The identification of research questions with high relevance for biodiversity conservation is an important step towards designing more effective policies and management actions, and to better allocate funding among alternative conservation options. However, the identification of priority questions may be influenced by regional differences in biodiversity threats and social contexts, and to variations in the perceptions and interests of different stakeholders. Here we describe the results of a prioritization exercise involving six types of stakeholders from the Mediterranean biome, which includes several biodiversity hotspots spread across five regions of the planet (Europe, Africa, North and South America, and Australia).