Leaf litter plays an essential role in the functioning of forest ecosystems. They are a source of organic matter and a protective layer in forest soils as a nurturing habitat for micro- and macro-organisms. Through their successional occurrence, litter-inhabiting microfungi play a key role in litter decomposition and nutrient recycling. Despite their importance in terrestrial ecosystems, relationships of host species phylogeny and the effects of different leaf litter types on the diversity and distribution of saprobic fungi are poorly understood. The present study investigated the saprobic microfungal community structure and composition among six different host species (Dipterocarpus alatus, Nayariophyton zizyphifolium, Microcos paniculata, Afzelia xylocarpa, Dalbergia cana, and Dalbergia cultrata) located in Doi Tung, Chiang Rai, Thailand. Leaf samples were collected from each host species during three seasons, and saprobic microfungi were isolated using single spore isolation. Morphology and molecular characterizations resolved the taxonomy of the species. Multivariate analyses were performed to explore the effects of host, seasonal variations on fungal host preference and community patterns. Results indicated significant variations of saprobic microfungal distribution, diversity, and richness. Host species and seasonal variations influenced the community composition dominating in each host. The most common genera viz.Pestalotiopsis, Colletotrichum, Diaporthe, and Penicillium are dominant across all six hosts, while Apiospora, Cheatothyriales, Didymella, Ciliochorella are uncommon and showed distinct preferences for specific hosts or seasons. The study generated evidence for multiple levels of diversity and host preference of saprobes dwelling inside the leaf litter of forest ecosystems revealing their high degree of host preference.