Sanjiang Plain is intensively used for rice production, and ditch drainage diffuse pollution prevention is crucial. Groundwater, rich in Fe ions, is the main source of irrigation water in this region. In this study, pyrite and zero-valent iron (ZVI) (sponge iron and iron scraps) were used as substrates to identify the synergistic influence of exogenous Fe²⁺ addition and solid iron substrates on pollutant removal in constructed wetlands. Based on the results, iron substrates hardly improved the ammonia removal, mainly because of the physical structure and oxidation activity. At a hydraulic retention time longer than 8 h, the pollution removal efficiency in the zero-valent iron (ZVI) substrate treatment increased significantly, and the removal of nitrate (NO₃ ^--N) and total phosphorus (TP) in the iron scrap substrate treatment reached about 60% and 70%, respectively. The high-throughput sequencing results showed a significant increase in the abundance of microorganisms involved in denitrification and phosphate accumulation in biofilms on ZVI substrates. The highest diversities of such microorganisms in biofilms on iron scraps were found for denitrifying bacteria (Pseudomonas), nitrate-reducing Fe (II)-oxidizing bacteria (Acidovorax), and Dechloromonas with autotrophic denitrification and phosphate accumulation, with a 43% cumulative abundance. Dechloromonas dominated in the iron sponge substrate treatment. The highest relative abundance of Acidovorax was found in the mixed iron substrate (pyrite, sponge iron, and iron scraps) treatment. The addition of ZVI substrate significantly improved the removal of NO₃ ^--N and TP and reduced the hydraulic retention time through the continuous release of Fe²⁺ and the promotion of microbial growth. When designing constructed wetlands for treating paddy field drainage, the appropriate addition of iron scrap substrates is recommended to enhance the pollutant removal efficiency and shock load resistance of CWs.

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