The purpose of this study was to investigate the optimal mixture ratio for the mycelial culture of the Pleurotus ostreatus. The chief cultural media in this study were cotton hull, sawdust and rice straw and the supplemental media were zeolite, corn cob, defatted rice bran, white cotton, tobacco trash powder, rice hull and peat. The results of this study were as follows; the optimal mixture ratio of the chief cultural media were effective in 6 : 3 : 1(V/V, %), and the mycelial growth and density in the supplemental media were considerably better 1% zeolite,3% corn cob, 5% defatted rice bran, 1% white cotton, 1% tobacco trash powder, 7% rice hull in good order. The optimal mixture ratio be to the mixed supplemental media in the chief cultural media were as follows ; 2 : 2(V/V, %) at the conditions of mixed zeolite and corn cob; 3 : 2(V/V, %) at the conditions of mixed defatted rice bran and white cotton; 1 : 3(V/V, %) at the conditions of mixed tobacco trash powder and rice hull. At the conditions of the whole cultural media mixed, the mycelial growth and density were in good conditions ; cotton hull, sawdust, rice straw, zeolite, corn cob, defatted rice bran, white cotton, tobacco trash powder, rice hull, and peat were mixed 43.0 : 17.2 : 25.8 : 2.0 : 2.0 : 3.0 : 2.0 : 1.0 : 3.0 : 1.0 (V/V, %).