Sustainable feedstock supply is a critical issue for the bioenergy sector. One concern is that feedstock production will impact biodiversity. We analyze how this concern is addressed in assessments of biomass supply potentials and in selected governance systems in the EU and Brazil, including the EU Renewable Energy Directive (RED), the EU Common Agricultural Policy (CAP), and the Brazilian Forest Act. The analysis focuses on grasslands and includes estimates of the amount of grassland area (and corresponding biomass production volume) that would be excluded from cultivation in specific biodiversity protection scenarios. The reviewed assessments used a variety of approaches to identify and exclude biodiverse grasslands as unavailable for bioenergy. Because exclusion was integrated with other nature protection considerations, quantification of excluded grassland areas was often not possible. The RED complements and strengthens the CAP in terms of biodiversity protection. Following the RED, an estimated 39%–48% (about 9–11 Mha) and 15%–54% (about 10–38 Mha) of natural and non‐natural grassland, respectively, may be considered highly biodiverse in EU‐28. The estimated biomass production potential on these areas corresponds to some 1–3 and 1.5–10 EJ/year for natural and non‐natural grassland, respectively (depending on area availability and management intensity). However, the RED lacks clear definitions and guidance, creating uncertainty about its influence on grassland availability for bioenergy feedstock production. For Brazil, an estimated 16%–77% (about 16–76 Mha) and 1%–32% (about 7–24 Mha) of natural and non‐natural grassland, respectively, may be considered highly biodiverse. In Brazil, ecological–economic zoning was found potentially important for grassland protection. Further clarification of grassland definitions and delineation in regulations will facilitate a better understanding of the prospects for bioenergy feedstock production on grasslands, and the impacts of bioenergy deployment on biodiversity.