Previous research has shown that ancient Maya 'forest gardens'-tree-dominated home gardens containing a diversity of tree species used for daily household needsstill resonate in the species composition of the modern forest. Centuries of positive interspecies interactions may enhance the reproductive and survival success of garden species selected and encouraged by experienced Maya forest gardeners. We hypothesized that such interaction may result in aggregated spatial patterns between 32 pre-selected, commonly utilized forest garden species. In this paper, we developed a novel randomization (Monte Carlo) method designed to measure and test if the spatial relationships among pairs of Maya 'forest garden' trees species differ between areas that experienced high and low ancient settlement density in northwestern Belize. A total of 28 high ancient settlement density and 27 low ancient settlement density plots containing a total of 2772 and 3134 trees, respectively, were used for this study. The analysis revealed that 58 pairs of forest garden species tended to 'cluster' together significantly more often in the high settlement density areas than would be expected in a random distribution. In low settlement density plots, only 12 pairs of species exhibited a significant clustered spatial relationship. The effect was not species specific, suggesting that some synergistic relationships, mediated by third-party agents such as dispersers, may occur at the community level. The impacts of ancient human land use, echoing across centuries of dispersal, colonization, disturbance, and biotic and abiotic interactions, can have important implications for understanding the current biodiversity patterns and processes.Abstract in Spanish is available at http://www.blackwell-synergy.com/loi/btp.