Abstract: Taking M.sextelata strain GZCC0287 as the test material, carbon source, nitrogen source, inorganic salt and sodium carboxymethylcellulose（CMC-Na） were investigate by single factor design, Plackett-Burman design, and central composite design for biomass weight and concentration of mycelium pellet, respectively, in order to provide an effective way for produce the liquid spawns, solid strain and industrial fermentation.The results showed that brown sugar, yeast powder, KH₂PO₄,MgCl₂·6 H₂O were optimal carbon source,nitrogen source and inorganic salt individually.And the optimized medium combination was brown sugar 50.49 g·L^-1,yeast powder 10 g·L^-1,KH₂PO₄1 g·L^-1,MgCl₂·6 H₂O 2.37 g·L^-1,CMC-Na 10.11 g·L^-1.Under this condition,the maximum biomass weight was（5.21±0.45）g and the concentration of mycelia pellets was 2 567±127 per milliliter.The mycelium of Morchella spp.filled the cultivation bag for 10 days to 15 days by inoculated liquid seed 5 mL for 1 000 per milliliter.