Kanchanaburi forests are well known for high diversity of Termitomyces, an un-culturable and
economic mushroom in Thailand, but their systematics are limited and unorganized. We, therefore,
identified 28 Termitomyces isolates collected from 8 districts in Kanchanaburi province based on
morphological characteristics and ITS1-5.8S-ITS2 rDNA sequences. Nine species were identified
as T. albiceps, T. bulborhizus, T. cylindricus, T. heimii, T. microcarpus, T. radicatus, T.
entolomoides, T. fuliginosus, and T. clypeatus. Analysis of ITS1-5.8S-ITS2 rDNA sequences of
these Termitomyces species revealed that morphological characteristics of T. clypeatus represented
the most extremely variations that had not been described in any identification references. The
inferred Neighbor-Joining phylogram of ITS1-5.8S-ITS2 rDNA sequences showed that 13
selected Termitomyces isolates were monophyletic and diverged into 2 clades with no common
characteristic that can be shared in each clade. In addition, the phylogenetic study demonstrated
the monophyletic tree from pure Kanchanaburi Termitomyces isolates and mixture of Asia and
African Termitomyces samples implied that both Asia and African Termitomyces species have
evolved from the same ancestor.