Auricularia species are well known for their traditional edible and medicinal importance. In this study, sequence-related amplified polymorphism (SRAP) markers were used to evaluate the genetic diversity and relationships of 24 Auricularia strains obtained from different ecological regions of China. A total of 402 polymorphic loci were detected using 11 SRAP primer combinations with 100% polymorphic loci. The similarity coefficient ranged from 0.662 to 0.903, with an average of 0.73. Cluster analysis using the pair-group method using arithmetic average, based on the data from SRAP loci amplified by 11 primer pairs, showed that 24 Auricularia strains were distinguished into 4 groups with 73% similarity. The results of principal coordinate analysis were in accordance with pair-group method using arithmetic average clustering, and the first, most informative coordinate accounted for 74.38% of all the variations, which indicated a high level of genetic diversity among Auricularia strains. In general, SRAP markers provide a powerful tool to discriminate Auricularia strains and to evaluate their genetic relationships.