Detection of Leaf-associated Fungi Based on PCR and Nucleotide Sequence of the Ribosomal Internal Transcribed Spacer (ITS) in Miscanthus

利用PCR擴充及分子序列核核糖體DNA ITS片段偵測芒屬植物葉共生的菌類

江友中(Yu-Chung Chiang) ； 周昌弘(Chang-Hung Chou) ； 李佩叡(Pei-Ray Lee) ； 蔣鎮宇(Tzen-Yuh Chiang)

《Botanical Bulletin of Academia Sinica》 42卷1期 (2001/01) Pp. 39-44

https://doi.org/10.7016/BBAS.200101.0039

摘要

由中國芒各變種及五節芒葉組織萃取的基因體DNA，利用PCR擴充核核糖體DNA ITS發現兩個大小不同的片段，以FASTA搜尋，其中較小約500bp的片段，鑑定其較可能來自此禾草葉片中共生的菌類，其中包含二種Cladosporium，一種fusarium及一個basidiomycete。利用南方墨點雜合反應，此菌類ITS片段亦存在於經消毒處理的小苗，指示此菌類可經由種子傳遞。此一“污染的”基因體DNA對於芒屬植物的親緣重建可能造成障礙，另一方面卻提供我們了解草原生態系多樣性的訊息。內生菌并未在八丈芒中被偵測到，可能與其無配生殖有極大相關。

關鍵字

污染 ； 葉組織共生菌 ； 中國芒 ； 菌類誌 ； 親緣 ； 核糖體DNA ITS

並列摘要

PCR amplification of the internal transcribed spacer region (ITS) of the nuclear ribosomal DNA from the genomic DNA isolated from the leaf tissue of Miscanthus sinensis varieties and M. floridulus resulted in two amplified fragments. Via FASTA search, the shorter length of about 500 base pairs was identified as the ITS region of fungi symbiotic in the grass foliage. Two Cladosporiuin species, one Fusarium and one basidiomycete were added to the mycoflora of Miscanthus grassland. Using a Southern blotting and hybridization, fungal ITS was also detected from the genomic DNAs isolated from sterilized seedling, a suggestive seed transmission of endophytic fungi. The “contaminated” genomic DNA may be a hindrance to the phylogenetic analysis of Miscanthus itself, but may provide other insights into the biodiversity of grassland ecosystems. The endophytic foliage fungi were not detected in M. sinensis var. condensatus, probably a result of its apomictic reproduction.

並列關鍵字

Contamination ； Foliage fungi ； Miscanthus sinensis ； Mycoflora ； Phylogeny ； rDNA ITS