In order to investigate the feasibility of using rice leaf critical nitrogen concentration as a nitrogen nutrition diagnosis index, a three-year positioning experiment with large-spike rice cultivar (Wuyoudao4) and multiple-spike rice cultivar (Songjing9) under five nitrogen levels (0, 60, 120, 180, and 240 kg·ha−1) was conducted. A critical nitrogen dilution curve and a nitrogen nutrition index (NNI) of rice leaf dry matter were constructed for Wuyoudao4 (Nc = 1.96LDM−0.56, R2 = 0.87, NNI was between 0.6–1.26, and Normalized Root Mean Square Error (n-RMSE) = 13.07%) and Songjing9 (Nc = 1.99LDM−0.44, R2 = 0.94, NNI was between 0.64–1.29, and n-RMSE = 15.89%). The relationship between dry matter and nitrogen concentration of rice leaves was a negative power function, and the model had good stability over the three years. The developed critical nitrogen concentration dilution curve, based on leaf dry matter, was able to diagnose nitrogen nutrition in rice efficiently. The model established in this study could be used to directly regulate and control the nitrogen nutrition of rice leaves.