[Objective]The paper was to study the effect of different nitrogen application levels and modes on contents of different forms of nitrogen in rice plants.[Method] Using Guichao 2 and Diejiazhan as test materials,with total nitrogen,protein nitrogen,chlorophyll nitrogen,soluble protein nitrogen,Rubisco nitrogen,free amino acid nitrogen as indicators,the effect of nitrogen application on contents of different forms of nitrogen in rice plants were studied.[Result] The contents of total nitrogen,protein nitrogen,chlorophyll nitrogen,soluble protein nitrogen,Rubisco-N,free amino nitrogen of rice plant during heading and maturity stage increased with the increasing application amount of nitrogen,and their contents would be high under heavy application of spike fertilizer when the application amount of nitrogen was the same.However,application of nitrogen had different impact on different forms of nitrogen,which showed obvious impact on the contents of Rubisco-N and free amino nitrogen.The differences of Rubisco-N and free amino nitrogen content of two varieties during heading and maturity stage under different nitrogen application modes all reached significant or extremely significant level.Correlation analysis showed that total nitrogen content of rice plant had extremely significant correlation with protein nitrogen content during heading and maturity stage in both early and late seasons.[Conclusion] The study provided a theoretical basis for clarifying the regulatory role of nitrogen application on nitrogen absorption and utilization.