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The changes of hydrogen peroxide (H2O2) metabolism and antioxidant enzyme activities in a hybrid poplar (Populus simonii ×P. pyramidalis ‘Opera 8277') in response to mechanical damage (MD) and herbivore wounding (HW) were investigated to determine whether H2O2 could function as the secondary messenger in the signaling of systemic resistance. Results show that H2O2 was generated in wounded leaves through MD and HW treatments and systemically in unwounded leaves around the wounded leaves. The activities of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) were also enhanced. However, the H2O2 accumulation and antioxidant enzyme activities were inhibited in MD leaves through the pretreatment with DPI (which is a specific inhibitor of NADPH oxidase). The results of this study suggest that H2O2 could be systemically induced by MD and HW treatments, and H2O2 metabolism was closely related to the change in SOD, APX and CAT activities. A high level of antioxidant enzymes could decrease membrane lipid peroxidation levels and effectively induce plant defense responses.