Life Cycle Assessment (LCA) is the only standardized tool currently used to assess environmental loads of products and processes. The life cycle analysis, as a part of LCA, is a useful and powerful methodology for studying life cycle energy efficiency and life cycle GHG emission. To quantitatively explain the potential of energy saving and greenhouse gas (GHG) emissions reduction of corn stover-based ethanol, we analyzed life cycle energy consumption and GHG emissions of corn stover-based ethanol by the method of life cycle analysis. The processes are dilute acid prehydrolysis and enzymatic hydrolysis. The functional unit was defined as 1 km distance driven by the vehicle. Results indicated: compared with gasoline, the corn stover-based E100 (100% ethanol) and E10 (a blend of 10% ethanol and 90% gasoline by volume) could reduce life cycle fossil energy consumption by 79.63% and 6.25% respectively, as well as GHG emissions by 53.98% and 6.69%; the fossil energy consumed by biomass stage was 68.3% of total fossil energy input, N-fertilizer and diesel were the main factors which contributed 45.78% and 33.26% to biomass stage; electricity production process contributed 42.06% to the net GHG emissions, the improvement of technology might reduce emissions markedly.