Six treated lignocellulosic samples from three feedstocks (switchgrass, corn stover, and anaerobically digested manure fiber) under two pretreatment methods (alkali and acid treatments) were used to culture Trichoderma reesei Rut C-30. Effects of treated lignocellulosic materials on fungal cellulase/xylanase production and corresponding hydrolysis were elucidated. The results demonstrated that the cultures on alkali treated samples had better enzyme production than the acid treated samples. The consequent hydrolysis indicated that with the same feedstock and hydrolysis conditions, enzymes from cultures on different treated samples had significantly different hydrolysis performance compared to commercial enzymes and enzymes from cultures on chemical defined medium. The statistical comparison further demonstrated that enzymes from cultures on alkali and acid treated lignocellulosic materials were favorable to hydrolyze the corresponding groups of the treated materials.