In this study, the genetic diversity of 64 Chinese Tetrastigma hemsleyanum Diels et Gilg germplasm accessions was analyzed by SRAP markers. By screening for polymorphism, ten out of 30 primer combinations generated reproducible fragments and they were used for additional diversity analysis in 64 genotypes. Fifty-seven polymorphic loci were amplified, with a polymorphic percentage of 66.67%-100% (mean, 90.38%). Each primer combination could generate 4-10, with a mean of 5.7. The average alleles (Na), the average effective allele number (Ne), the average Nei ''s gene diversity index (H) and the average Shannon diversity index (I) of 64 samples were 1.9038, 1.51501, 0.3009 and 0.45259, respectively. The genetic similarity coefficient ranged from 0.4603 to 0.9206. By UPGMA clustering, 64 samples were divided to 9 groups when the genetic similarity coefficient was 0.7256. Notably, several accessions collected from Jiangxi were clustered into group of accessions in Hubei, Hunan, Fujian and Guangxi, and several accessions could be clarified into group in Zhejiang. That indicated that Jiangxi may be the center or the transition zone of genetic diversity of Tetrastigma hemsleyanum Diels et Gilg germplasm resources. In addition, we constructed the fingerprints of 64 genotypes by using primers EM3ME3, EM13ME13 and EM14ME14. The dataset might provide reference in evaluation and identification of Tetrastigma hemsleyanum Diels et Gilg germplasm resources, and also breeding for new varieties .