Abstract: Paecilomyces fumosoroseus strain IfB01 was respectively cultured in four single medium （rice， soybean powder， corn powder and wheatmeal） and 3 kinds of composite culture medium（wheat flour∶wheat bran = 1∶ 1， soybean flour∶wheat bran = 1∶1 and corn flour∶wheat bran = 1∶1）at 25℃ and RH85% . Using sporulation as the index，better basal culture medium formula was selected. Through orthogonal experiment，adding different concentrations of carbon source and nitrogen source to optimize basal medium，better solid culture medium formula was as follows： corn powder basal medium +0.2% KNO3+0.1% glucose + water（material∶water = 1∶1.5）. Through single factor experiment，the optimum culture conditions of the strain were as follows：at 28℃ and 14h∶10h（l∶d），the humidity was 80% at early stage of spores production，and the humidity was 50% at late stage of spores production.