Accurate nitrogen (N) nutrition diagnosis is essential for improving N use efficiency in crop production. The widely used critical N (Nc) dilution curve traditionally depends solely on agronomic variables, neglecting crop water status. With three-year field experiments with winter wheat, encompassing two irrigation levels (rainfed and irrigation at jointing and anthesis) and three N levels (0, 180, and 270 kg ha−1), this study aims to establish a novel approach for determining the Nc dilution curve based on crop cumulative transpiration (T), providing a comprehensive analysis of the interaction between N and water availability. The Nc curves derived from both crop dry matter (DM) and T demonstrated N concentration dilution under different conditions with different parameters. The equation Nc = 6.43T−0.24 established a consistent relationship across varying irrigation regimes. Independent test results indicated that the nitrogen nutrition index (NNI), calculated from this curve, effectively identifies and quantifies the two sources of N deficiency: insufficient N supply in the soil and insufficient soil water concentration leading to decreased N availability for root absorption. Additionally, the NNI calculated from the Nc-DM and Nc-T curves exhibited a strong negative correlation with accumulated N deficit (Nand) and a positive correlation with relative grain yield (RGY). The NNI derived from the Nc-T curve outperformed the NNI derived from the Nc-DM curve concerning its relationship with Nand and RGY, as indicated by larger R2 values and smaller AIC. The novel Nc curve based on T serves as an effective diagnostic tool for assessing winter wheat N status, predicting grain yield, and optimizing N fertilizer management across varying irrigation conditions. These findings would provide new insights and methods to improve the simulations of water-N interaction relationship in crop growth models.
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