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As one of the most dominant ecosystems of urban green space, turfgrasses provide a wide range of ecosystem services. However, little is known about the interactions of microbial communities in turfgrass soils and how these interactions respond to expanding development of impervious surfaces during watershed urbanization. In this study, we analyzed bacterial communities and their co-occurrence patterns in turfgrass soils along an urbanization gradient as measured by the proportion of impervious surfaces in Jiulong River watershed in Fujian, China. Results show that the diversity and network size of bacterial communities negatively associated with impervious surfaces. The bacterial communities showed non-random co-occurrence patterns, with more intra-module connections observed for urbanized networks. The co-occurrence network with distinct modules of soil samples with contrasting land cover imperviousness suggested different functional organizations with altered microbial nitrogen processes. Structural equation modelling revealed that watershed impervious surfaces had indirect impacts on microbial connectivity by altering soil properties, including pH, temperature, moisture, C/N and nitrate (NO 3 −). Moreover, impervious surfaces affected microbial connectivity far more than human population density. Our study highlights the significance of human disturbances in affecting microbial interactions and assemblies in turfgrass ecosystems through impervious surfaces and provides benefits for sustainable urban planning and management at a watershed scale. Image 1 • Increased impervious surfaces reduce microbial diversity and network size. • Bacterial communities showed more intra-module connections in urbanized networks. • Impervious surfaces affect micro-connectivity far more than population density. • Changes in micro-connectivity with land cover urbanization were driven by soil pH. Land cover urbanization significantly impact microbial connectivity in turfgrass soils. [ABSTRACT FROM AUTHOR]