Land conversion and continuous cultivation alter the belowground ecosystems, often resulting in a decrease in soil carbon (C) and biodiversity. However, to what extent those land-use changes in drylands affects soil biodiversity is still unknown. This study aims to determine effects of land-use changes on soil C and to link microbial response (focusing on 16S rRNA region and ITS) in dryland soils in Zambia. We sampled soils at three land-use changes in two sites (Lusaka, Kabwe) in Zambia. The land-use type consists of undeveloped areas (NF), agricultural land within five years of development (NC), and agricultural land for more than 10 years (OC). Lusaka soils have relatively higher clay contents (19%) than Kabwe soils (12%). Soil C and nitrogen were overall higher in Lusaka than in Kabwe. The result shows that cultivation significantly decreased both bacterial and fungal abundance in both soils. Association network analysis demonstrated that 700 bacterial ASVs and 129 fungal ASVs exhibited characteristic associations with land-use systems were detected in Lusaka soils, while only 123 bacterial ASVs and 165 fungal ASVs were obtained in Kabwe soils. Those results indicate that land-use effects on belowground biodiversity largely depend on soil inherent properties.