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We measured the natural abundances of radiocarbon (Δ14C) in macroinvertebrates, fishes, and their potential food sources, collected from the upper and lower reaches of six temperate streams in Lake Biwa basin (central Japan), three of which flow on limestone bedrock. Several carbon storage reservoirs in the watersheds show distinctive Δ14C signatures (e.g., ancient carbonate rocks, -1000‰; modern atmospheric CO2, +50‰). Our analyses showed that the Δ14C values for periphytic algae range from -361‰ to +21‰, reflecting14C-depleted signals from watershed storage reservoirs (carbonate rocks and/or soils). In contrast, the Δ14C values for coarse particulate organic matter (CPOM) range from -6‰ to +62‰, reflecting 14C-enriched signals from modern atmospheric CO2. The periphyton from streams on limestone bedrock was more 14C-depleted than that from streams in non-limestone areas, although the Δ14C values for periphyton from the latter were less than modern atmospheric 14CO2 concentration. The Δ14C values for most of the consumers were between those for periphyton and CPOM. Based on a Δ14C two-source mixing models, the results suggested that the grazers rely on periphyton, while the carbon source for collectors and predators shifts from CPOM in the upper reaches of streams to periphyton in the lower reaches. The Δ14C signature can trace carbon from watershed storage reservoirs to benthic production, which suggests that stream food webs are composed of mixtures of carbon originating from various sources of different ages. [ABSTRACT FROM AUTHOR]