Step‐by‐Step Pipeline for the Ecological Analysis of Endophytic Fungi using ITS nrDNA Data

Step‐by‐Step Pipeline for the Ecological Analysis of Endophytic Fungi using ITS nrDNA Data
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使用 ITS nrDNA 数据对内生真菌进行生态分析的逐步流程

DOI:
10.1002/cpmc.96
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发表时间:
2020
影响因子:
--
通讯作者:
Chaverri, Priscila
Chaverri, Priscila
中科院分区:
--
文献类型:
--
作者:
Montero‐Vargas, Maripaz;Escudero‐Leyva, Efraín;Díaz‐Valerio, Stefani;Chaverri, Priscila

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The nuclear ribosomal DNA internal transcribed spacer (ITS) is accepted as the genetic marker or barcode of choice for the identification of fungal samples. Here, we present a protocol to analyze fungal ITS data, from quality preprocessing of raw sequences to identification of operational taxonomic units (OTUs), taxonomic classification, and assignment of functional traits. The pipeline relies on well‐established and manually curated data collections, namely the UNITE database and the FUNGuild script. As an example, real ITS data from culturable endophytic fungi were analyzed, providing detailed descriptions for every step, parameter, and downstream analysis, and finishing with a phylogenetic analysis of the sequences and assigned ecological roles. This article constitutes a comprehensive guide for researchers that have little familiarity with bioinformatic analysis of essential steps required in further ecological studies of fungal communities. © 2020 by John Wiley & Sons, Inc.Basic Protocol 1: Raw sequencing data processingSupport Protocol: Building a BLAST databaseBasic Protocol 2: Obtaining information from databasesBasic Protocol 3: Phylogenetic analysis
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