Abstract: In this research, the TP-M13-SSR primers of Prunus tenella Batsch. were designed, and the primers with high polymorphic were screened out for further research. Two sample of leaves were selected from five populations respectively to extract the genome DNA, and the amplification production were analyzed by using the capillary electrophoresis with fluorescent-labeled primer. It was calculated that the number of different alleles(Na), the number of effective alleles(Ne), Shannon''s information index(I), Observed heterozygosity(Ho), Expected heterozygosity(He) and Fixation index(F) of 12 pairs of TP-M13-SSR Primer for Prunus tenella Batsch. ranged from 0.656-0.859, 0-0.281, 3.796-9.580, 4-8, 1.213-2.014, -0.164-1 0.605-0.843, respectively. In the above primers, all of the information data of primer PT-6 was the highest except the Fixation index(F) , and totally 12 pairs of TP-M13-SSR primers contained abundant polymorphisms were choosed These primers’ amplified fragments ranged from 100 to 300 bp, most of which were dinucleotide repeat.