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Biblioteca(s): |
Embrapa Solos. |
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Data corrente: |
20/09/2021 |
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Data da última atualização: |
20/09/2021 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Autoria: |
OLIVEIRA, V. M. de; MANFIO, G. P.; COUTINHO, H. L. da C.; KEIJZER-WOLTERS, A. C.; ELSAS, J. D. van. |
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Afiliação: |
VALÉRIA MAIA DE OLIVEIRA, UNICAMP; GILSON PAULO MANFIO, UNICAMP; HEITOR LUIZ DA COSTA COUTINHO, CNPS; ANNEKE CHRISTINA KEIJZER-WOLTERS, Plant Research International; JAN DIRK VAN ELSAS, University of Groningen. |
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Título: |
A ribosomal RNA gene intergenic spacer based PCR and DGGE fingerprinting method for the analysis of specific rhizobial communities in soil. |
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Ano de publicação: |
2006 |
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Fonte/Imprenta: |
Journal of Microbiological Methods, v. 64, n. 3, p. 366-379, Mar. 2006. |
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DOI: |
https://doi.org/10.1016/j.mimet.2005.05.015 |
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Idioma: |
Inglês |
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Conteúdo: |
A 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. MenosA 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 us... Mostrar Tudo |
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Palavras-Chave: |
Culture-independent analysis; DGGE; Diversity; RDNA spacer; Rhizobia. |
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Thesaurus Nal: |
Soil management. |
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Categoria do assunto: |
P Recursos Naturais, Ciências Ambientais e da Terra |
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Marc: |
LEADER 02461naa a2200253 a 4500
001 2134556
005 2021-09-20
008 2006 bl uuuu u00u1 u #d
024 7 $ahttps://doi.org/10.1016/j.mimet.2005.05.015$2DOI
100 1 $aOLIVEIRA, V. M. de
245 $aA ribosomal RNA gene intergenic spacer based PCR and DGGE fingerprinting method for the analysis of specific rhizobial communities in soil.$h[electronic resource]
260 $c2006
520 $aA 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.
650 $aSoil management
653 $aCulture-independent analysis
653 $aDGGE
653 $aDiversity
653 $aRDNA spacer
653 $aRhizobia
700 1 $aMANFIO, G. P.
700 1 $aCOUTINHO, H. L. da C.
700 1 $aKEIJZER-WOLTERS, A. C.
700 1 $aELSAS, J. D. van
773 $tJournal of Microbiological Methods$gv. 64, n. 3, p. 366-379, Mar. 2006.
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| 20. |  | OLIVEIRA, V. M. de; COUTINHO, H. L. da C.; ELSAS, J. D. van; MANFIO, G. P. Avaliacao da diversidade de rizobios em amostras ambientais atraves do uso de primers especificos. In REUNIAO BRASILEIRA DE FERTILIDADE DE SOLO E NUTRICAO DE PLANTAS, 23.; REUNIAO BRASILEIRA SOBRE MICORRIZAS, 7.; SIMPOSIO BRASILEIRO DE MICROBIOLOGIA DO SOLO, 5.; REUNIAO BRASILEIRA DE BIOLOGIA DO SOLO, 2., 1998, Caxambu,MG. Resumos... Lavras: UFLA / SBCS / SBM, 1998. p.808. FERTBIO 98: Interrelacao fertilidade, biologia de solo e nutricao de plantas: consolidacao de um paradigma.| Biblioteca(s): Embrapa Solos. |
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