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

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 

作者机构：Department of Life SciencesNational Cheng Kung UniversityTainanTaiwan 701 Institute of Tropical Plant SciencesNational Cheng Kung UniversityTainanTaiwan 701 Department of Engineering Science and Supercomputing Research CenterNational Cheng Kung UniversityTainanTaiwan 701 

出 版 物：《Fungal Diversity》 (真菌多样性（英文）)

年 卷 期：2014年第66卷第3期

页      面：77-88页

核心收录：

学科分类：090706[农学-园林植物与观赏园艺] 0907[农学-林学] 09[农学] 

基　　金：This study was financially supported by the National Cheng Kung University and the National Science Council Taiwan 

主　　题：DNA barcode Metagenomics Mycorrhizal fungi Next-generation sequencing Phalaenopsis orchid Rank scoring 

摘      要：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.