Aggregate formation and soil carbon sequestration by earthworms at the ORNL FACE experiment

Earthworms have an important role in soil carbon sequestration, but their contribution to carbon sequestration in soils exposed to elevated atmospheric CO₂ concentrations has been largely overlooked. Previous studies at the Oak Ridge National Laboratory Free Air CO₂ Experiment (ORNL FACE) site showed that the formation of soil aggregates is a key mechanism for soil carbon sequestration. We did a microcosm experiment to quantify earthworm-mediated aggregate formation and compare between two earthworm species with different feeding habits (endogeic vs. epi-edogeic). In addition, we wanted to identify the carbon source (soil, leaf litter or root litter) within aggregates formed by earthworms. We used ¹³C-depleted soil and ¹⁵N-enriched sweetgum (Liquidambar styraciflua) leaf and root litter collected from the ORNL FACE site to assess soil aggregate formation of the native, endogeic earthworm Diplocardia sp. and European, epi-endogeic earthworm Lumbricus rubellus. Both earthworm species are present at the ORNL FACE site. We crushed, sieved (< 250 μm) soil and prepared four treatments: (I) soil only; (II) soil and plant material; (III) soil, plant material and Diplocardia sp.; (IV) soil, plant material and L. rubellus. All treatments were at 30% water content and temperature was maintained at 20°C. The incubation period lasted 26 days. We measured aggregate size distribution, total aggregate carbon content and ¹³C and ¹⁵N to elucidate aggregate carbon source. Newly formed soil macroaggregates (> 250 μm) were higher in treatments with earthworms (III and IV) than in treatments without earthworms (I and II) (p = 0.02). Within macroaggregates, most of the carbon was soil-derived. Leaf and root-derived carbon was found in treatment IV only. Our results suggest that earthworms at the ORNL FACE site directly contribute to the formation of soil aggregates, thus contributing to soil carbon sequestration. Carbon source within macroaggregates correspond with earthworm feeding habits, with endogeic earthworms (Diplocardia sp.) feeding mostly on mineral soil and epi-endogeic earthworm (L. rubellus) feeding on both plant residues and soil organic matter.