The content of total fermentation materials TS was 8%. The fermentation materials were made up of distiller's grains of multiple-grains and rice straw in different proportion. The daily biogas production,cumulative biogas production,p H value and VFA in the anaerobic fermentation process were analyzed under 30℃. The results showed that the gas production could start faster using distiller's grains alone and reached production peak on the second day of fermentation,but daily gas production declined rapidly after that day. Using the rice straw and distiller's grains as fermentation materials could reduce daily gas production and slow the startup time. In addition,rice straw could extend the fermentation time,in other words,distiller's grains could reduce the fermentation time. Furthermore,among all the groups,the cumulative gas yield in group with distiller's grains and rice straw of 1∶ 2 was the highest,which reached 8 600 m L,moreover,p H value reduction in this group was minimum and the restraint of acidification was the best. The results could provide evidence for characterization of biogas production of rice straw mixed with distiller's grains of multiple-grains in anaerobic fermentation.