Nitrogen (N) plays crucial roles in chlorophyll concentration, photosynthesis, and stress tolerance of plant leaves. This study conducted a greenhouse experiment combined with Cd and N treatments to elucidate the mechanism underlying the influence of N on Cd accumulation and acclimation strategy in Populus leaves. Chlorophyll concentration and net photosynthetic rates (A) in leaves were unaltered by Cd exposure regardless of N condition. Nitrogen availability alter acclimation strategy of poplar leaves under cadmium exposure. Under sufficient N, Cd accumulation in leaves was elevated with increased intensity and duration of Cd exposure; Cd accumulation reached ca. 28 μg g −1 dry weight and 260 μg plant −1 after 60 days of exposure to high level of Cd (20 mg Cd kg −1 soil), and this finding indicates a large potential for Cd phytoextraction. Poplar leaves exhibited high capacity for antioxidant defense and stress tolerance and avoided oxidative damage under high Cd exposure. The levels of phytohormones and antioxidants in leaves and the relative expressions of critical genes encoding antioxidant enzymes were up-regulated under sufficient N condition. Nitrogen deficiency decreased chlorophyll concentration and net photosynthetic rates (A) and interfered with the production of N metabolites, resulting in a low level of phytohormones and antioxidants that are responsible for stress tolerance. The low levels of Cd accumulation in leaves may be a self-protecting strategy to prevent severe oxidative damage due to the decreased capacities for stress tolerance under N deficiency.