Abstract: Four hybrid strains of Hypsizigus marmoreus obtained in previous experiments were used as the main test materials.The superior strain of Hypsizigus marmoreus with high activity of ligninolytic enzyme were screened and their biological and genetic characteristics were identified by remazol brilliant blue R-PDA plate chromogenic test, aniline blue-PDA plate decolorization test, somatic cell incompatibility test and ISSR（inter-simple sequence repeat） molecular marker technology, in order to enrich excellent varieties of Hypsizigus marmoreus.The results showed that the ligninolytic enzyme activity of hybrid strain JZB2124031 was the highest.Remazol brilliant blue R-PDA plate and aniline blue-PDA plate could appear obvious discoloration ring and decolorization ring, respectively.It could produce antagonistic reaction with other strains.The 15 ISSR molecular marker primers showed amplification stability.When the similarity coefficient was 0.75,the 22 strains could be divided into nine groups.Among them, the hybrid strain JZB2124031 was the only one group.In conclusion, the hybrid strain JZB2124031 had a high activity of ligninolytic enzyme and great difference in genetic relationship with other test strains.