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Soil microbial communities directly affect soil functionality through their roles in the cycling of soil nutrients and carbon storage. Microbial communities vary substantially in space and time, between soil types and under different land management. The mechanisms that control the spatial distributions of soil microbes are largely unknown as we have not been able to adequately upscale a detailed analysis of the microbiome in a few grams of soil to that of a catchment, region or continent. Here we reveal that soil microbes along a 1000 km transect have unique spatial structures that are governed mainly by soil properties. The soil microbial community assessed using Phospholipid Fatty Acids showed a strong gradient along the latitude gradient across New South Wales, Australia. We found that soil properties contributed the most to the microbial distribution, while other environmental factors (e.g., temperature, elevation) showed lesser impact. Agricultural activities reduced the variation of the microbial communities, however, its influence was local and much less than the overall influence of soil properties. The ability to predict the soil and environmental factors that control microbial distribution will allow us to predict how future soil and environmental change will affect the spatial distribution of microbes.