Glufosinate-ammonium (GLAM) can be released into adjacent water bodies with rainfall runoff and return water from farmland irrigation. However, impacts of GLAM on aquatic organisms remain unclear. In this study, changes in water quality, plant physiological parameters and epiphytic microbial community were investigated in wetlands with Hydrilla verticillata exposed to GLAM for 24 days. We found GLAM addition damaged cell and reduced chlorophyll a content in Hydrilla verticillata leaves, and increased ammonium and phosphorus in water (p < 0.001). The α-diversity increased in bacterial community but decreased in eukaryotic community with GLAM exposure. Neutral community models explained 62.3 % and 55.0 % of the variance in bacterial and eukaryotic communities, respectively. Many GLAM micro-biomarkers were obtained, including some clades from Proteobacteria, Bacteroidete, Actinobacteriota, Phragmoplastophyta, Annelida and Arthropoda. Redundancy analysis revealed that GLAM concentration was positively correlated to Flavobacterium, Gomphonema and Closterium but negatively to Methyloglobulus and Methylocystis. Network analysis revealed that 15 mg/L GLAM disturbed the interactions among phytoplankton, protozoa, metazoan and bacteria and reduced the stability of the microbial communities compared to 8 mg/L GLAM. GLAM shaped the nitrogen and phosphorus cycle related bacterial genes. This study highlights that herbicides are non-neglectable factors affecting the efficiency of aquatic ecological restoration in agricultural areas to control agricultural non-point source pollution.