ojaleshi) have differences in DNA-fingerprints from the varieties Atyrkuazh and Azhikvatsa only in one allele, respectively. When comparing the identified Abkhazian grape genotypes, their difference from the sample of Dagestan, Don, Greek, Turkish, Italian, Spanish, and French varieties and genetic similarity with the genotypes of Georgian grapes were shown.
Molecular genotyping of native varieties of Vitis vinifera L. from different winegrowing areas is a current trend in the grapevine genetic diversity research. Abkhazia is among the world cradles of tamed grape, and its indigenous gene pool is of particular interest. Avasirkhva is a native Abkhasian grapevine variety mainly grown in the Gudauta District. Te research aimed to obtaining a genetic passport of Avasirkhva grapevine using microsatellite polymorphism data. Te study sampled grape plants from private farmsteads of the Gudauta District. Te plant phenotype corresponded to the variety’s ampelographic description. DNA was isolated from young shoot tip leaves with a CTAB-based protocol. Genotyping was performed with polymerase chain reaction (PCR) followed by capillary fragment separation. High-polymorphic SSR loci (VVS2, VVMD5, VVMD7, VVMD25, VVMD27, VVMD28, VVMD32, VrZAG62, VrZAG79) recommended for grapevine varietal identification were used as markers. Te amplicon size was estimated with an ABI Prism 3130 automated genetic analyser using the GeneMapper and PeakScanner soſtware and Pinot Noir as a reference variety genotype. Four samples exhibited an identical microsatellite profile. Te microsatellite assay-based genetic passport of the Avasirkhva variety is as follows: VVS2141-145 VVMD5234-242 VVMD7239- 249, VVMD25239-249, VVMD27184-190, VVMD28234-248, VVMD32248- 262, VrZAG62200-204, VrZAG79251-257. Te obtained passport is unique with respect to the known genotypes in the Vitis International Variety Catalogue (VIVC). A Principal Coordinate Analysis of microsatellite data was used to infer the genetic relationships between Avasirkhva and the Abkhasian varieties Kachich and Azhizhkvakva genotyped in our earlier studies, as well as nine native grapevines of Georgia, the nearest viticultural area. Te Avasirkhva genetic passport can be used in grapevine genotyping studies to clarify the varietal identity.
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