Maize/peanut intercropping system shows the significant yield advantage. Soil microbes play major roles in soil nutrient cycling and were affected by intercropping plants. This experiment was carried out to evaluate the changing of rhizosphere microbial community composition, and the relationship between microbial community and soil enzymatic activities, soil nutrients in maize/peanut intercropping system under the following three treatments: maize(Zea mays L.) and peanut(Arachis hypogaea L.) were intercropped without any separation(NS), by half separation(HS) using a nylon net(50 μm) and complete separation(CS) by using a plastic sheet, respectively. The soil microbial communities were assessed by phospholipid fatty acid(PLFA). We found that soil available nutrients(available nitrogen(Avail N) and available phosphorus(Avail P)) and enzymatic activities(soil urase and phosphomonoesterase) in both crops were improved in NS and HS treatments as compared to CS. Both bacterial and fungal biomasses in both crops were increased in NS followed by HS. Furthermore, Gram-positive bacteria( G+) in maize soils were significant higher in NS and HS than CS, while the Gram-negative(G–) was significant higher in peanut soil. The ratio of normal saturated to monounsaturated PLFAs was significantly higher in rhizosphere of peanut under CS treatment than in any other treatments, which is an indicator of nutrient stress. Redundancy analysis and cluster analysis of PLFA showed rhizospheric microbial community of NS and HS of both plants tended to be consistent. The urase and Avail N were higher in NS and HS of both plants and positively correlated with bacteria, fungi(F) and total PLFAs, while negatively correlated with G+/G– and NS/MS. The findings suggest that belowground interactions in maize/peanut intercropping system play important roles in changing the soil microbial composition and the dominant microbial species, which was closely related with the improving of soil available nutrients(N and P) and enzymatic activities. Soil microbes play major roles in soil nutrient cycling and were affected by intercropping plants. This experiment was carried out to evaluate the changing of rhizosphere microbial community composition, and the relationship between microbial community and soil enzymatic activities, soil nutrients in maize / peanut intercropping system under the following three treatments: maize (Zea mays L.) and peanut (Arachis hypogaea L.) were intercropped without any separation (NS) (50 μm) and complete separation (CS) by using a plastic sheet, respectively. The soil microbial communities were assessed by phospholipid fatty acid (PLFA). We found that soil available nutrients (available nitrogen (Avail N) and available phosphorus Avail P)) and enzymatic activities (soil urase and phosphomonoesterase) in both crops were improved in NS and HS treatments as compared to CS. Both bacterial and fungal biomasses in both crops were increased in NS followed by HS. Furthermore, Gram-positive bacteria (G +) in maize soils were significantly higher in NS and HS than CS, while the Gram-negative (G-) was significantly higher in peanut soil. The ratio of normal saturated to monounsaturated PLFAs was significantly higher in rhizosphere of peanut under CS treatment than in any other treatments, which is an indicator of nutrient stress. Redundancy analysis and cluster analysis of PLFA showed rhizospheric microbial community of NS and HS of both plants The urase and Avail N were higher in NS and HS of both plants and positively correlated with bacteria, fungi (F) and total PLFAs, while negatively correlated with G + / G- and NS / MS. The findings suggest that belowground interactions in maize / peanut intercropping system play important roles in changing the soil microbial composition and the dominant microbial species, which was closely related with the improving of soil availabl e nutrients (N and P) and enzymatic activities.