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As the biogeochemical cycling of S in soil is closely associated with the mobility and bioavailability of heavy metals, in this study, we investigated the influence of S fertilization (S and SO) in paddy soils on rice growth, iron plaque formation over root surface, and Cu speciation of rice rhizosphere soil under flooding conditions. The dry weight and height of rice plants increased significantly after S fertilization, indicating that S fertilization of Cu-contaminated paddy soils can enhance rice growth. Sulfur fertilization (less than 500 mg/kg) promoted the formation of iron plaque, thus sequestering a large amount of Cu on root surface and decreasing the bioavailability of Cu by inducing transformation of Cu bioavailable fractions (exchangeable, carbonate oxides, or iron and manganese oxide bound to Cu) to Cu bound to organic matter. Copper K-edge X-ray absorption near-edge structure (XANES) revealed that S fertilization increased the percentage of Cu present as CuS and Cu-cysteine in rice rhizosphere soil, thus reducing Cu mobility. As Cu concentration in rice plants decreased and the biomass of rice plants increased after S fertilization, it can be suggested that S fertilization may be an effective approach for managing Cu-contaminated paddy soils. [ABSTRACT FROM AUTHOR]