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Regenerating forests occupy large areas in the tropics, mostly as a result of deforestation for livestock and agriculture, followed by land abandonment. Despite the importance of regenerating secondary forests for tropical biodiversity conservation, studies of temporal effects of matrix regeneration on species responses in fragmented landscapes are scarce. Here, we used an Amazonian whole-ecosystem fragmentation experiment to investigate how changes in matrix quality over time through secondary forest regeneration affect bat assemblages from a functional perspective. We found that forest regeneration in the matrix positively affected functional α diversity, as well as species- and community-level functional uniqueness, reflecting an increase of species that perform different ecological functions in secondary forest over time. According to functional trait composition, animalivorous species showed the clearest …