Interplanting is an efficient method of improving nutrient utilization. However, the impact of intraspecific interplanting on rhizosphere microbial nitrogen cycling needs to be studied further. In this study, two corn cultivars were selected as the materials: Zhengdan958 (ZD958, high nitrogen use efficiency) and Denghai3622 (DH3622, low nitrogen use efficiency). Three planting patterns (interplanting, ZD958 monocropping, and DH3622 monocropping) were set up to study the effects of interplanting on crop growth and rhizosphere microbial nitrogen cycle function under two nitrogen levels: low nitrogen (140 kg N ha−1) and normal nitrogen (280 kg N ha−1). The results showed that the grain yield and nitrogen content in interplanting were significantly increased due to an enhanced leaf area index and root dry weight. The nitrogen accumulation and nitrogen use efficiency were enhanced by 8.14% and 19.38% in interplanting, which resulted in reductions in NH4+ and NO3− content in the rhizosphere. Interplanting enhanced rhizosphere nitrogen cycling processes; nitrification, denitrification, and nitrate reduction were increased. This study demonstrated that interplanting promotes corn nitrogen acquisition from the soil and indirectly regulates rhizosphere microbial function. These findings imply that the intraspecific interplanting of crops with appropriate functional traits is a promising approach to establishing diversified, productive, and efficient resource utilization ecosystems.