By using 104 pairs of SSR primers designed from the whole genome of Lentinula edodes，the genetic diversity of 21 protoplasted monokaryons of Lentinula edodes was analyzed． The results showed that 58 pairs of SSR primers，accounting for 55． 8% of the designed primers，produce 298 polymorphic fragments，accounting for 69． 3% of the total 430 amplified fragments． The genetic similarity among the tested stains was analyzed by using NTSYS-pc21 software，and the results found significant differences existed between the cultivated strain and the wild strain． However，the cultivars with high similarity coefficients exhibited close genetic relationships between each other． At the same time，the study also showed that very close genetic relationship exists between the two protoplast mononuclear strains obtained from the same two nucleosomes．