The Cinaruco River, an oligotrophic ecosystem in the Venezuelan llanos (savanna), has strong seasonal hydrology and supports large populations of ecologically diverse fishes. The relative contributions of autochthonous and allochthonous production sources that support high stocks of secondary consumers are undetermined in this river. We used excitation–emission fluorescence spectroscopy and absorption spectra of dissolved organic matter to infer degradation of leaf material originating from the surrounding gallery forest. During the low-water period, a large fraction of fluorescent organic matter contained in leaves degraded quickly in river water and was an important allochthonous contribution of C to the system. However, the fluorescence signature of dissolved organic matter in lagoons was different from that of the main river channel during the falling-water period, suggesting that other sources of C were present. Allochthonous organic matter clearly fueled microbial respiration during the falling-water period, but our in-water experiments using light–dark bottle methods indicated that autochthonous production was an important supplementary C source in shallow nearshore waters. During the low-water and falling-water periods, water-column primary production in nearshore waters ranged from 150 to 500 mg C m⁻² d⁻¹ and was 2× greater than community respiration, i.e., the nearshore component of the water column was net autotrophic. Benthic primary production in nearshore areas where light reached the sediments ranged between 350 and 500 mg C m⁻² d⁻¹ and was about equal to community respiration. Primary production was probably limited by the availability of dissolved inorganic N, which sometimes was below detection limits of ˜0.5 μM (mean ˜0.25 μM). Our results support the idea that autotrophic production is an important C source in neotropical rivers.