The effects of maize variety of Ruifeng 1-double resistance 12-5 (RF1-12-5) on enzyme activity and microbial community in rhizosphere soil were studied to provide scientific safety assessment data for its environmental release and commercial application. The results showed no significant difference existed in the activities of alkali protease, urease, acid invertase between RF1-12-5 and the non-transgenic maize Ruifeng 1 (RF1) in five growth stages of maize. The activities of catalase and alkaline phosphatase were not significant different between RF1-12-5 and RF1 in other growth stages except that catalase activity in harvest stage and alkaline phosphatase activity in milk mature stage were significantly lower for RF1-12-5 than RF1. No significant differences existed in the indexes of Shannon diversity, McIntosh evenness and Simpson dominance of rhizosphere soil microbial communities between RF1-12-5 and RF1 in five growth stages. Principal component analysis proved no regular difference existed between RF1-12-5 and RF1.