Abstract: Genetic diversity of 27 Pleurotus eryngii cultivating strains from different areas in China and Japan were analyzed to provide a molecular evidence for the identification and seed selection of Pleurotus eryngii. The tested isolates were studied using clustering analysis, combined with SRAP and ITS markers. 9 pairs of primers on 27 strains were selected, and 151 fragments were amplified, polymorphic bands were 130, the average polymorphic rate was 83.3%. The PIC (polymorphism information content) value of these markers varied from 0.27 to 0.42, averaging 0.36. Kinship of the tested strains was determined by analyzing their ITS sequences, and the result which was consistent with SRAP analysis, showed that part of the strains from the nearby areas were clustered with each other, their kinship was close, while some strains with large gaps in the geographical distribution were also clustered in the same phylogenetic tree branch, their genetic relationship may also be relatively close. Both of the two markers showed that the genetic diversity of the tested strains was relatively abundant, and there was a certain link between genetic similarity and geographical distribution. The method which combined SRAP with ITS could get better results, and it provided a powerful reference for the study on the genetic diversity of Pleurotus eryngii.