Identifying which ecosystem services are relevant to different stakeholders and understanding stakeholders' perceptions of such services is useful for making informed decisions, especially in regions of the world where the achievement of biodiversity conservation goals is threatened by economically productive activities. In this article, we assess social preferences for ecosystem services in a biodiversity hotspot in central Chile. We use a consultative case study to ask local stakeholders (n = 70) from the Campana Penuelas Biosphere Reserve to identify the most important ecosystem services the area provides for them and inquire about the perceived vulnerability of the services to changes in the future. We also explore the association between the perceived importance of ecosystem services and the sociodemographic and cultural characteristics of the respondents, which allows us to identify contrasting stakeholder perceptions of different ecosystem services. The most important services for local actors were the drinking water, fresh air and climate change control, genetic pool of plant communities in central Chile, and educational value. From the perspective of local actors, the services that could be threatened by negative changes in the future in terms of their provision included the possibilities of developing conservation activities focused on iconic threatened animal and plant species, water regulation, food from agriculture, and drinking water. Contrasting perceptions about the importance of ecosystem services emerged among stakeholders. While small farmers and members of local organizations attributed higher importance values to provisioning services, scientists and rangers and administrators of protected areas as well as teachers, NGO members and local government employees attributed more importance to the regulating and cultural services associated with threatened species. Our results can serve as a source of information for the planning and decision-making processes related to the search for socially and ecologically sustainable solutions for land use management.