The aim of this study was to explore the diversity of macrofungi in the nature trail of Namtok Phlio National Park, Chanthaburi Province, Thailand, the role in the ecology, and their edibility. Fifty-eight macrofungi samples were collected from the nature trail in June 2023. Twenty-eight macrofungi samples with different fruiting bodies (a similar morphology was excluded) were selected and classified according to their morphology. The results indicated that the gilled fungi were the most diverse species (42.9%). Subsequently, they were identified by sequence analysis of the internal transcribed spacer (ITS). They were classified into 2 phyla, 3 classes, 9 orders, 18 families, and 23 genera, and most commonly in the phylum Basidiomycota (89.3%), the family Polyporaceae (21.4%), and played a major role as saprotrophs (85.7%). Additionally, most of them did not have any information for the edibility data (82.1%), however, edible (10.7%) and poisonous (7.1%) macrofungi were reported. Moreover, some macrofungi samples need further investigation for molecular identification based on analysis of additional genes. These results could be used as a database of the macrofungi diversity in the nature trail of Namtok Phlio National Park.
