In this study fungal pretreatment of non-sterile tall wheat grass via the white rot fungi Flammulina velutipes was studied and the effect on biodegradability of lignocellulosic biomass and methane production, was evaluated. Degradation of lignin, cellulose, hemicellulose, and dry matter in non-sterile tall wheat grass during 28 days of fungal pretreatment using different inoculum ratio (0%-50%) and moisture content (MC) (45% MC, 65% MC, and 75% MC) were assessed via comparison to untreated biomass. Pretreatment with F. velutipes was most effective at 65% MC and 40% inoculum ratio, resulting in 22% lignin removal. The corresponding methane yields were 181.3 Ndm(3) kg VS-1,which were 280% higher than for the untreated tall wheat grass.