Genetic similarity of the autochthonous grapevine varieties from don region revealed by SSR-analysis and main leaf ampelographic traits

Native, ancient grape varieties of different cultivation regions are important part of grapevine genetic resources. Many native Don grape varieties represent a significant value for cultivation and use in breeding. The close varieties and more distant groups are distinguished on the main characteristics among the varieties of Don. The main features of the leaves of grape varieties are the key ampelographical characteristics. Currently, the study at the DNA level is considered the most informative method of plant genotyping analysis. Microsatellite markers are widely used for genotyping of grapevine varieties and rootstocks, and successfully applied in the study of the origin of varieties and the analysis of their pedigrees. We evaluated the relationship among the number of Don varieties by microsatellite genotyping. The aim was to study the genetic similarity of native Don varieties based on DNA analysis and compare the results with earlier made conclusions about relationship of varieties, and with data of the analysis of the main features of the leaves. The research was carried out on 16 varieties from the collection of the All-Russian Scientific Research Institute of Viticulture and Winemaking (Novocherkassk) and the Russian ampelographic collection (Anapa). Studied Don grapevine varieties were described ampelographically, and the main method we used in the work was the PCR. Six SSR-markers basicly recommended for V. vinifera fingerprinting were used. DNA was extracted from young leaves of the apical shoots of 4-5 typical bushes. Chardonnay and Cabernet Sauvignon were used as the reference cultivars. Genetic distance matrix was constructed using the coefficients (indices) similarity of M. Nei and W. Li. Based on the data of SSR-genotyping, estimation of the genetic similarity of studied varieties was performed using cluster analysis (UPGMA), and dendrograms were graphically constructed. Data on the morphological characteristics of leaves and SSR-genotyping results were analyzed by means of principal coordinates (PCA). DNA profiles of 16 local Don grapevine varieties were obtained using microsatellite loci VVMD5, VVMD7, VVMD27, VVS2, VrZAG62 and VrZAG79 with an automated genetic analyzer ABI Prism3130 («Applied Biosystems», USA). In the studied Don varieties genotypes, six (for VVS2, VVMD5, VMD7, VrZAG62) and seven (for VVMD27, VrZAG79) alleles per locus were determined. Cluster analysis allowed to divide the varieties into two main groups: one included Sibir’kovyi, Puhlyakovskii belyi, Sivolistnyi, Puhlyakovskii chernyi, Kosorotovskii and Kukanovskii cultivar, being a group of natural seedlings of Puhlyakovskii belyi, the other contained Bezymyannyi Donskoi, Plechistik oboepolyi, Staryi Goryun, Tsimlyanskii belyi, Tsimlyanskii chernyi, Tsimladar, Plechistik, Sypun chernyi, Mahrovatchik and Bessergenevskii № 7 cultivars. Interestingly, the second cluster had three subgroups. One includes varieties Bezymyannyi Donskoi, Plechistik oboepolyi, Tsimlyanskii belyi, Tsimlyanskii chernyi, Tsimladar, Plechistik, Sypun chernyi of the Tsimlyanskii group. The other contained Bessergenevskii № 7 cultivar being presumably a seedling of Puhlyakovskii belyi, and Stary Goryun of the Tsimlyanskii group. Variety Mahrovatchik (considered to be a seedling of Kokur white variety) was grouped separately. Analysis of the main features of leaves showed no differentiation according to the presumed origin of the studied varieties. As the result of SSR-analysis, most of varieties were distributed in accordance with the earlier made conclusions about their origin. Thus, the study of collections, old varieties, breeding material and introduced samples based on the complex of ampelographic traits and SSR-markers can be considered as the most informative one.