In Missouri, distinct geophysical gradients influence statewide patterns in water quality. Here, we quantify the spatiotemporal variability of dissolved organic carbon (DOC) in reservoirs and streams and the edaphic, hydrologic, and land cover variables that account for cross-system variation. Datasets included statewide inventories collected over decades and studies with greater temporal resolution (n = >6350 DOC measurements). Among reservoirs, the smallest DOC concentration was measured in a spring-fed system within a forested watershed, and the largest was where agricultural biosolids were applied to the land (range 1.0–15.9 mg/L, overall mean 5.8 mg/L). Reservoir values increased from the southern forested Highlands (mean 4.7 mg/L) to the northern agricultural Plains (mean 7.0 mg/L). Stream DOC was similar to reservoir values (overall mean in streams 6.3 mg/L; Highlands mean 4.0 mg/L; Plains mean 6.6 mg/L), despite differences in study design and collection period. Reservoir DOC increased in spring, indicative of allochthonous loading, with small autochthonous additions during a broad summer peak. Temporal variability in DOC was low relative to macronutrients and chlorophyll in both reservoirs and streams, indicating DOC may be a sensitive and readily detected indicator of temporal change in these systems. In regression analyses, watershed features accounted for more than 60% of overall cross-system variability in DOC in both reservoirs and streams. Driver-response relations, however, differed between regions. This analysis extends our understanding of environmental influences on surface water chemistry in Missouri and indicates DOC is nearly as predictable as macronutrients using landscape-level features.