Anhui is one of the main areas of artificial crawfish (Procambarus clarkia) farming in China. To understand the genetic diversity of cultured P. clarkii populations in Anhui Province, this study selected three artificial cultured populations of P. clarkii in Anhui (WH, XC and HF) as the research object, with two wild population of P. clarkii in Anhui (TL and MAS) and four artificial cultured populations (JL, JH, GH and XH) as control. Ten pairs of P. clarkii microsatellite primers were selected to study microsatellite genetic diversity and genetic structure. The results revealed that the genetic diversity of P. clarkii artificial cultured in Anhui was higher than wild population and four artificial cultured populations (JL, JH, GH and XH), and the XC population had the highest genetic diversity. Tests of deviation from Hardy-Weinberg equilibrium indicated all 10 loci in 9 populations were significantly deviated from Hardy-Weinberg equilibrium after Bonferroni correction and most of them had significant heterozygosity deficiency．AMOVA analysis showed that genetic variation was determined within the population. F_st in most groups showed moderate differentiation (0.05 < F_st < 0.15). The gene flow demonstrated there were extensive gene exchanges between different populations, particularly between GH and JH. UPGMA tree revealed that XC and HF belonged to the same clade, and WU、GH、MAS and JH were in the other clade, whereas the remaining populations formed their own clade. STRUCTUR results showed that most individuals in XC and HF populations were assigned to the same genetic population, suggesting that they may have the same origin. The results showed that the cultured population of P. clarkii in Anhui had high genetic diversity, which provided reference for the protection and improvement of P. clarkii germplasm resources in Anhui.