Relatively few studies have documented the impacts of afforestation, particularly production forestry, on belowground carbon dioxide (CO₂) effluxes to the atmosphere. We evaluated the changes in the soil CO₂ efflux—a proxy for soil respiration (Rs)—for three years following a native grassland conversion to eucalypt plantations in southern Brazil where minimum tillage during site preparation created two distinct soil zones, within planting row (W) and between-row (B). We used root-exclusion and carbon (C)- isotopic approaches to distinguish Rs components (heterotrophic-Rh and autotrophic-Ra respirations), and a CO₂ profile tube (1-m deep) to determine the concentration (CO₂]) and isotopic C signature of soil CO₂ (δ¹³CO₂]). The soil CO₂ efflux in the afforested site averaged 0.37 g CO₂ m^?2 h^?1, which was 56% lower than the soil CO₂ efflux in the grassland. The δ¹³CO₂ in the afforested site ranged from ? 14.1‰ to ? 29.4‰, indicating a greater contribution of eucalypt-derived respiration (both Rh and Ra) over time. Higher soil CO₂ efflux and lower CO₂] were observed in W than B, indicating that soil preparation creates two distinct soil functional zones with respect to C cycling. The CO₂] and δ¹³CO₂] decreased in both zonal positions with eucalypt stand development. Although the equilibrium in C fluxes and pools across multiple rotations is needed to fully account for the feedback of eucalypt planted forests to climate change, we provide quantitative information on soil CO₂ dynamics after afforestation and show how soil preparation can leverage the feedback of planted forests to climate change.