A novel pretreatment strategy based on combination of microwave and ionic liquid [TBA][OH] was developed for enhancing enzymatic hydrolysis of Eucalyptus sawdust. The sugar yield of pretreated sample achieved 410.67 mg/g in 48 h, which suffered from optimized microwave-assisted [TBA][OH] pretreatment. The work mechanism was illuminated by chemical composition, Fourier transform infrared spectroscopy (FTIR), C-13 cross polarization/magic-angle spinning solid state NMR (C-13 solid NMR), X-ray diffraction (XRD) and scanning electron microscope (SEM) analyses. The combined effect of microwave and [TBA][OH] leads to the violent deconstruction of lignin, removal of hemicelluloses, destruction of crystalline region and an eroded, pored and irregular micro-morphology. As a green, relatively inexpensive and high efficient pretreatment, microwaveassisted [TBA][OH] pretreatment has great potential in the field of bio-refinery.