Erythrina velutina Willd. (Fabaceae: Papillionoideae) is a pioneer species found in tropical and subtropical regions of the world that has medicinal properties and that is used in reforestation projects. This species is rare in some areas of northeastern Brazil. This study aimed to characterize and compare genetic structures of natural and restored populations of E. velutina, with a focus on the selection of tree seeds. A total of 108 individuals from five natural populations and one restored population were analyzed using ISSR markers, resulting in 407 polymorphic fragments. A high rate of polymorphism was observed in the restored population. The highest genetic variability was identified within populations (82%). Genetic bottleneck tests were significant for the Carmópolis/Rosário do Catete and Laranjeiras natural populations along with the Laranjeiras restored population. Genetic distances significantly correlated with spatial distance. Only the restored population retained unique alleles. Similarly, increased genetic distance was observed in individuals of the restored populations compared to the other populations. Observed genetic variation in both natural and restored populations of E. velutina was moderate, thus enabling selection of divergent trees from those trees supplying seeds. Environmental protection and management of these areas is necessary for the maintenance of these individuals and subsequent reproduction. We recommend suggestions for E. velutina conservation, since the restoration model adopted in this study did not promote the development of the specimens until the reproductive stage in a fashion that aims to augment the soil seed bank supply, as is suggested for pioneer species.Ribeirão Preto, SP