Crop wild relatives (CWR) are valuable gene pools for crop improvement and offer unique potential and opportunity for enhancing food security and adaptation to climate change. However, current actions towards conservation of plant genetic resources in Zambia do not adequately cover CWR occurring in the country. The article describes the process leading to the development of a national strategic action plan (NSAP) for the conservation and sustainable use of priority CWR in Zambia. Based on 59 prioritized crops, a partial checklist of 459 CWR taxa was generated from the national flora checklist of 6305 taxa. The generated CWR taxa were prioritized based on the socio-economic value of the related crop, their utilization potential in crop improvement, relative distribution and threat status to produce 30 prioritized CWR taxa. Occurrence data were compiled for all CWR inventory taxa and used in spatial analyses to establish species distribution, species richness, gaps in in situ conservation and genebank collections, and to identify priority sites for in situ conservation and ex situ collecting. Consistent with the national developmental agenda, along with the contribution of national stakeholders, spatial analyses of occurrence data of priority CWR taxa are valuable input for the development of the NSAP for the conservation and sustainable use of the priority CWR.