The pollution by heavy metals in coastal waters has gradually intensified due to industrial development. In this study, physiological responses of Ulva lactuca, one of the most common green seaweeds with important ecological and economic value in the global intertidal zone, to acute copper stress were investigated. Results showed that an increase in copper ions concentration significantly inhibited photosynthetic activity and inorganic nitrogen utilization by U. lactuca but, increased its respiration. Copper stress limited the activity and gene expression of enzymes related to carbon and nitrogen assimilation of U. lactuca. Under moderate copper stress, U. lactuca had higher soluble carbohydrate and soluble protein contents, whereas under high copper stress, these components decreased sharply. Copper stress increased malonaldehyde content, and relative electrical conductivity in U. lactuca, but changes in antioxidant enzyme activities were not significant and even slightly decreased. Moreover, the contents of 8-hydroxy-deoxyguanosine and polyADP ribose polymerase in U. lactuca increased by high Cu2+ concentration culture, indicating that oxidative damage caused by high Cu2+ level involved its DNA damage and interfered with DNA repair in the alga. Copper stress seemed to be more damaging to the carbon assimilation process of U. lactuca, resulting in weakened resistance to copper stress and lower growth rate. This reflected the threat of coastal high copper stress to intertidal biodiversity. This provided ecological reference for the assessment of offshore heavy metal pollution represented by copper.