In this paper, the varieties of thin-bark pepper (Capsicum annuum) and line pepper (Capsicum frutescens) were used as the experimental materials, and a pot experiment was conducted in order to explore the effects of root interaction between the same and different pepper varieties on nitrogen (N) uptake and accumulation by analyzing the yield, biomass, nitrogen accumulation of pepper, as well as nitrogen form and content changes in rhizosphere soils under individual or interplanting (cross and parallel) planting modes. The results showed that the dry weights of stems and leaves, pepper yield and fruit N accumulation were higher in the interplanting mode than those of the two individual planting modes, which were meanly increased by 26.67%, 63.65% and 15.08% at fruit stage, respectively. In addition, the pepper yield and fruit N accumulation of parallel planting were the highest, but there was no significant difference in root dry weights among different planting modes at fruit stage. At fruit stage, rhizosphere ammonium/nitrate ratio in interplanting mode was significantly lower than those of the individual planting modes, meanly decreased by 67.39%. Interplanting mode of pepper, especially parallel planting could increase the pepper yield and improve N accumulation, which has a certain feasibility and promotion prospect in the actual vegetable production.