Per- and polyfluoroalkyl substances (PFAS) are widespread pollutants that can influence microorganisms. To unveil the effects of PFAS in natural microecosystems, a study that focused on the bacterial, fungal, and microeukaryotic communities around the PFAS point source was conducted in China. A total of 255 specific taxa were significantly different between the upstream and downstream samples, 54 of which were directly correlated with PFAS concentration. Stenotrophomonas (99.2 %), Ralstonia (90.7 %), Phoma (21.9 %), and Alternaria (97.6 %) were the dominant genera in sediment samples from the downstream communities. In addition, most of the dominant taxa were significantly correlated with PFAS concentration. Furthermore, the type of microorganism (bacteria, fungi, and microeukaryotes) and habitat (sediment or pelagic) also influence the microbial community responses to PFAS exposure. Pelagic microorganisms featured more PFAS-correlated biomarker taxa (36 pelagic microeukaryotic biomarkers and 8 pelagic bacteria biomarkers) than the sediments (9 sediment fungi biomarkers and 5 sediment bacteria biomarker). In general, around the factory, the microbial community was more variable in pelagic, summer, and microeukaryotic conditions than in other types. Attention needs to be paid to these variables in the future effect of PFAS on microorganisms.