Genetic divergence among four populations of Dendrolimus spectabilis，three populations of Dendrolimus tabulaeformis and two populations of Dendrolimus superans were investigated using Inter-simple Sequence Repeats Polymophism (ISSR) markers. DNA extracted from 119 individuals were amplified using 13 ISSR primers with 14 to 17 nucleotides. The agarose gel electrophoresis revealed 195 clear and stable bands with the molecular weight ranging from 250-2500 bps. Each primer yields about 3 to 10 bands per individual. Shannon′index showed higher genetic diversity within D. superans and the genetic diversity of D. tabulaeformis was close to D. spectabilis, both of which had a lower diversity level.The clustering followed the order: first individuals within a population, then different populations of the same species. The four populations of D. spectabilis fell into three branches, and two geographically close populations from Shenyang and Jinzhou were clustered into one branch ; the other branch was Tai′an population, and it was grouped to the third branch, Qingdao population. The two populations of D. tabulaeformis from Chengde and Pingquan counties were first clustered into a branch and then clustered with Shenyang population. The left two populations of D. superans species were clustered into one branch.