The decomposition of soil organic carbon (SOC) is intrinsically sensitive to temperature. However, the degree to which the temperature sensitivity of SOC decomposition (as often measured in Q 10 value) varies with soil depth and labile substrate availability remain unclear. This study explores (1) how the Q 10 of SOC decomposition changes with increasing soil depth, and (2) how increasing labile substrate availability affects the Q 10 at different soil depths. We measured soil CO 2 production at four temperatures (6, 14, 22 and 30°C) using an infrared CO 2 analyzer.Treatments included four soil depths (0-20, 20-40, 40-60 and 60-80 cm), four sites (farm, redwood forest, ungrazed and grazed grassland), and two levels of labile substrate availability (ambient and saturated by adding glucose solution). We found that Q 10 values at ambient substrate availability decreased with increasing soil depth, from 2.0-2.4 in 0-20 cm to 1.5-1.8 in 60-80 cm. Moreover, saturated labile substrate availability led to higher Q 10 in most soil layers, and the increase in Q 10 due to labile substrate addition was larger in subsurface soils (20-80 cm) than in surface soils (0-20 cm). Further analysis showed that microbial biomass carbon (MBC) and SOC best explained the variation in Q 10 at ambient substrate availability across ecosystems and depths (R 2 = 0.37, P \ 0.001), and MBC best explained the variation in the change of Q 10 between control and glucose addition treatment (R 2 = 0.14, P = 0.003).Xueyong Pang and Biao Zhu contributed equally to this work.