The leaf surface, known as the phylloplane, presents an oligotrophic and heterogeneous environment due to its topography and uneven distribution of resources. Although it is a challenging environment, leaves support abundant bacterial communities that are spatially structured. However, the factors influencing these spatial distribution patterns are not well understood. To study the changes in population density and spatial distribution of bacteria in synthetic communities, the behaviour of two common bacterial groups in the Arabidopsis thaliana leaf microbiota—Methylobacterium (methylobacteria) and Sphingomonas (sphingomonads)—was examined. Using synthetic communities consisting of two or three species, the hypothesis was tested that the presence of a third species affects the density and spatial interaction of the other two species. Results indicated that methylobacteria exhibit greater sensitivity to changes in population densities and spatial patterns, with higher intra-genus competition and lower densities and aggregation compared to sphingomonads. Pairwise comparisons were insufficient to explain the shifts observed in three-species communities, suggesting that higher-order interactions influence the structuring of complex communities. This emphasises the role of multispecies interactions in determining spatial patterns and community dynamics on the phylloplane.
