The autoclaving assisted by an irradiation pretreatment method was developed without toxic chemicals to produce fermentable sugars for their conversion to bioethanol. In the first step, electron beam irradiation (EBI) of rice straw was performed at various doses. The electron beam-irradiated rice straw was then autoclaved with DI water at $120^{circ}C$ for 1 h. A total sugar yield of 81% was obtained from 300 kGy electron beam-irradiated rice straw after 72 h of enzymatic hydrolysis by Cellulase 1.5L (70 FPU/mL) and Novozyme-188 (40 CbU/mL). Also, the removal of hemicellulose and lignin was 32.0% and 32.5%, respectively. This result indicates that the environmentally-friendly pretreatment method of rice straw by an electron beam irradiation could be applied for bioethanol production in plant.