The Trentino-South Tyrol region is a special statute region of northeastern Italy. This territory is of particular interest for its morphology, flourishing vegetation, and history, having been a meeting area among different civilizations. Hence, Trentino is characterized by an ethnic plurality and a rich ethnobotanical knowledge, even if the available information is fragmentary, widely dispersed, and often guarded in oral popular culture. To fill this gap, in the present work 200 subjects were interviewed using an ethnobotanical survey. The resulting 817 citations referred to 64 native species, used either for human or animal health or for domestic purposes. As a second step, for each plant exploited for medicinal purposes, local importance was evaluated by calculating their relative frequency of citation. Moreover, the main traditional preparations were discussed. Among them, the most cited and exploited ones are Achillea millefolium, Arnica montana, Hypericum perforatum, Malva sylvestris, Pinus mugo, and Satureja montana, for which a deeper analysis has been performed. Lastly, the ethnobotanical knowledge of the plants growing in this territory will add a piece to the mosaic of traditional medicine in Italy and may lay the foundation for a nature-aided drug discovery process.