Several decades ago, many former pastures in Central Europe were afforested or colonized by trees after being abandoned. Knowledge of the effects of tree species on soil properties is important for planning of the composition of future forests. In this regard, a research location in Vrchdobroč (Central Slovakia), which is former agricultural land used as pasture, enables the exploration of ecosystem processes and properties in stands of different tree species after afforestation. The goal of our study was to find out whether changes in soil properties, including soil microbial activity and diversity among different stands, were already observable 45 years after the afforestation, and how the effects differed among stands of different tree species. The study was conducted in the pure stands of Norway spruce (Picea abies L. Karst.), Douglas fir (Pseudotsuga menziesi (Mirb.) Franco), European beech (Fagus sylvatica L.) and sycamore maple (Acer pseudoplatanus L.). Multivariate analyses of physico-chemical soil properties indicated an overlap between the soils under the Douglas fir and the spruce, but a clear separation of beech from sycamore. In general, both microbial activity and diversity were, surprisingly, highest under the Douglas fir, followed by the sycamore, with the beech and the spruce showing mostly lower values.