Sisal leaf decortications residue (SLDR) is amongst the most abundant agroindustrial residues in Tanzania and is a good feedstock for biogas production. Pretreatment of the residue prior to its anaerobic digestion (AD) was investigated using a twostage pre-treatment approach with two fungal strains, CCHT-1 and Trichoderma reesei in succession in anaerobic batch bioreactors. AD of the pre-treated residue with CCTH-1 at 10% (wet weight inoculum/SLDR) inoculum concentration incubated for four days followed by incubation for eight days with 25% (wet weight inoculum/SLDR) of T. reesei gave a methane yield of 0.292 ± 0.04 m3 CH4/kg volatile solids (VS)added. On reversing the pre-treatment succession of the fungal inocula using the same parameters followed by AD, methane yield decreased by about 55%. Generally, an increment in the range of 30–101% in methane yield in comparison to the un-treated SLDR was obtained. The results confirmed the potential of CCHT-1 followed by Trichoderma reesei fungi pre-treatment prior to AD to achieve significant improvement in biogas production from SLDR.