DNA-fi ngerprinting of local Аbkhazian grape variety Kachich

Abstract

The study of the indigenous gene pool of Vitis vinifera L. of diff erent zones of viticulture with the involvement of molecular genetic methods is an urgent due to the wide diversity of grape varieties. The territory of Abkhazia is a unique zone, one of the hotbeds of the emergence of cultural grapes. «Kachich» is the oldest Abkhazian grape variety, known for the quality of wines obtained from its harvest. The variety is more resistant to fungal diseases compared to popular European varieties cultivated in modern Abkhazia, and is particularly resistant to rot of berries. However, often the local population calls old bushes by the name «Kachich», which give grapes of good quality for making of red wines and not always these forms correspond to the morphological description of the variety. The aim of the research is DNA fi ngerprinting of the «Kachich» grape variety based on the analysis of polymorphism of microsatellite loci. The object of the study was the plants corresponding to the variety description, growing in the collection of the agricultural company “Wines and waters of Abkhazia” (Sukhum, Abkhazia). DNA was isolated from the young leaves of the top of the shoot of typical plants of the variety by the CTAB-method. Genotyping was carried out by polymerase chain reaction (PCR) with separation of reaction products by capillary electrophoresis. Highly polymorphic SSR-markers recommended for grapes varieties identifi cation were used for study (VVS2, VVMD5, VVMD7, VVMD25, VVMD27, VVMD28, VVMD32, VrZAG62, VrZAG79). The size of amplifi ed fragments was estimated using the automatic genetic analyzer ABI Prism 3130 and special software GeneMapper and PeakScanner. To clarify the size of amplifi ed fragments, the DNA of reference varieties was used. According to the results of microsatellite analysis, the DNA-certifi cate of the genotype of the «Kachich» grape variety was compiled: VVS2153 155 VVMD5234 240 VVMD7239 249 VVMD25239 267 VVMD27186 193 VVMD28234 248 VVMD32262 272 VrZAG62194 196 VrZAG79236 236. The resulting DNA-profi le was checked for matches in the Vitis International Variety Catalogue VIVC. No matches of the obtained DNA-profi le with any other DNA-profi le presented in the Database were found. DNA-certifi cate of genotype «Kachich» can be successfully used for varieties identifi cation of grape plants if necessary to clarify their varietals affi liation.