Abstract: In this study，304 Vitis accessions were used as experimental materials for sequencing by specific-locus amplified fragment sequencing（SLAF-seq）. In total，466 618 high quality SLAF tags were obtained，including 392 374 polymorphic SLAF tags. A total of 481 192 single nucleotide polymorphisms （SNPs）were obtained by sequence analysis. The phylogenetic relationship and population structure of 304 Vitis accessions were analyzed based on the SNP dataset，which suggests that SLAF-seq can be used to develop a large number of SNPs for population genetic analysis，efficiently and cheaply. The clustering results showed that V. vinifera，Euro-America hybrid and wild species were distinguished from each other，among which sister lines and varieties with parent-child relationship were clustered together，respectively. In addition，the Chinese grape landraces have a close relationship with the ancient Eurasian grape varieties，and the ancient grape varieties with with unknown origin and genetic background are closer to the wild species than the improved varieties. These results provide references for further study of the origin andevolution of grapes，and provide empirical data for the establishment of SNP fingerprint database.

Key words: grape, SLAF-seq, SNP, genetic diversity