This paper examines the characteristics of the process leading to collaboration among region management organizations (RMOs) and the cooperative strategies related to business operations in the Risyu region of Ena City, Gifu Prefecture. It also explores the conditions necessary for agricultural support through inter-RMO collaboration. While each district in the Risyu region had independently managed its regional affairs, a shared sense of urgency about the region’s future prompted the formation of inter-RMO partnerships. This collaboration was achieved through the development of a joint action plan and its approval by local communities. In the realm of business, a strategy that prioritized cost-saving cooperative projects and considered potential local government support facilitated early implementation, effectively demonstrating the benefits of collaboration. However, significant disparities in the agricultural management conditions of the three districts hindered the realization of unified agricultural support through inter-RMO cooperation.