Abstract: Adopting mycelial biomass of Lentinus edodes(Berk.) Sing as major index,different carbon and nitrogen source on the L.edodes(‘Nanshan NO.1') mycelial growth were used in the single factor test. The important factors influencing mycelial biomass of L.edodes which identified by initial experimental design of Plackett-Burman were corn flour,wheat bran,Mg SO4·7H2O and inoculation quantity. The path of steepest ascent was undertaken to approach the optimal region of the four significant factors. Box-Behnken design and response surface analysis were adopted to further investigate the mutual interaction between the variables and obtain optimal values that bring the optimal L.edodes liquid seed production process conditions. The optimal conditions for L.edodes liquid seed production were achieved and listed as follows:corn flour 36.0 g,wheat bran 21.0 g,brown sugar 20.0 g,beef powder 4.0 g,KH2PO43.0 g,Mg SO4·7H2O 0.6 g,VB110.0 mg,H2 O 1.0 L,initial p H6.2,inoculation quantity 7.0%,temperature 29 ℃,agitation speed of rotary shaker 180 r/min,culture time 10 days. Under the optimal conditions,the mycelial biomass of L.edodes was 51.004 g/L,compared to the theoretical value,the relative error of 2.098%. So the response surface method was feasible for the optimization of liquid seed production process of L.edodes,and the predicted values of Mathematical model were consistent with experimental observations.