ContextHuman-induced changes in landscape structure are among the main causes of biodiversity loss. However, despite their important contribution to biodiversity and ecosystem functioning, microbes – and particularly protists – remain spatially understudied. Furthermore, soil microbiota are most often related to the local soil properties, whereas the influence of the surrounding landscape is rarely assessed, calling for microbial landscape ecology studies.ObjectivesHere, we assessed for the first time the effect of changes in landscape composition and configuration on soil protist diversity in meadows in the Swiss western Alps.MethodsWe sampled 178 plots along an elevation gradient representing a broad range of climatic, edaphic, and land use conditions, and measured landscape structures in several neighbourhood windows of increasing size.ResultsOur results showed that landscape structures between 500 m and 2000 m around plot centres better predicted microbial diversity than local edaphic and climatic descriptors, with the percentage cover of open habitat and meadows and, to a lesser extent, edge density being the most important descriptors. The patch level descriptors explained the least amount of variation in protist diversity, what is in contradiction with macro-organims where patch size and shape strongly impact the population dynamics. This study also highlights different responses of protist functional groups to changes in landscape structure.ConclusionsOur results importantly show the significance of landscape structure for microbial diversity and the potentially negative consequences of land use change on soil microbial communities and their associated functions.