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The objective of this work was to evaluate the genetic variability distribution of umbu tree (Spondias tuberosa), within Brazilian Semi-Arid region, based on AFLP markers, in order to suggest prospecting and preservation strategies for this species. Sixty-eight umbu trees of 15 ecogeographic regions were analyzed for 141 polymorphic AFLP bands, through the UPGMA dendrogram and the multidimensional scaling (MDS), based on Jaccard’s coefficient. Analysis of molecular variance was accomplished by total decomposition among and within ecogeographic regions. The dendrogram presented co-phenetic value of 0.96, while the MDS presented 0.25 for the badness-of-fit. Umbu genetic variability was estimated in 0.3138, indicating a large variance among individual groups. Specific clusters were observed in six ecogeographic regions, and some individual pairs were observed in the other regions, with no specific clustering by sampling place, which indicates that the genetic variability of umbu tree is not uniformly distributed within Brazilian Semi-Arid. Strategies are suggested to set a larger number of protection areas for in situ conservation, or a smaller number of umbu individuals sampled, in various ecoregion units, for ex situ genetic variability conservation of this species.