We present here the first large-scale genetic characterization of grape cultivars from Transcaucasia and Anatolia. These regions where wild grapes still grow in nature have been cultivating wine and table grapes for thousands of years and are considered the cradles of viticulture. Using 12 nuclear microsatellite markers, we genotyped 116 accessions of traditional grape cultivars from Armenia, Georgia and Turkey and we detected 17 identical genotypes and six homonymy cases, mainly within each national germplasm. Neighbour-joining analysis of genetic distance showed that each germplasm could have multiple origins and although they are now separated, they might have some common ancestors. In addition, four varieties from Western Europe included as outgroups turned out to be more related to Georgian cultivars than other germplasms, suggesting a possible ancient origin in Georgia. This work represents a first step towards germplasm management of this rich ampelographic heritage.