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The maintenance of seedling diversity of animal-dispersed tree species is fundamental for the structure and function of forest patches in fragmented tropical rainforests. Nonetheless, the effects of landscape structure at different spatial scales on α- and β-diversity of tree seedling communities are recently explored. Using a multi-scale approach, we assessed the relative effect of landscape composition and configuration on α- and β-diversity of animal-dispersed seedlings within 16 forest patches in the Lacandona rainforest, Mexico. We assessed these effects at 13 spatial scales (from 300 to 1500 m radius, at 100 m intervals) for three metrics of effective number of species considering α- and β-diversity. We found that α-diversity was largely affected by landscape composition and β-diversity by landscape configuration. On the one hand, the amount of secondary forest influenced α-diversity. Additionally, species richness increased in landscapes with highly aggregated forest patches. On the other hand, β-diversity was affected positively by forest fragmentation and negatively by the edge contrast of forest patches with the surrounding matrix. Our findings indicate that landscape configuration is a strong driver of seedling diversity in highly deforested rainfor ests. Promoting forest patches and secondary forests through payment for ecosystem services’ programs, favoring matrix quality within land-sharing schemes of smallholder agriculture and secondary forest management, and identifying restoration opportunities for assisted or unassisted natural regeneration are urgently needed for conservation of seed ling diversity in human-modified tropical landscapes. Inglés