Afforestation of agricultural land is increasing, partly because it is an important biological method for reducing the concentration of atmospheric CO2 and potentially mitigating climate change. Rainfall patterns are changing and prolonged dry periods are predicted for many regions of the world, including southern Australia. To accurately predict land-use change potential for mitigating climate change, we need to have a better understanding of how changes in land-use (i.e. afforestation of pastures) may change the soils response to prolonged dry periods. We present results of an incubation study characterising C and N dynamics and the microbial community composition in soil collected from two tree plantings and their adjacent pastures under a baseline and reduced frequency. While the concentration of soil C was similar in pasture and tree planting soils, heterotrophic respiration was significantly lower in soil from pastures than tree plantings. Although there was little difference in the composition of the soil microbial community among any of the soils or treatments, differences in N cycling could indicate a difference in microbial activity, which may explain the differences in heterotrophic respiration between pastures and tree plantings. Soils from pastures and tree plantings responded similarly to a reduction in wetting frequency, with a decrease in microbial biomass (measured as total PLFA), and a similar reduction in heterotrophic respiration from the soil. This suggests that the responses to changes in future wetting cycles may be less dependent on land-use type than expected.