A field experiment was conducted to study the influence of nitrogen fertilization on nitrogen balance index (NBI) and yield of hybrid super rice and evaluate NBI for the assessment of rice nitrogen status and yield. Rice cultivars included two hybrid super rice cultivars (Yongyou12, Dongzhi14) and one common variety (Xiushui 134), nitrogen fertilizer was applied at four levels (0, 200, 300 and 400 kg/hm²). The results showed that the yields of Yongyou 12 and Dongzhi 14 were higher than that of Xiushui 134 at the same amount of nitrogen application, and reached the peak (13.48 t/hm² and 11.51 t/hm², respectively) at N 300 kg/hm², while the yield of common rice reached the peak of 9.49 t/hm² at N 200 kg/hm². Nitrogen fertilizer increased Chl index and NBI but decreased Flav index of Yongyou12 and Dongzhi14. At the heading stage, values of Chl index and NBI of Yongyou12 and Dongzhi14 were significantly higher than that of Xiushui 134. At the tillering and jointing stages, NBI was significantly correlated with leaf nitrogen concentration, rice N accumulation in above-ground and yield. These results demonstrate NBI could be used for rapid nitrogen status diagnosis and yield prediction of rice.