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Anadenanthera (Fabaceae) is endemic to the Neotropics and consists of two tree species: A. colubrina (Vell.) Brenan and A. peregrina (L.) Speg. This study examined the mating system and contemporary gene flow of A. colubrina (Acol) and A. peregrina (Aper) in a highly fragmented area of the Atlantic Forest to provide valuable information that informs conservation strategies. Reproductive adults from forest remnants [nA. colubrina = 30 (2.7 ha), nA. peregrina = 55 (4.0 ha)] and progeny-arrays (nA. colubrina = 322, nA. peregrina = 300) were genotyped for seven nuclear microsatellite markers. Mating system analyses revealed that A. colubrina is a mixed mating species (tm = 0.619) while A. peregrina is a predominantly outcrossing species (tm = 0.905). For both Anadenanthera species, high indices of biparental inbreeding were observed (Acol = 0.159, Aper = 0.216), resulting in low effective pollination neighborhood sizes. Categorical paternity analysis revealed different scales of pollen dispersal distance: the majority of crossings occurring locally (i.e., between nearby trees within the same population), with moderate pollen dispersal coming from outside the forest fragments boundaries (Acolmp = 30%, Apermp = 35%). Nevertheless, pollen immigration from trees outside the populations for both species suggests that the populations are not reproductively isolated. This study highlights the importance of evaluating both mating system and contemporary gene flow for a better understanding of the biology of Anadenanthera species. This information should be considered to ensure the effective conservation and management practices of these plant species. [ABSTRACT FROM AUTHOR]