In this study, we investigated the performance and elucidated the synergistic effects of microalgae-fungi symbionts co-cultured with 10(-7) and 10(-9) mol L-1 of GR24 and supplemented with endophytic bacteria, multi-walled carbon nanotubes (MWCNTs) or vitamin B12 (VB12), on nutrient removal and biogas upgrading. The results showed that the microalgae-fungi-bacteria symbiotic system co-cultured with 10(-9) mol L-1 GR24 presented the optimal growth performance of 0.368 & PLUSMN; 0.04 d(-1), chlorophyll a of 249.36 & PLUSMN; 22.31 & mu;g L-1, and extracellular carbonic anhydrase activity of 42.55 & PLUSMN; 3.755 enzyme units. In this co-culture system, the organic matter, nutrients, and CO2 purification obtained the highest removal efficiency, with 81.35 & PLUSMN; 7.96% for chemical oxygen demand, 83.56 & PLUSMN; 7.91% total nitrogen, 84.17 & PLUSMN; 7.95% total phosphorus, and 63.72 & PLUSMN; 6.06% CO2. The symbiont system also greatly increased the methane content in the biogas by 30.67%. The remarkable performance of the microalgae-fungi-bacteria symbiotic system shows its ability to be broadly applied in simultaneous biogas upgrading and wastewater treatment. Practitioner PointsThe optimal GR24 concentration for microalgae-fungi consortia was 10(-9) M.Endophytic bacteria were superior to MWCNTs and VB12.Fungi-algae-bacteria consortia presented excellent growth and removal performance.Removal efficiencies of COD, TN, and TP were about 81% under optimum treatment.