Enrichment of 13C with depth in soil organic horizons is not explained by CO2 or DOC losses during decomposition

•We analyzed the C isotope mass balance of SOC in the organic horizon.•We tested the role of DOC in enriching SOC in 13C over time.•Neither CO2 nor DOC fluxes can explain enrichment of SOC in 13C with depth.•Vertical C transport by roots or fungi may better explain the C isotope profile. Stable isotope ratios of soil organic carbon (SOC) are a potentially powerful, integrative tool for analyzing the soil C cycle. However, limited understanding of the mechanisms for C isotope fractionation in soil prevents their widespread application. Soil organic carbon (SOC) is progressively enriched in 13C with age and depth in the soil profile even though CO2 produced during soil respiration is typically enriched in 13C compared to SOC. This results in an apparent mass balance paradox. To resolve this paradox, we hypothesized that the loss of 13C-depleted DOC during soil diagenesis, the combination of microbial decomposition and abiotic processes such as leaching, can account for the increase in SOC δ13C with depth. We combined three independent approaches (field measurements, a laboratory incubation experiment, and seasonal sampling of DOC across a climate transect) to systematically evaluate the relationship between the δ13C of soil respiration, DOC, and the SOC it is derived from. However, DOC was not significantly depleted in 13C compared to SOC in any of the three approaches, and mass balance calculations indicated that the DOC flux cannot account for the full extent of 13C enrichment of SOC with depth in the soil profile. We suggest that vertical C transport by plant roots or fungi, rather than diagenesis, may be largely responsible for the observed C isotope profile. Future studies aimed at understanding these vertical transport processes should enable increased application of soil δ13C, enhancing soil biogeochemical studies.