Abstract: Chengdu gray goat is an excellent local goat breed, which plays an important role in the improvement of goat breeds and the cultivation of new breeds in China. However, due to the lack of scientific breeding methods, some captive populations of Chengdu gray goat lead to the confusion of pedigree and inbreeding, which, to some extent, will lead to the degradation of the captive population. In the long run, the genetic diversity of Chengdu gray goat will gradually lose and some unique important genetic resources will gradually disappear. In this study, the genetic diversity assessment of Chengdu gray goat was conducted using the microsatellite markers. The results were as follows: (1) Eight pairs of microsatellite markers with medium and high polymorphism were successfully developed by cross-species method; (2) A total of 60 alleles were detected, with an average allele of 7.5, an average effective allele number of 2.36, an average observed heterozygosity of 0.577, an average expected heterozygosity of 0.634, and an average polymorphism information content of 0.580. The results showed that the genetic diversity of Chengdu gray goat population was high and it has high conservation potential.