Persistent biogeochemical signals of land use-driven, deep root losses illuminated by C and O isotopes of soil CO2 and O2

Replacing long-lived, rarely disturbed vegetation with short-lived, frequently disturbed vegetation is a widespread phenomenon in the Anthropocene that can influence ecosystem functioning and soil development by reducing the abundance of deep roots. We explore how sources and fate of soil CO₂ vary with organic substrate source, abundance of respiring biota (i.e., roots and soil microbes), season, and soil depth. We quantified multiple isotopic signatures of CO₂ (δ¹³C, Δ¹⁴C, δ¹⁸O) as well as concentrations and δ¹⁸O of free O₂ in the upper 5 m of soil at sites where root abundances and soil organic C have been previously quantified: in late-successional forests, cultivated fields, and ~ 80 y old regenerating pine forests growing on previously cultivated land. We hypothesized that soil CO₂sources would vary across soil depth and land cover, reflecting varying abundances of organic substrates, and seasonally as the dominance of root vs. microbial CO₂ production changes through the year. δ¹³C–CO₂ revealed respiration of C4-derived substrates in cultivated fields particularly during the growing season. This effect was not evident in soils of regenerating pine or older hardwood forests, suggesting that ~ 80 y of pine inputs to reforested soils have been sufficient to dominate microbial substrate selection over any remnant, historic agricultural C4 inputs. Δ¹⁴C–CO₂ diverged by land use at 3 and 5 m, indicating that more recently-produced photosynthate is available for mineralization in forests compared to cultivated plots, and in late-successional forests compared to regenerating pine forests. At 1.5, 3, and 5 m in forested plots we observed evidence of respiratory demands on soil pore space O₂. In these soils, we observed declines in [O₂] compared to other depths and to the agricultural plots and concurrent increases in δ¹⁸O of free O₂, consistent with the idea that roots and heterotrophic soil microbes are more active where photosynthate is more available. The δ¹⁸O–CO₂ values, a likely proxy for δ¹⁸O of soil porewater, exhibited ¹⁸O enrichment during the winter, when many sampling wells were flooded, compared to growing season values. These data suggest an isotopically-distinct and laterally-flowing source of CO₂-laden porewater during winter months. Combined, these datasets document how ~ 80 y of forest regeneration can provide sufficient C inputs to mask any microbial mineralization of decades-old organic inputs, but belowground C inputs still lag those of late successional forests. We also infer that lateral and vertical flows of water can serve as a sink for biotically-generated CO₂, and that where deep soil [CO₂] is lower due to lower root and microbial activities, production of carbonic acid is also diminished. Where reaction rates are weathering limited, a paucity of deep roots imposed by anthropogenic land cover change thus may limit the production of this agent of soil development and the C sink represented by the silicate weathering it can promote. The data suggest deep and persistent effects of the loss of deeply rooted long-lived vegetation on deep soil C storage and transformations that promote acid-dissolution weathering reactions that help form soil itself.