Abstract: Objective: Mushroom spawn provided by Zhoukou Academy were tested for its resistance to selenium range.Method: Through single factor experiment and response surface methodology Box- behnken Design ( BBD) optimization test.3, 3'- diaminobenzidine hydrochloride method was applied to determine the selenium content of Mushroom under the conditions of different components and proportion. Result: The results of single factor experiment showed as follows: the optimum value of appropriate selenium content for Mushroom mycelium growth was 300 mg / kg training materials, the best value of wheat bran content was 200 g / kg cottonseed hulls, the optimum value of sucrose content was 12 g / kg cottonseed hulls.The optimum Se Mushroom growth formula was determined by response surface experiment: wheat bran 200.14 g / kg cottonseed hulls, sucrose 11.77 g / kg cottonseed hulls, selenium 318.27 mg / kg training materials.The measured value of selenium content of Mushroom was 2.1859 mg / kg, under the optimal conditions, which was well consistent with the predicted value of 2.1878 mg / kg. Conclusion: This research for the rich selenium Mushroom provided theory basis for the scale of production, and had a certain reference value.