Agricultural management practices promote organic matter (OM) turnover and thus alter both the processing of dissolved organic matter (DOM) in soils and presumably also the export of DOM to headwater streams, which intimately connect the terrestrial with the aquatic environment. Size-exclusion chromatography, in combination with absorbance and emission matrix fluorometry, was applied to assess how agricultural land use alters the amount and composition of DOM, as well as dissolved organic nitrogen (DON) forms in headwater streams, including temporal variations, in a temperate region of NE Germany. By comparing six agriculturally and six forest-impacted headwater streams, we demonstrated that agriculture promotes increased DOC and DON concentrations, entailing an even more pronounced effect on DON. The major part of DOC and DON in agricultural and forest reference streams is exported in the form of humic-like material with high molecular weight, which indicates terrestrial, i.e., allochthonous sources. As an obvious difference in agricultural streams, the contribution of DOC and particularly DON occurring in the form of nonhumic high-molecular-weight, presumably proteinous material is clearly elevated. Altogether, DOM in agricultural headwaters is mainly complex-soil-derived and aromatic material with a low C:N ratio, which is more microbial processed than its counterpart from forest reference catchments. Our results emphasize the importance of agricultural land use on DOM loss from soils and identify agricultural soils as important DOC and particularly DON sources to headwater streams.