Copper has been widely used to control algae and pathogens in fish culture ponds. However, its toxic effects on fish depend not only on its concentration in the water but also on the water quality. A laboratory experiment was conducted to assess copper toxicity in the black rockfish Sebastes schlegeli using a panel of antioxidant enzymes, including glutathione (GSH), glutathione S-transferase (GST), glutathione peroxidase (GPx), glutathione reductase (GR) and superoxide dismutase (SOD), at different levels of copper at three water temperatures (WT, 18, 23, 28°C) for 4 days. After exposure to two copper concentrations (100 and 200 μg/L), GSH levels and GST activities increased significantly, depending on WT (P < 0.05) in the liver, gill, and kidney of the black rockfish. GPx and SOD activities decreased significantly with both increasing WT and copper treatment in the organs of black rockfish (P < 0.05). These changes can be seen as initial responses to temperature stress and as a sustained response to copper exposure. This also indicates that GSH and related enzymes activities were sensitive indexes to stress by toxicants such as copper. The present findings suggest that simultaneous stress due to temperature change and copper exposure can accelerate changes in enzymes activities in the black rockfish. This provides another example of synergism between environmental temperature and pollutants, which may have important implications for the survival of fish in polluted environments during seasonal warming and/or global climate change.