Abstract: The spores of Ganoderma lucidum are difficult to germinate under the artificial conditions ,so it’s unable to get the monokaryon strains which is necessary in breeding of Ganoderma lucidum, and the advancement (progress )of breeding is slower. In this study, the statistical software and molecular biology methods were used to select the parents with genetic distance distant and complementary agronomic characters. In this way, the G.l0006, G.l0012, G.l0024, G.l0030+2, and G.l0003+1 were screened out as the hybrid parent strains. 81 mononuclear strains were obtained through protoplasted monokaryon technique, and they were used as hybrid breeding strains, 24 hybrids were obtained. The hybrids were verified to be different from their parents by antagonist response.