Abstract: Disentangling the autotrophic and heterotrophic components of soil CO2 efflux is critical to understanding the role of soil system in terrestrial carbon (C) cycling. In this study, we combined a stable C-isotope natural abundance approach with the trenched plot method to determine if root exclusion significantly affected the isotopic composition (delta C-13) of soil CO2 efflux (R-S). This study was performed in different forest ecosystems: a tropical rainforest and two temperate broadleaved forests, where trenched plots had previously been installed. At each site, R-S and its delta C-13 (delta C-13(Rs)) tended to be lower in trenched plots than in control plots. Contrary to R-S, delta C-13(Rs) differences were not significant. This observation is consistent with the small differences in delta C-13 measured on organic matter from root, litter and soil. The lack of an effect on delta C-13(Rs) by root exclusion could be from the small difference in delta C-13 between autotrophic and heterotrophic soil respirations, but further investigations are needed because of potential artefacts associated with the root exclusion technique.