Stable carbon isotope ratios of total organic matter were measured in two cores collected from the Kashiru peatbog in Burundi, Equatorial Africa. The record, which spans at least the last 40,000 yr, documents the C3-C4 biomass balance in the organic sediment. Among the major modern peat formers, most plants are C3 species and are characterized by δ13C values of -25.5 ± 2.3% (vs PDB). The C4 plants, which are characterized by higher δ13C values (-11.3 ± 0.7%) belong to the Gramineae (Miscanthidium sp.) and Cyperaceae families (Cyperus latifolius, C. papyrus, Pycreus nigricans). In the fossil record, δ13C values of total organic matter vary between -28 and -15% in response to the relative fluxes of C3 and C4 plants. Before 30,000 yr B.P., low δ13C values (-23.5 ± 1.1%) match high arboreal pollen contents. From 30,000 to 15,000 yr B.P., higher δ13C values (-17.6 ± 1.1%) correspond to a significant increase in percentages of grass pollen. During this episode, a short and sharp shift toward lighter carbon isotopic compositions at 21,000 yr B.P. is synchronous with higher input of arboreal pollen. From 15,000 to 12,000 yr B.P., the 13C content decreases (δ13C = -22.9 ± 1.4%). This shift, which cannot be explained by an increase in the arboreal vegetation, could be explained by the spreading of C3 Gramineae or C3 Cyperaceae. The interval from 12,000 to 7000 yr B.P. is poorly documented in these cores due to much lower organic matter accumulation. Low δ13C values (δ13C = -25.2 ± 1.3%) are observed from 7000 to 5000 yr B.P., when the pollen data show development of C3 mountain forest. The Late Holocene is characterized by a mixed C3-C4 organic matter accumulation (δ13C = -20.9 ± 1.6%). This study depicts a change in the dominant photosynthetic pathway among the herbaceous components, notably at the glacial-interglacial transition, when C3 plants were favored by increased water supply and/or higher atmospheric CO2 concentration.