The aim of this study is to construct a DNA fingerprint database of 314 grape accessions, analyze the genetic diversity based on the genomic DNA simple sequence repeats (SSR), and to provide a scientific reference for the cultivar identification, genetic analysis and plant varieties protection. By using 9 SSR primer sets to analyze the 314 grape samples, 199 alleles were produced, of which the polymorphic loci were 199 and the ratio of polymorphism was 100%. Each SSR marker detected 17-31 alleles (with an average of 22.1). The polymorphism information content ranged from 0.793 to 0.886, with an average of 0.839. In this study, we suspected that 3 groups were homonyms and 9 groups were synonyms. In the remaining 290 grape accessions, seventy cultivars could be identified with one primer pair and the other cultivars could be identified with primer combinations. The 290 cultivars could be completely distinguished from each other with only 8 primer pairs. In summary, DNA fingerprint database of the 314 grape cultivars was constructed by using 8 SSR primer sets. Cluster analysis of the fingerprint database showed that the 263 diploids could be divided into two groups: Subgen. Euvitis Planch. and Subgen. Muscadinia Planch. The former group were further divided into 1 large group and 14 small groups. 51 polyploids were divided into 3 groups. The clustering results were basically in good agreement with the family tree of the tested cultivars.