We examine transformations in institutional regimes defining access to ecosystem services in Doñana, a rural region in southwestern Spain that is internationally recognized for its outstanding biocultural values. First, we review historical changes in rules, norms, and conventions defining access to ecosystem services. Second, we conduct a survey among local informants to assess the scales at which ecosystem services are supplied, demanded, and governed and discuss scale misfits in relation to historical conflicts over access to ecosystem services. We identify (1) two major periods of institutional change, characterized by conflicts between the central state and local users from enclosures of communal lands and associated restrictions in access to ecosystem services, and (2) a clash between customary governance institutions and new ones emerging with growing central state intervention and market integration. Our results suggest that multilevel governance regimes that promote coordination and institutional diversity across scales while respecting local sovereignty over ecosystem management are more likely to prevent environmental conflicts and to produce better outcomes regarding the long-term sustainable use of ecosystem services.