Carbon sequestration in agricultural soils may help to reduce global greenhouse gas concentrations, but building up soil carbon levels requires accumulating organic matter faster than it is lost via heterotrophic respiration. Using field and laboratory studies, this study sought to elucidate how tillage, the below-ground incorporation of cover crop residue, and soil macroporosity affect soil respiration and residue decomposition rates. In the field, residue from a cover crop mixture of barley (Hordeum vulgare) and crimson clover (Trifolium incarnatum) was placed into litter bags that were left on the surface versus incorporated into the soil at three depths (4, 8 or 12 cm), while the laboratory study compared surfaceplaced versus incorporated litter (8 cm depth). To assess tillage effects on cover crop decomposition, the field study simulated no-till and conventional tillage treatments, while the laboratory and field studies both included treatments in which artificial soil macropores were created. The field study showed that conventional tillage and the presence of macropores enhanced soil respiration, while in the laboratory study, incorporating cover crop residue resulted in higher soil respiration and faster litter decomposition rates. Additionally, the laboratory measurements showed that macropores increased soil respiration in wet conditions, likely by enhancing oxygen diffusion. Thus, organic matter incorporation and macropores may represent important factors that affect soil respiration and carbon dynamics.