Abstract:A microsatellite enriched library of P. trituberculatu was constructed following the FIASCO method. Seventy-one pairs of primers were designed according to the sequences containing microsatellite motifs. Twenty-one loci were polymorphic after the evaluation of 71 loci based on genetic diversity analysis of a wild population comprising of 30 individuals for. A total of 188 alleles were obtained with an average of 8.9 alleles per locus. The number of alleles per locus amplified was significantly different, ranging from 3 to 13. The number of alleles obtained for the loci Pot8, Pot37, Pot48, Pot53, Pot54, Pot66 was 11,12,12,11,13 and 11, respectively, and Pot46 got only three alleles. Allele size distribution was in the range of 131 ~ 312bp, which is basically in line with the design theory of primer product length. Twenty-one microsatellite loci of expected heterozygosity were in the range of 0.659 ~ 0.889, and PIC values were higher than 0.5. The results indicated that they have a high heterozygosity, which can be used for P. trituberculatus population genetic structure, species breeding, and germ-line assessment of microsatellite DNA.