The endophytic fungal diversity in tropical forests has not been completely investigated despite the anticipated high diversity. Especially, information on the diversity of endophytic fungi associated with tree leaves is lacking. This study is the first to conduct a metagenomic analysis with Next-generation sequencing to examine the putative hyper-diversity of endophytic fungi in two tropical forests in Malaysia and Australia and one subtropical forest in Japan. At each site, a total of six live leaves, three leaves per each of two dominant tree species, were sampled. Fungal DNA was extracted and ITS-1 region of rDNA was sequenced with a Roch 454 Titanium.

A total of 833 operational taxonomic units (95% similarity) were recorded, with 95-230 OTUs per tree. Possibly, the result suggested the hyper-diversity of endophytic fungi associated with tree leaves in tropical forests.