In floodplain rivers, variability in hydraulic connectivity interacts with biogeochemistry to determine the distribution of suspended and dissolved substances. Nutrient, chlorophyll a, and suspended solids data spanning longitudinal (5 study reaches across 1300 river km), lateral (main channel and backwaters), and temporal (1994–2011) gradients in the Upper Mississippi River (UMR) were used to examine the extent to which observed differences between the main channel and backwaters were consistent with expectations based on current understanding of biogeochemical processes in large rivers. For N and P, the results largely conformed to expectations. N concentrations were greater in the main channel than in the backwaters in 82 to 96% of the observations across river reaches. Maximum TP concentrations generally occurred in backwaters during summer, when backwater TP often exceeded that of the main channel. Flux of P from sediments may be a substantial source of water-column P in UMR backwaters in summer. The data for suspended solids and chlorophyll a suggest that some refinements are needed of our understanding of ecosystem processes in large rivers. During low-discharge conditions, concentrations of inorganic suspended solids often were greater in backwaters than in the main channel, suggesting the importance of sediment resuspension. Chlorophyll a concentrations were usually greater in backwaters than in the main channel, but exceptions indicate that phytoplankton abundance in the main channel of the UMR can sometimes be greater than is typically expected for large rivers.