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Deciphering mycorrhizal fungi in cultivated Phalaenopsis microbiome with next-generation sequencing of multiple barcodes

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Fungal Diversity 2014年第3期66卷 77-88页

作者： Chao-Li Huang Feng-Yin Jian Hao-Jen Huang Wen-Chi Chang Wen-Luan Wu Chi-Chuan Hwang Ruey-Hua Lee Tzen-Yuh Chiang Department of Life Sciences National Cheng Kung UniversityTainanTaiwan 701 Institute of Tropical Plant Sciences National Cheng Kung UniversityTainanTaiwan 701 Department of Engineering Science and Supercomputing Research Center National Cheng Kung UniversityTainanTaiwan 701

Identifying the species composition of a microbial ecosystem is often hampered by difficulties in culturing the organisms and in the low sequencing depth of traditional DNA *** analysis,a huge-scale nucleotide-sequence-based tool,can overcome such *** this study,Sanger sequencing of 500 nrITS clones uncovered 29 taxa of 19 fungal genera,whereas metagenomics with next-generation sequencing identified 512 operational taxonomic units(OTUs)for ITS1/2 and 364 for ITS3/***,high throughput sequencing of PCR amplicons of ITS1/2,ITS3/4,nrLSU-LR,nrLSU-U,mtLSU,and mtATP6,all with at least 1,300×coverage and about 21 million reads in total,yielded a very diverse fungal *** fact that 74%of the OTUs were exclusively uncovered with single barcodes indicated that each marker provided its own insights into the fungal *** deal with the high heterogeneity in the data and to integrate the information on species composition across barcodes,a rank-scoring strategy was ***,205 genera among 64 orders of fungi were identified in healthy Phalaenopsis *** the barcodes utilized,ITS1/2,ITS3/4,and nrLSU-U were the most competent in uncovering the fungal *** barcodes,though detecting different compositions likely due to primer preference,provided complementary and comprehensive power in deciphering the microbial diversity,especially in revealing rare species.

关键词： DNA barcode Metagenomics Mycorrhizal fungi Next-generation sequencing Phalaenopsis orchid rank scoring

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