Abstract: To set up the technology platform for the crossbreeding of Agaricus bisporus wild strains, and screen out new hybrid cultivars with strong adaptability, high production and good quality, using the crossbreeding technology of Agaricus bisporus homokaryons, the hybridizations between domestic and foreign wild strains, wild strains and cultivation strains were carried out. The results showed that, the proportion of sterile homokaryons in Agaricus burnettii monospore isolates was the highest, up to 92.6%. It was the lowest in traditional cultivation strain, only 8.7%. The colony shape of homokaryons made much difference, and the homokaryons derived from hybrid strain diversified more than those from traditional strains. The homokaryons separated from Sichuan wild strains could be intercrossed with those from foreign wild strains or domestic cultivation strains, further indicating the wild strains from Sichuan to be Agaricus bisporus new germplasm in Tibet altiplano area of China.