We investigated the effects of increasing soil NaCl concentration on intracellular compartmentalization of salt and on the activities of antioxidant enzymes (superoxide dismutase (SOD), ascorbic peroxidase (APX), catalase (CAT) and glutathione reductase (GR)) and their role in the regulation of reactive oxygen species (ROS; O(2)(-*) and H(2)O(2)) in leaves and xylem sap of salt-tolerant Populus euphratica Oliv. and salt-sensitive P. popularis cv. 35-44. Mesophyll cells of P. euphratica exhibited a high capacity for NaCl exclusion and compartmentalization of salt in vacuoles compared with P. popularis. In P. popularis, the salt treatment resulted in large accumulations of Na(+) and Cl(-) in leaves that induced significant increases in O(2)(-*) and H(2)O(2) production despite marked increases in the activities of antioxidant enzymes in leaves and xylem sap. Separation of the isoforms of leaf SOD, APX and CAT by polyacrylamide gel electrophoresis followed by in-gel activity staining revealed that the salt-induced activities of APX and CAT were the result of increases in activities of all the isoenzymes. Leaf injury and shedding of aged leaves occurred following the oxidative burst in P. popularis, indicating that the increased activities of antioxidant enzymes in P. popularis were insufficient to counter the harmful effects of ROS at high soil NaCl concentrations. Unlike P. popularis plants, P. euphratica plants did not exhibit an oxidative burst in response to the NaCl treatments, because of (1) a high salt exclusion capacity and effective compartmentalization of salt in vacuoles, and (2) up-regulation of antioxidant enzymatic activities after the onset of salt stress. We conclude that P. euphratica plants subjected to saline conditions control ROS homeostasis through two pathways: (1) by maintaining cellular ionic homeostasis and thereby limiting the NaCl-induced enhancement of ROS production under long-term saline conditions; and (2) by rapidly up-regulating antioxidant defenses to prevent oxidative damage.