Partitioning the soil surface CO2 flux (RS) flux is an important step in understanding ecosystem-level carbon cycling, given that RS is poorly constrained and its source components may have different sensitivities to climate change. Trenched plots are an inexpensive but labor-intensive method of separating the RS flux into its root (autotrophic) and soil (heterotrophic) components. This study tested if various methods of plant suppression in trenched plots affected RS fluxes, quantified the RS response to soil temperature and moisture changes, and estimated the heterotrophic contribution to RS. It was performed in a boreal black spruce (Picea mariana) plantation, using a randomized complete block design, during the 2007 and 2008 growing seasons. Trenched plots had significantly lower RS than control plots, with differences appearing ∼100 days after trenching; spatial variability doubled immediately after trenching but then declined throughout the experiment. Most trenching treatments had significantly lower (by ∼0.5 μmol CO2 m−2 s−1)RS than the controls, and there was no significant difference in RS among the various trenching treatments. Soil temperature at 2 cm explained more RS variability than did 10-cm temperature or soil moisture. Temperature sensitivity (Q10) declined in the control plots from ∼2.6 (at 5 °C) to ∼1.6 (at 15 °C); trenched plots values were higher, from 3.1 at 5 °C to 1.9 at 15 °C. We estimated RS for the study period to be 241 ± 40 g C m−2, with live roots contributing 64% of RS after accounting for fine root decay, and 293 g C m−2 for the entire year. These findings suggest that laborious hand weeding of trenched plot vegetation may be replaced by other methods, facilitating future studies of this large and poorly-understood carbon flux.