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Constructed wetland-microbial fuel cell (CW-MFC) is a novel technology for wastewater treatment with electrical generation. This work proposed a Fe–C micro-electrolysis substrate (Fe–C) with biomass modified ceramsite to enhance pollutants removal and electricity generation. The key influencing factors were revealed, and the COD, NH 4 +-N, and TP removal efficiency was respectively increased by 10.2, 8.1 and 8.78% with 76% higher power output at optimal conditions (e.g. OLR 52.5 g/(m2.d), HRT 48 h, and aeration rate 800 mL/min). Fe–C based substrates improved the microenvironments in CW-MFC, including dissolved oxygen (DO) and oxidation-reduction potential (ORP) lowering and electron transfer facilitation. These contributed to the enrichment of critical microorganisms and metabolic activities. The abundance of functional bacteria (i.e. Geobacter , Thauera and Dechloromonas) were evidently increased. Additionally, the energy metabolism and other functional genes encoding cytochrome c (ccoN), nitrite reductase (nirD) and phosphate transporter (pstA) were all stimulated. [Display omitted] • Fe–C along with self-developed BMC were applied to the CW-MFC. • Pollutant removal and power generation performance were significantly improved. • Microorganisms were enriched by Fe–C, especial for Geobacter , Thauera etc. • Energy metabolism and other functional genes (ccoN, nirD, and pstA) were stimulated. • The synergism among different particles in the CW-MFC system was discussed. [ABSTRACT FROM AUTHOR]