Abstract Aims The extent and persistence of pre-Columbian human legacies in old-growth Amazonian forests are still controversial, partly because modern societies re-occupied old settlements, challenging the distinction between pre- and post-Columbian legacies. Here, we compared the effects of pre-Columbian vs. recent landscape domestication processes on soils and vegetation in two Amazonian regions. Methods We studied forest landscapes at varying distances from pre-Columbian and current settlements inside protected areas occupied by traditional and indigenous peoples in the lower Tapajós and the upper-middle Madeira river basins. By conducting 69 free-listing interviews, participatory mappings, guided-tours, 27 forest inventories, and soil analysis, we assessed the influences of pre-Columbian and current activities in soils and plant resources surrounding the settlements. Results In both regions, we found that pre-Columbian villages were more densely distributed across the landscape than current villages. Soil nutrients (mainly Ca and P) were higher closer to pre-Columbian villages but were generally not related to current villages, suggesting past soil fertilization. Soil charcoal was frequent in all forests, suggesting frequent fire events. The density of domesticated plants used for food increased in phosphorus enriched soils. In contrast, the density of plants used for construction decreased near current villages. Conclusions We detected a significant effect of past soil fertilization on food resources over extensive areas, supporting the hypothesis that pre-Columbian landscape domestication left persistent marks on Amazonian landscapes. Our results suggest that a combination of pre-Columbian phosphorus fertilization with past and current management drives plant resource availability in old-growth forests.