The global spread of antibiotic resistance genes (ARGs) poses a critical threat to public health and environmental safety. Among environmental factors, the widespread use of chiral pesticides has raised ecological concerns, yet their enantioselective impacts on ARGs propagation remain largely unexplored. Here, we investigate for the first time how chiral pesticides influence microbial ARGs dissemination at the enantiomeric level. Using flurtamone as a model, we successfully separated and quantitatively analyzed its enantiomers (R-flurtamone and S-flurtamone) and evaluated their effects at environmentally relevant concentrations (0-80 μg/L). Remarkably, R-flurtamone significantly enhanced the horizontal transfer of ARGs, surpassing the effects of Rac-flurtamone, whereas S-flurtamone exerted negligible influence. Mechanistic insights revealed that R-flurtamone directly interacts with cell membranes, reducing lipid mobility and increasing permeability, thereby facilitating ARGs spread. Additionally, R-flurtamone induced excessive reactive oxygen species (ROS) production, SOS responses, and boosted ATP levels, further accelerating ARGs propagation. By integrating experimental findings with molecular simulations, we elucidated the enantioselective mechanisms underpinning ARGs transfer. This study highlights the overlooked risks associated with racemic chiral pesticides at enantiomeric level and provides a foundation for mitigating ARGs dissemination in agricultural and environmental systems.