ABSTRACT.- Mixed systems of grain and livestock production based on pasture-crop rotations are a promising strategy to promote agriculture resilience and allow an ecological intensification of agriculture yet little is known about underlying processes in soil. To test the hypothesis that pasture-crop rotations preserve soil structure and select for beneficial soil and rhizosphere microbiota, supporting soil health and grain production, a long-term field experiment under no-tillage was studied. The experiment evaluated a gradient of land use intensities and vegetation diversities, from highly intensive continuous cropping to the least intensive system i.e. a nearby natural grassland, with two intermediate land use intensities i.e. short pasture-crop rotation and long pasture-crop rotation. Soil health was assessed based on soil physicochemical properties, microbial (Bacteria/Archaea and Fungi) community diversity and composition and oat performance. Pasture-crop rotations preserved soil bulk density and larger aggregates better than continuous cropping. High-throughput amplicon sequencing of 16S rRNA gene and ITS fragments revealed that the pasture-crop rotations fostered taxa that are associated with soil structure maintenance and selected potential plant-beneficial bacterial genera in the oat rhizosphere (i.e. Bosea, Devosia and Microbacterium), that may have contributed to the observed increase in N uptake, N accumulation and biomass in oat. In summary, this study shows that pasture-crop rotations are an ecologically sustainable alternative to continuous cropping in the Uruguayan Pampa biome. © 2024 The Authors