The use of suitable plants that can accumulate excess phosphorus (P) from contaminated soil may serve as an attractive method for phytoremediation. In this study, pot experiments were conducted to investigate the effects of P incorporation on P accumulation and physiological mechanisms of Polygonum hydropiper in a mining ecotype (ME) and nonmining ecotype (NME) from a phosphorus mining and a noncontaminated agricultural area, respectively. The results demonstrate that the ME of P. hydropiper growing in soil supplied with 0, 100, 200, 400, 800, 1600 mg P (kg soil)–1 showed a significantly higher biomass compared to the NME. Phosphorus accumulation of the ME was positively correlated with the soil P concentration. APase activity in roots of the ME significantly increased at 1600 mg P (kg soil)–1 and phytase activity of the ME increased with increasing P supply. APase activity of the ME was more than twice that of the NME on average. A significant increase of superoxide dismutase (SOD) was observed compared with the NME at all supplied P levels. Peroxidase (POD) activity of the ME was significantly higher at 200 and 400 mg P (kg soil)–1. No statistical differences in the catalase (CAT) activity of the ME were observed compared with the control. Activity of CAT in the NME was obviously induced after exposure to 100–800 mg P (kg soil)–1. Malondialdehyde (MDA) concentration in leaves of the ME decreased with increasing P supply to reach a minimum at 400 mg P (kg soil)–1. In the NME, an increase in MDA concentration compared to the control was observed at higher P levels. The APase and phytase induction and antioxidative defense allowed for the high P accumulation of the ME.