Shii-take cultivation waste (SCW) was used as rich carbon material for anaerobic digestion under the mesospheric condition as (35±1)℃. Orthogonal test and regression analysis L9(33) were employed to determine the effects of the three factors including pretreatment period, inoculums concentration and total solids (TS) on biogas production. The optimal combination of the three factors has been achieved. The high-carbon content material was proved to be not an ideal feedstock for biogas production. However, after a certain period of pretreatment, the high carbon feedstock biogas production was improved a lot. The biogas production reached to 0.16L/g(TS), increased by 128.57% compared with the control sample. The sequence of factors effecting SCW anaerobic fermentation is pretreatment period (evident), TS (evident), and inoculums concentration (less evident). A multiple regression equation was established and can be used for biogas production prediction under different conditions.