Abstract: In order to thoroughly investigate the protein level changes of tall fescue in low-nitrogen conditions, we analyzed the proteome of the leaves from 30 day old plants exposed to low-nitrogen stress using the iTRAP technique. In total, 595 proteins were differentially accumulated (295 up-regulated and 300 down-regulated), which participated in diverse metabolic pathways. According to a strict selection standard, we discovered that the genes related to redox reactions and stresses significantly increased. Physiological and biochemical analysis revealed that the contents of chlorophyll, soluble proteins and free amino acid dramatically decreased, while reactive oxygen-scavenging enzymes such as POD, SOD and GS were highly active. The expression pattern of affected genes, detected by real-time fluorescence quantitative RT-PCR, coincided with the proteomic data. Notably, most of the 14-3-3 family of proteins were enriched by nitrogen shortage, suggesting that these involve the key genes that take part in diverse stresses in tall fescue. In this study, we provide proteomic information for tall fescue subjected to low-nitrogen availability. We also highlight some of the key genes involved and discuss their potential uses.