Currently, with the rise in agricultural village tourism, tour navigation aided by mobile devices has become a new trend. This research aims to develop mobile tourism navigation suitable for agricultural village tourism and enhance the travel experience of tourists wherein food and agriculture education, along with relevant ecological knowledge, could be learned. An understanding of system usability and workload was gained through field experiments and semi-structured interviews involving fifteen youth-aged users, fifteen active aging elderly users, and operational feeling. The researchers also integrated comments from tourism service operators. A design for the mobile ecological navigation for agricultural village tourism was proposed and a passive and game-exploring ecological mobile navigation was then developed. The number of operational steps for passive ecological navigation was kept low to relieve the physical load, especially for middle-aged users. The navigation is suitable for ordinary tourists and easily directs them to ecological fields. However, game-exploring mobile ecological navigation features more operational steps. Although the active aging elderly group faced a higher physical load, after they became familiar with the steps, they could get into situations without much effort or difficulty. Furthermore, the navigation processes were designed with highly interesting elements to enhance the use experience for the youth-aged group and trigger the exploring motivation of tourists. The design is also suitable for ecological education activities involving teachers and students. Mobile ecological navigation can not only improve the quality of navigation service, but also deliver to tourists a tour experience totally different from what they have experienced.