Detection of leaf-associated fungi based on PCR and nucleotide sequence ofthe ribosomal internal transcribed spacer (ITS) in Miscanthus

PCR amplification of the internal transcribed spacer region (ITS) of the nu clear ribosomal DNA from the genomic DNA isolated from the leaf tissue of M iscanthus sinensis varieties and M. floridulus resulted in two amplified fr agments. Via FASTA search, the shorter length of about 500 base pairs was i dentified as the ITS region of fungi symbiotic in the grass foliage. Two Cl adosporium species, one Fusarium and one basidiomycete were added to the my coflora of Miscanthus grassland. Using a Southern blotting and hybridizatio n, fungal ITS was also detected from the genomic DNAs isolated from sterili zed seedling, a suggestive seed transmission of endophytic fungi. The "cont aminated" genomic DNA may be a hindrance to the phylogenetic analysis of Mi scanthus itself, but may provide other insights into the biodiversity of gr assland ecosystems. The endophytic foliage fungi were not detected in M. si nensis var. condensatus, probably a result of its apomictic reproduction.