The increasing interest in bio-districts is part of the debate on the capacity to integrate agri-food systems and territory in order to improve the quality of life in rural communities. Considering the goals of developing and promoting an innovative territorial rural development approach, the bio-district can become a process toward a more sustainable model represented by the agroecological agriculture system. The paper presents a case study of the Parma bio-district through the approach of a Localized Agri Food System (LAFS) to verify whether bio-districts can be a tool for scaling up towards agroecology. Stakeholder classification and analysis are conducted using an influence–interest matrix. We identified four groups of stakeholders in relation to their interests and power to influence the process. In the case of the Parma bio-district the role of local institutions in dialogue with consumers and producers’ associations is crucial for success. We conclude that bio-districts can be a tool for a scaling-up towards agroecology since they can facilitate a synergetic relation between organic and agroecological agriculture, spreading organic agriculture more widely around the local area. However, the involvement of a wide variety of different stakeholders means that governance is a key element in facilitating “cross fertilization” and preventing the process from becoming purely formulaic.