Abstract: The yields of pinoresinol diglucoside（ PDG） and cordycepin in the fermentation products using Eucommiae cortex as medicinal substrate by Cordyceps militaris（ L.） Link. were improved，and optimal conditions for fermentation production were obtained. Factors affecting cordycepin and PDG production in the fermentation were screened by applying the Plackett-Burman design by the pre-experiment，and the conditions of cordycepin and PDG production were further optimized by Box-Benhnken design. The results of Plackett-Burman experiment showed that these factors of the amount of additive，the cultivation time and ammonium sulfate had notably influenced the transformation of E. cortex in solid state fermentation by C. militaris. The conditions of cordycepin and PDG productionwere further optimized by Box-Behnken design. The results were as follows: VE，glucose，ammonium sulfate，initial pH，additive，inoculation amount，cultivation temperature and cultivation time are respectively 4 g/kg，4 g/kg，8 g/kg，8，27%，40%，25℃ and 18 d. Cordycepin and PDG production in the solid state fermentation had reached up to 2.92 μg/g and 90.03 μg/g respectively，which were 60.44% and 52.90% increase than the initial value by this optimal process. This study indicated that the optimization design proved a useful and powerful tool for the development of optimal medium compositions and culture conditions. C. militaris strain B1 528 was a promising fungus strain for PDG and cordycepin production in solid state fermentation using E. cortex as medicinal substrate.