Abstract: In order to obtain the optimization condition of Trichoderma atroviride Producing the Spore, the factors affecting the sporulation quantity, such as the cultivating temperature，the initial pH, the volume of medium and the volume of inoculum were investigated in present study. The results showed that, the sporulation quantity was best with 0.1 mL inoculum (1.0×107 cfu/mL) in the 20 mL medium under the initial pH 6 of 25℃ for 7 days.