Fruit growing
and viticulture of South Russia

Annotation

The article presents genotyping results of 16 samples with indetermined status from the Ampelographic Collection �Magarach� by 9 nuclear microsatellite markers (ssrVVMD5, ssrVVMD7, ssrVVMD25, ssrVVMD27, ssrVVMD28, ssrVVMD32, ssrVrZAG62, ssrVrZAG79, ssrVVS2). Extraction of genomic DNA was carried out using a CTAB buffer from an average sample of young leaves or green cambial tissue of shoots. The purity and amount of DNA were assessed spectrophotometrically using a �Biophotometer plus� instrument. The polymerase chain reaction was performed on a �T100 Thermal Cycler� (BioRad). The PCR mixture consisted of a 2.5x reaction mixture and primers with fluorescent labels FAM, TAMRA, and R6G produced by �OOO Sintol�. The primers were combined into multiplexes in accordance with the size of the resulting amplicons and the annealing temperature. Fragment analysis of PCR products was performed on an ABI Prism 3130 genetic analyzer using Gene Mapper software. The varieties were identified by comparing the obtained DNA-profiles with the Vitis International Variety Catalog (VIVC) database. According to the results of genotyping, samples with indeterminate status are identified as varieties: Asyl Kara, Vasarga chernaya, Galbena de Odobeshti, Genusa Tsibil, Grec Rouge, Cruciulita, Marash Cherven, Muscat fleur d�oranger, Muscat Hamburg, Mtsvane Kakhuri, Negru Virtos, Ozirovka, Plavaie, Rkatsiteli, Saperavi, Tsolikouri. Deviations in the obtained DNA-profiles from the data of VIVC database by 1-2 alleles were revealed for 8 studied samples, which, presumably, can be explained by population variability, and in some cases by somatic mutations. This study demonstrates the result of molecular-genetic approaches to identify grape varieties.

Annotation

The paper presents the results of DNA profiling of 28 grape samples with unidentified varietal affiliation, selected from the Magarach Ampelographic Collection, and provides polymorphism assessment of their microsatellite loci. Genotyping was performed using 9 nuclear (VVMD5, VVMD7, VVMD25, VVMD27, VVMD28, VVMD32, VrZAG62, VrZAG79, VVS2) and 3 chloroplast (ccmp3, ccmp5, ccmp10) SSR markers. For nuclear loci, 105 alleles were identified, the average number of alleles per locus (na) was 11.7; the average effective number of alleles (ne) is 7.23; Shannon information index (I) for the studied batch of samples was 2.13. The highest allele frequency was observed in samples with VVS2133 and VVS2135 (p = 0.250), VVMD5238 (p = 0.232), VVMD7247 (p = 0.232), VVMD25239 (p = 0.250), VVMD27180 (p = 0.232), VVMD28236 (p = 0.143), VVMD32272 (p = 0.304), VrZAG62188 (p = 0.179), VrZAG79251 (p = 0.232) alleles. The average value of actual heterozygosity Heto was 0.8333, expected heterozygosity Het� was 0.8712. For chloroplast loci, 7 alleles were identified: ccmp3106, ccmp3107, ccmp5104, ccmp5105, ccmp10114, ccmp10115, ccmp10116. Also 4 chlorotypes were identified: A (1 sample), B (9), C (13), D (5). A dendrogram of genetic similarity was constructed using the UPGMA method, which showed the presence of three clusters containing, respectively, 11, 11 and 6 samples. There was a uniform distribution of chlorotypes B and D over three clusters, and chlorotype C � over two clusters. It is concluded that the results can be used to assess the genetic diversity of Magarach Ampelographic Collection.