Propachlor is a widely used acylaniline herbicide for weeding annual gramineous and dicot plants in farmland. As a toxic agricultural chemical, it is overused in crop production and has become one of the most serious environmental pollutants. Yet, understanding the impact of environmental factors on its behavior in farmland soils is critically important for healthy crop production and food safety. In this study, we characterized the effect of dissolved organic matter (DOM) extracted from rice straw on the mobility behavior of propachlor in farmland soil using comprehensive approaches such as the batch equilibrium experiment, soil thin-layer chromatography, soil column leaching, and wheat bioaccumulation with Triton X-100 (TX-100) as a reference surfactant. The application of DOM at 60 and 120 mg DOC L–1 and TX-100 at 96, 192, and 288 mg L–1 reduced the sorption and increased the desorption of propachlor in soil. Freundlich constant K f values (sorption) of propachlor declined with the expansion of DOM and TX-100 concentrations. The addition of DOM and TX-100 increased the mobility of propachlor and the total concentration of propachlor in the leachate of soil columns. The retention factors (R f) were evaluated in the soil thin-layer chromatography as 0.741 and 0.772 (for DOM) and 0.731, 0.763, and 0.791 (for TX-100), all of which were greater than the control (0.710) under the treatment. The application of DOM or TX-100 reduced root growth (biomass) and increased bioaccumulation of propachlor in the roots and shoots of wheat. The outcome of the study can provide important references for the rational use of propachlor and help agronomic management to minimize pesticide contamination in realistic crop production.