A ribosomal RNA gene intergenic spacer based PCR and DGGE fingerprinting method for the analysis of specific rhizobial communities in soil

dc.creatorde Oliveira, VM
dc.creatorManfio, GP
dc.creatorCoutinho, HLD
dc.creatorKeijzer-Wolters, AC
dc.creatorvan Elsas, JD
dc.date2006
dc.dateMAR
dc.date2014-11-17T06:25:20Z
dc.date2015-11-26T16:57:58Z
dc.date2014-11-17T06:25:20Z
dc.date2015-11-26T16:57:58Z
dc.date.accessioned2018-03-28T23:45:36Z
dc.date.available2018-03-28T23:45:36Z
dc.identifierJournal Of Microbiological Methods. Elsevier Science Bv, v. 64, n. 3, n. 366, n. 379, 2006.
dc.identifier0167-7012
dc.identifierWOS:000235745000007
dc.identifier10.1016/j.mimet.2005.05.015
dc.identifierhttp://www.repositorio.unicamp.br/jspui/handle/REPOSIP/53702
dc.identifierhttp://www.repositorio.unicamp.br/handle/REPOSIP/53702
dc.identifierhttp://repositorio.unicamp.br/jspui/handle/REPOSIP/53702
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/1277817
dc.descriptionA direct molecular method for assessing the diversity of specific populations of rhizobia in soil, based on nested PCR amplification of 16S-23S ribosomal RNA gene (rDNA) intergenic spacer (IGS) sequences, was developed. Initial generic amplification of bacterial rDNA IGS sequences from soil DNA was followed by specific amplification of (1) sequences affiliated with Rhizobium leguminosarum "sensu lato" and (2) R. tropici. Using analysis of the amplified sequences in clone libraries obtained on the basis of soil DNA, this two-sided method was shown to be very specific for rhizobial subpopulations in soil. It was then further validated as a direct fingerprinting tool of the target rhizobia based on denaturing gradient gel electrophoresis (DGGE). The PCR-DGGE approach was applied to soils from fields in Brazil cultivated with common bean (Phaseolus vulgaris) under conventional or no-tillage practices. The community fingerprints obtained allowed the direct analysis of the respective rhizobial community structures in soil samples from the two contrasting agricultural practices. Data obtained with both primer sets revealed clustering of the community structures of the target rhizobial types along treatment. Moreover, the DGGE profiles obtained with the R. tropici primer set indicated that the abundance and diversity of these organisms were favoured under NT practices. These results suggest that the R. leguminosarum-as well as R. tropici-targeted IGS-based nested PCR and DGGE are useful tools for monitoring the effect of agricultural practices on these and related rhizobial subpopulations in soils. (C) 2005 Elsevier B.V All rights reserved.
dc.description64
dc.description3
dc.description366
dc.description379
dc.languageen
dc.publisherElsevier Science Bv
dc.publisherAmsterdam
dc.publisherHolanda
dc.relationJournal Of Microbiological Methods
dc.relationJ. Microbiol. Methods
dc.rightsfechado
dc.rightshttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dc.sourceWeb of Science
dc.subjectculture-independent analysis
dc.subjectrhizobia
dc.subjectrDNA spacer
dc.subjectsoil management
dc.subjectdiversity
dc.subjectDGGE
dc.subjectGradient Gel-electrophoresis
dc.subjectPolymerase Chain-reaction
dc.subjectPaenibacillus-azotofixans
dc.subjectMicrobial Diversity
dc.subjectDna Extraction
dc.subjectHost Plants
dc.subjectLeguminosarum
dc.subjectStrains
dc.subjectAmplification
dc.subjectTropici
dc.titleA ribosomal RNA gene intergenic spacer based PCR and DGGE fingerprinting method for the analysis of specific rhizobial communities in soil
dc.typeArtículos de revistas


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