Leaf morphology and the leaf protein expression profiles of flue-cured tobacco grown in central Henan province of China under low nitrogen (low-N) and normal nitrogen (normal-N) nutrition were examined. The leaf length and width were measured at 50, 60, and 70 days after transplanting. Leaves grown under low-N conditions were shorter and more narrow than those grown under normal-N conditions. The protein expression profiles of tobacco leaves harvested at 70 days after transplanting were analyzed by 2-dimensional electrophoresis, and five differentially expressed proteins including a putative protein were identified. Except for the MCM protein-like protein, the other three differentially expressed proteins of cyclophilin-like protein, vacuolar invertase INV2, MAR-binding protein and the one putative protein showed increased expression in the low-N nutrition group. Among these proteins, the cyclophilin-like protein, which is a stress-responsive signal protein, may play pivotal roles in regulating leaf development under stress conditions. Real-time quantitative PCR analysis showed that the mRNA expression level of the cyclophilin-like protein at day 50, 60 and 70 under low-N conditions was 0.90, 1.43 and 6.9-fold higher than that under normal-N conditions, indicating that the gene expression of cyclophilin-like protein was strongly induced by low-N conditions.