Perennial grasses undergo compensatory growth after defoliation. Nitrate is the main nitrogen source for the growth of perennial ryegrass and plays a significant role in plant resistance to stress. The aim of the study was to understand the physiological mechanism of ryegrass in response to defoliation stress under different nitrate supplies and to explore possible ways to alleviate defoliation stress. We performed pot experiments where 12-week-old ryegrass plants grown in low (0.05 mM KNO₃) or moderate nitrate (5 mM KNO₃) conditions were defoliated and subsequently supplied with different concentrations of nitrate following defoliation treatments. During the regrowth stage, the regrowth rate, biomass, photosynthetic parameters, and the response of the antioxidant system to low or moderate nitrate supply of ryegrass were investigated. The results showed that moderate nitrate supply after defoliation increased the content of photosynthetic pigments in ryegrass and improved its photosynthetic efficiency. In addition, adding moderate nitrate after defoliation increased the activity of antioxidant enzymes and the accumulation of osmotic regulating substances, thereby enhancing plant resistance, effectively reducing the damage to plants caused by defoliation stress, and promoting plant regrowth, especially for plants grown in a low nitrate environment before defoliation. Therefore, this study showed that the addition of exogenous nitrate could counteract some of the adverse effects of defoliation stress on the growth and development of ryegrass.