Abstract
BACKGROUND: The variation in fungal community composition within a single habitat space has been extensively studied in forest ecosystems. However, the spatial and temporal distribution of fungi across contiguous habitats, particularly at a local scale and in tropical regions, remains underexplored. In this study, we examined the fungal community composition across multiple habitats proximal to each other over two seasons in seven Fagaceae species in Taiwanese broadleaf forests. We tested how local spatial scale and habitat influence community assembly. RESULTS: Using a metabarcoding approach, we sequenced ITS2 regions from 864 samples collected from four distinct habitats-leaves, twigs, litter, and soil. We identified 11,600 fungal amplicon sequence variants (ASVs), with community composition differing significantly between habitats proximal to each other. Generalized dissimilarity modeling (GDM) revealed that spatial distance, interacting with precipitation, was the strongest predictor of fungal turnover, particularly in the phyllosphere. Normalized Stochasticity Ratio (NST) analyses further highlighted contrasting assembly processes, with deterministic influences dominating in the phyllosphere habitat, while stochasticity prevailed in soil and litter. Random forest analysis accurately classified habitats based on ASVs' relative abundances, with strong predictors were mostly habitat-specific ASVs prevalent in soil. Misclassified samples were due to secondary contact of fungi between adjacent habitats. Co-occurrence network analysis revealed more complex and deterministic networks in leaf and twig habitats, while soil was driven by stochastic processes and contained most habitat-specific ASVs. A Cladosporium sp. emerged as a keystone species, maintaining network stability across forests. CONCLUSION: This study reveals how local spatial variation and habitat shape distinct fungal communities in tropical forests, with deterministic processes dominating in some habitats and stochasticity playing a key role in others. We show extremely high turnover in fungal community are present over very short distances and that local fungal taxa are strong habitat predictors. These findings highlight the importance of studying coexisting habitats to gain a deeper understanding of fungal biogeography and ecosystem function.