The aim of this study was for university consultants to apply and critically evaluate Appreciative Inquiry (AI) as a methodological approach to collaboration with the national organisation for promoting sport and physical activity in Wales, a Welsh national governing body for sport, and a Welsh local authority department responsible for sport and leisure, to develop a physical literacy (PL) programme for primary schools. Three phases of an AI cycle; the "discovery" phase, the "dream" phase, and the "design" phase were the focus of this study, within which the consultants acted as 'boundary spanners', to promote collaboration between the different organisations, over a period of six months. In the "discovery" and "dream" phases data was gathered through AI conversations, written accounts and observation, whilst a collaborative meeting and specific follow up actions were employed for the "design" phase. Qualitative thematic analysis revealed key messages for the use of AI in such collaborative projects, and some interesting insight into the complexities of this methodological approach. Overall, in considering the key contribution to knowledge of this study, AI was shown to be an effective approach to adopt in multi-organisational collaborative ventures of this nature. However, the collaborative structures were often intricate and dynamic in nature, resulting in some challenges for the consultants convening them. The practical implications for those wishing to conduct AI, are that such approaches require large amounts of resources in the form of energy, commitment, skill and nurturing on the part of the leaders.