Abstract: This study aimed to obtain Lyophyllum decastes mycelium enriching selenium. At the same time, influences of sterilization condition, incubation time, selenium addition amount and time on the growth and efficacy of enriching selenium of Lyophyllum decastes mycelium were investigated by use of dry weight of Mycelia and selenium content as evaluation indicators. The conditions of Lyophyllum decastes mycelium enriching selenium were optimized according to the orthogonal experimental design L9 (34) . The results showed that under the optimum conditions (sterilization for 20min at 115℃, addition of 4μg/mL Na2SeO3 at the 5th day of fermentation, 13 days of fermentation) , dry weight and selenium content of Mycelia respectively reached 0. 8232g/100mL, 0. 7040mg/g. Therefore, the above conditions were the optimum conditions of generating Lyophyllum decastes Mycelium enriching selenium.