The plant carbon source and sulfur were selected as the denitrification electron donors and filled in the internal water storage zone (IWSZ) of bioretention system to establish excellent mixotrophic denitrification system, which was beneficial to waste recycling and showed very high nitrate nitrogen removal efficiency (approximately 94%). The ammonia nitrogen, total nitrogen, and chemical oxygen demand removal efficiencies could reach 79.41%, 85.89%, and 74.07%, respectively. Mechanism study revealed the synergistic degradation effect was existed between acetic acid released from plant carbon source and the generated sulfate, which improved the S/CH3COOH mediated nitrate nitrogen removal reactions. Autotrophic denitrification occurred mainly in the upper layer of IWSZ, and the dominant bacteria were Thiobacillus. While in the lower layer, the dominant bacteria were mainly related to organic matter utilization and heterotrophic denitrification. The abundance of narG, nirK, nirS, and nosZ functional genes in the upper layer was significantly higher than the lower layers.