Research Highlights: Plant cover drives the activity of the microbial decomposer community and affects carbon (C) sequestration in the soil. Despite the relationship between microbial activity and C sequestration in the soil, potential inhibition of soil microbial activity by plant cover has received little attention to date. Background and Objectives: Differences in soil microbial activity between two paired stands on soil at a very early stage of formation and a common story until afforestation, can be traced back to the plant cover. We hypothesized that in a black locust (Robinia pseudoacacia L.) stand the high-quality leaf litter of the tree, and that of the blackberry (Rubus fruticosus L.) understory had an inhibitory effect on soil microbial community resulting in lower mineralization of soil organic matter compared to the paired black pine (Pinus nigra Arn.) stand. Materials and Methods: We estimated potential mineralization rates (MR), microbial (MB), and active fungal biomass (AFB) of newly-shed litter, forest floor, and mineral soil. We tested the effects of litters' water extracts on soil MR, MB, AFB and its catabolic response profile (CRP). Results: Newly-shed litter of black locust had higher MR than that of blackberry and black pine; MR, MB, and AFB were higher in forest floor and in mineral soil under black pine than under black locust. Water extracts of black locust and blackberry litter had a negative effect on the amount, activity of microorganisms, and CRP. Conclusions: The results demonstrate the potential for black locust and blackberry litter to have a marked inhibitory effect on decomposer microorganisms that, in turn, reduce organic matter mineralization with possible consequences at the ecosystem level, by increasing C sequestration in mineral soil.