Incorporation of organic fertilizers/amendments in soils has been, and will continue to be, a popular strategy for improving the quality of arable soils. However, the mechanism by which the dissolved organic matters (DOMs) affect soil properties or interact with other substances in soils is largely unknown. In this study, a batch equilibrium experiment was performed to evaluate the effect of two types of DOMs on the behavior of prometryne (a herbicide) in soils. Two sorts of DOMs were derived from lakebed sludge (SL) and rice straw (ST), respectively. The results show that sorption capacity in one soil for prometryne was significantly reduced by application of DOMs, whereas desorption of prometryne was promoted by DOM treatments. To understand the mobility behavior of prometryne in soils with DOMs, a column leaching experiment was carried out. It is shown that both DOMs enhanced the solubility and migration of prometryne in soils. To confirm the role of DOMs in regulating the mobility of prometryne in soils, a soil thin-layer chromatography was performed. The migration of prometryne was promoted by DOMs, which were used as a developing solvent or directly incorporated in the soil thin layer. The data indicate that DOM extracts can modify the mobility of prometryne in soil.