This study was conducted to investigate the genetic diversity and to develop a technique for cultivar identification using SSR markers in grapevine. Thirty Korean bred and introduced grapevine cultivars were evaluated by 28 SSR markers. A total of 143 alleles were produced ranging from 2 to 8 alleles with an average of 5.1 alleles per locus. Polymorphic information contents (PIC) were ranged from 0.666 (VVIp02) to 0.975 (VVIn33 and VVIn62) with an average of 0.882. UPGMA (unweighted pair-group method arithmetic average) clustering analysis based on genetic distances using 143 alleles classified 30 grapevine cultivars into 7 clusters by similarity index of 0.685. Similarity values among the tested grapevine cultivars ranged from 0.575 to 1.00, and the average similarity value was 0.661. The similarity index was the highest (1.00) between 'Jinok' and 'Campbell Early', and the lowest (0.575) between 'Alden' and 'Narsha'. The genetic relationships among the 30 studied grapevine cultivars were basically consistent with the known pedigree. The three SSR markers sets (VVIn61, VVIt60, and VVIu20) selected from 28 primers were differentiated all grapevine cultivars except for 'Jinok' and 'Campbell Early'. Five cultivars ('Narsha, 'Alden', 'Dutchess', 'Pione', and 'Muscat Hamburg') were identified by VVIn61 at the first step. Then 21 cultivars including 'Hongsodam' by VVIt60 at the second step and 2 cultivars ('Heukbosuck' and 'Suok') by VVIu20 at the third step were identified. These markers could be used as a reliable tool for the identification of Korean grapevine cultivars.