Better balancing agricultural production and biodiversity conservation is a central goal for many landscapes. Yet, empirical work on how to best achieve such a balance has focused mainly on the local scale, thereby disregarding that landscape context might mediate biodiversity-agriculture trade-offs. Focusing on vertebrates in the Argentine Chaco, we evaluate how trade-offs between agriculture and biodiversity vary with landscape context, from landscapes where agricultural and natural areas are separated to landscapes where both are interspersed. We modelled the distributions of 226 vertebrates and use the resulting maps to describe the species richness of ecosystem-service providing guilds. We calculated three agricultural intensity metrics, and evaluated how both species richness and agricultural intensity vary along a gradient of landscape configuration, while controlling for landscape composition. Species richness and agricultural yields both varied with landscape configuration. Biodiversity was highest in mixed landscapes where agricultural and natural area are interspersed, whereas agricultural yields showed a more heterogeneous response, with some yield metrics highest in mixed and others in separated landscapes. As a result, agriculture/biodiversity trade-offs depended strongly on landscape configuration, irrespective of landscape composition. We also identified large areas with low vertebrate richness and agricultural yields, suggesting considerable potential for improving in at least one dimension. Agriculture/biodiversity trade-offs varied with landscape configuration, suggesting that landscape design can balance these trade-offs. Our simple and broadly applicable approach can provide baseline information for landscape planning aimed at realizing co-benefits between agriculture and biodiversity—in the Gran Chaco and elsewhere.