Most of traditional knowledge about plants and their uses is fast disappearing because of socio-economic and land use changes. This trend is also occurring in bio-cultural refugia, such as mountain areas. New data on Traditional Ethnobotanical Knowledge (TEK) of Italian alpine regions were collected relating to three valleys (Cogne, Valsavarenche, Rhêmes) of the Gran Paradiso National Park. Extensive dialogues and semi-structured interviews with 68 native informants (30 men, 38 women; mean age 70) were carried out between 2017 and 2019. A total of 3918 reports were collected, concerning 217 taxa (including 10 mushrooms, 1 lichen) mainly used for medicinal (42%) and food (33%) purposes. Minor uses were related to liquor making (7%), domestic (7%), veterinary (5%), forage (4%), cosmetic (1%) and other (2%). Medicinal plants were used to treat 14 ailment categories, of which the most important were respiratory (22%), digestive (19%), skin (13%), musculoskeletal (10%) and genitourinary (10%) diseases. Data were also evaluated by quantitative ethnobotanical indexes. The results show a rich and alive traditional knowledge concerning plants uses in the Gran Paradiso National Park. Plants resources may provide new opportunities from the scientific point of view, for the valorization of local products for health community and for sustainable land management.