Abstract: Riding Mountain Biosphere Reserve (RMBR) was founded in 1986, 53 years after the establishment of Riding Mountain National Park (RMNP). Currently the RMBR is one of 18 biosphere reserves in Canada, three of which are located in the Prairie region. The reserve has a total area of approximately 13,310 km2 comprising the RMNP core area measuring 2,700 km2, a buffer zone of 268 km2, and a transition zone or cooperation area of 10,342 km2. Fourteen municipalities and four First Nation reserves are located within the transition zone. Three of the reserves (Rolling River, Keeseekoowenin, and Waywayseecappo) are located in the southern part of the RMBR. This paper focuses on the complex ecological, demographic, cultural, socio-economic, and political-administrative framework and relationships between RMNP, the municipalities, and the First Nation reserves. ‘Baskets’ of complementarities, mutual opportunities and benefits contrast with multiple challenges, diverging interests, and potential sources of conflict. Recently, enhanced communication, closer interactions, and joint projects have characterized relationships, at least in some areas between the National Park, and some of the municipalities and First Nation reserves. However, genuine environmental commitment and protection, as well as a strong, diversified economic base, and a harmonious coordination of these principal objectives may require an even stronger effort and cooperation between all of the regional stakeholders.