Multiplex Amplicon Sequencing For Microbe Identification In Community-based Culture Collections

dc.creatorLeite Armanhi
dc.creatorJaderson Silveira; Correa de Souza
dc.creatorRafael Soares; de Araujo
dc.creatorLaura Migliorini; Okura
dc.creatorVagner Katsumi; Mieczkowski
dc.creatorPiotr; Imperial
dc.creatorJuan; Arruda
dc.creatorPaulo
dc.date2016
dc.datejul
dc.date2017-11-13T13:44:10Z
dc.date2017-11-13T13:44:10Z
dc.date.accessioned2018-03-29T05:58:49Z
dc.date.available2018-03-29T05:58:49Z
dc.identifierScientific Reports. Nature Publishing Group, v. 6, p. , 2016.
dc.identifier2045-2322
dc.identifierWOS:000379396300001
dc.identifier10.1038/srep29543
dc.identifierhttp://www-nature.ez88.periodicos.capes.gov.br/articles/srep29543
dc.identifierhttp://repositorio.unicamp.br/jspui/handle/REPOSIP/328704
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/1365729
dc.descriptionMicrobiome analysis using metagenomic sequencing has revealed a vast microbial diversity associated with plants. Identifying the molecular functions associated with microbiome-plant interaction is a significant challenge concerning the development of microbiome-derived technologies applied to agriculture. An alternative to accelerate the discovery of the microbiome benefits to plants is to construct microbial culture collections concomitant with accessing microbial community structure and abundance. However, traditional methods of isolation, cultivation, and identification of microbes are time-consuming and expensive. Here we describe a method for identification of microbes in culture collections constructed by picking colonies from primary platings that may contain single or multiple microorganisms, which we named community-based culture collections (CBC). A multiplexing 16S rRNA gene amplicon sequencing based on two-step PCR amplifications with tagged primers for plates, rows, and columns allowed the identification of the microbial composition regardless if the well contains single or multiple microorganisms. The multiplexing system enables pooling amplicons into a single tube. The sequencing performed on the PacBio platform led to recovery near-full-length 16S rRNA gene sequences allowing accurate identification of microorganism composition in each plate well. Cross-referencing with plant microbiome structure and abundance allowed the estimation of diversity and abundance representation of microorganism in the CBC.
dc.description6
dc.descriptionRepsol
dc.descriptionRepsol Sinopec
dc.languageEnglish
dc.publisherNature Publishing Group
dc.publisherLondon
dc.relationScientific Reports
dc.rightsaberto
dc.sourceWOS
dc.subjectPlant Microbiome
dc.subjectRoot Microbiota
dc.subjectRhizosphere
dc.subjectBacteria
dc.subjectProject
dc.subjectDna
dc.titleMultiplex Amplicon Sequencing For Microbe Identification In Community-based Culture Collections
dc.typeArtículos de revistas


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