Abstract: The microbial fermentation bed humus is the waste produced in the modernization animal husbandry breeding industry. In order to avoid causing the environmental pollution, it must reuse it with multimode as resource recovery. Comparing its mycelium morphology and growth speed, and evaluating the suitability of stropharia rugoso-annulata mycelium culture with microbial fermentation bed humus as substitute material, by microbial fermentation bed humus with different proportion as the main substrate raw material. The results showed that the mycelium of group Ⅰwas gracile and sparse that material wheat bran completely replaced, it was not used as a substitute material; the mycelium of groupⅡ grew slowly that material sawdust completely replaced; the mycelium of the other formula that adding the wheat bran were all pure white and thickening, the growth situation of the mycelium was same as group CK. In view of the same growth situation of the mycelium, it can continue optimizing the application method of microbial fermentation bed humus as a substitute material, and select the best substitution proportion to culture the mycelium of stropharia rugoso-annulata.