Annual ryegrass (Lolium multiflorum Lam.) and crimson clover (Trifolium incarnatum L.) were pulse-labeled with ¹³C-CO₂ in the field between the initiation of late winter growth (mid-February) and through flowering and seed formation (late May). Straw was harvested after seed maturation (July), and soil containing ¹³C-labeled roots and root-derived C was left in the field until September. ¹³C-enriched and ¹³C-unenriched straw residues of each species were mixed in factorial combinations with soil containing either ¹³C-enriched or ¹³C-unenriched root-derived C and incubated in the field for 10 months. The contributions of C derived from straw, roots, and soil were measured in soil microbial biomass C, respired C, and soil C on five occasions after residue incorporation (September, October, November, April, and June). At straw incorporation (September), 25–30% of soil microbial biomass C was derived from root C in both ryegrass and clover treatments, and this value was sustained in the ryegrass treatment from September to April but declined in the clover treatment. By October, between 20 and 30% of soil microbial biomass C was derived from straw, with the percentage contribution from clover straw generally exceeding that from ryegrass straw throughout the incubation. By June, ryegrass root-derived C contributed 5.5% of the soil C pool, which was significantly greater than the contributions from any of the three other residue types (about 1.5%). This work has provided a framework for more studies of finer scale that should focus on the interactions between residue quality, soil organic matter C, and specific members of the soil microbial community.