The Uruguayan savanna ecoregion in South America is considered critically endangered mostly because of land use change and intensification to commercial agriculture and afforestation. In Uruguay, the National System of Protected Areas (SNAP) covers only ~1% of the land, while more than 90% of the land is privately owned and under different production regimes. Consequently, the implementation of conservation actions depends mostly on landowners’ willingness to collaborate and on their management capabilities. Uruguay is signatory of the CBD and has committed to achieve biodiversity conservation in a sustainable manner that does not affect local stakeholders negatively. Therefore, there is a need to develop and implement national voluntary private land conservation (VPLC) strategies to complement protected areas with other effective area-based conservation measures. VPLC has the potential to increase the area under protection, and increase ecological, economic and social connectivity, as well as the involvement of different stakeholders in landscape management. Many strategies and incentives have been developed worldwide to promote VPLC, but their success is generally context-dependent. To develop adequate VPLC policies that increase landowners’ involvement and ensure their long-term commitment it is necessary to understand their perspectives and preferences regarding biodiversity conservation. With this aim, in collaboration with the SNAP, we conducted semi-structured interviews with traditional cattle ranching landowners in a priority area for biodiversity and ecosystem services conservation in Uruguay. We followed a non-probability sampling approach consisting on stakeholders’ analysis, purposeful and snowball sampling and respondent-driven sampling. We conducted 11 interviews following the sample size saturation principle. We assessed the dependability, credibility, confirmability, and transferability of the research design, conduct, and interpretation. The results revealed landowners’ strong sense of place and their voluntary support to biodiversity conservation, given that their income and lifestyle strongly depend on the ecosystem services provided by native ecosystems (e.g. grasslands, forests). Their conservation motivations are mostly intrinsic (autonomy, competence, and relatedness; according to the self-determination theory). In order to improve their practices, they prefer facilitative incentives such as education programs and technical assistance and cost-share incentives to be able to improve their infrastructure (e.g. fencing for grassland management). Both the approach and the findings are crucial to bridge the science-implementation gap, to identify appropriate strategies to promote VPLC and to enhance landowners’ willingness to get involved in conservation actions. This was the first study to address the landowners’ perspectives on VPLC in Uruguay and the results will inform policy making at the local and national level.