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 | Acesso ao texto completo restrito à biblioteca da Embrapa Soja. Para informações adicionais entre em contato com valeria.cardoso@embrapa.br. |
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Registro Completo |
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Biblioteca(s): |
Embrapa Soja. |
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Data corrente: |
17/12/2015 |
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Data da última atualização: |
05/08/2017 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Autoria: |
KUMA, K. M.; LOPES-CAITAR, V. S.; ROMERO, C. C. T.; SILVA, S. M. H.; CARVALHO, M. C. C. G.; ABDELNOOR, R. V.; DIAS, W. P.; MARCELINO-GUIMARÃES, F. C. |
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Afiliação: |
UEL; UEL; CNPSo; UEL; UENP; RICARDO VILELA ABDELNOOR, CNPSO; WALDIR PEREIRA DIAS, CNPSO; FRANCISMAR CORREA MARCELINO GUIMARÃES, CNPSO. |
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Título: |
A high efficient protocol for soybean root transformation by Agrobacterium rhizogenes and most stable reference genes for RT-qPCR analysis. |
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Ano de publicação: |
2015 |
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Fonte/Imprenta: |
Plant Cell Reports, v. 34, n. 11, p. 1987-2000, 2015. |
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ISSN: |
1432-203X |
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DOI: |
10.1007/s00299-015-1845-2 |
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Idioma: |
Inglês |
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Conteúdo: |
Gene functional analyses are essential to the validation of results obtained from in silico and/or geneprospecting studies. Genetic transformation methods that yield tissues of transient expression quickly have been of considerable interest to researchers. Agrobacterium rhizogenes-mediated transformation methods, which are employed to generate plants with transformed roots, have proven useful for the study of stress caused by root phytopathogens via gene overexpression and/or silencing. While some protocols have been adapted to soybean plants, transformation efficiencies remain limited; thus, few viable plants are available for performing bioassays. Furthermore, mRNA analyses that employ reverse transcription quantitative polymerase chain reactions (RT-qPCR) require the use of reference genes with stable expression levels across different organs, development steps and treatments. In the present study, an A. rhizogenes-mediated soybean root transformation approach was optimized. The method delivers significantly higher transformation efficiency levels and rates of transformed plant recovery, thus enhancing studies of soybean abiotic conditions or interactions between phytopathogens, such as nematodes. A 55 % transformation efficiency was obtained following the addition of an acclimation step that involves hydroponics and different selection processes. The present study also validated the reference genes GmELF1-b and GmELF1-a as the most stable to be used in RT-qPCR analysis in composite plants, mainly under nematode infection. MenosGene functional analyses are essential to the validation of results obtained from in silico and/or geneprospecting studies. Genetic transformation methods that yield tissues of transient expression quickly have been of considerable interest to researchers. Agrobacterium rhizogenes-mediated transformation methods, which are employed to generate plants with transformed roots, have proven useful for the study of stress caused by root phytopathogens via gene overexpression and/or silencing. While some protocols have been adapted to soybean plants, transformation efficiencies remain limited; thus, few viable plants are available for performing bioassays. Furthermore, mRNA analyses that employ reverse transcription quantitative polymerase chain reactions (RT-qPCR) require the use of reference genes with stable expression levels across different organs, development steps and treatments. In the present study, an A. rhizogenes-mediated soybean root transformation approach was optimized. The method delivers significantly higher transformation efficiency levels and rates of transformed plant recovery, thus enhancing studies of soybean abiotic conditions or interactions between phytopathogens, such as nematodes. A 55 % transformation efficiency was obtained following the addition of an acclimation step that involves hydroponics and different selection processes. The present study also validated the reference genes GmELF1-b and GmELF1-a as the most stable to be used in RT-qPCR analysis i... Mostrar Tudo |
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Thesagro: |
Soja. |
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Categoria do assunto: |
-- |
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Marc: |
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024 7 $a10.1007/s00299-015-1845-2$2DOI
100 1 $aKUMA, K. M.
245 $aA high efficient protocol for soybean root transformation by Agrobacterium rhizogenes and most stable reference genes for RT-qPCR analysis.$h[electronic resource]
260 $c2015
520 $aGene functional analyses are essential to the validation of results obtained from in silico and/or geneprospecting studies. Genetic transformation methods that yield tissues of transient expression quickly have been of considerable interest to researchers. Agrobacterium rhizogenes-mediated transformation methods, which are employed to generate plants with transformed roots, have proven useful for the study of stress caused by root phytopathogens via gene overexpression and/or silencing. While some protocols have been adapted to soybean plants, transformation efficiencies remain limited; thus, few viable plants are available for performing bioassays. Furthermore, mRNA analyses that employ reverse transcription quantitative polymerase chain reactions (RT-qPCR) require the use of reference genes with stable expression levels across different organs, development steps and treatments. In the present study, an A. rhizogenes-mediated soybean root transformation approach was optimized. The method delivers significantly higher transformation efficiency levels and rates of transformed plant recovery, thus enhancing studies of soybean abiotic conditions or interactions between phytopathogens, such as nematodes. A 55 % transformation efficiency was obtained following the addition of an acclimation step that involves hydroponics and different selection processes. The present study also validated the reference genes GmELF1-b and GmELF1-a as the most stable to be used in RT-qPCR analysis in composite plants, mainly under nematode infection.
650 $aSoja
700 1 $aLOPES-CAITAR, V. S.
700 1 $aROMERO, C. C. T.
700 1 $aSILVA, S. M. H.
700 1 $aCARVALHO, M. C. C. G.
700 1 $aABDELNOOR, R. V.
700 1 $aDIAS, W. P.
700 1 $aMARCELINO-GUIMARÃES, F. C.
773 $tPlant Cell Reports$gv. 34, n. 11, p. 1987-2000, 2015.
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Registro original: |
Embrapa Soja (CNPSO) |
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| Registros recuperados : 12 | |
| 3. |  | RANDIG, O.; CARNEIRO, R. M. D. G.; CASTAGNONE SERENO, P. Identificação das principais espécies de Melodogyne parasitas do cafeeiro no Brasil com marcadores scar-café em multiplex-PCR. Fitopatologia Brasileira, Brasília, DF, v. 29, supl., p. S81, ago. 2004. Edição dos Resumos do XXXVII Congresso Brasileiro de Fitopatologia, Gramado, RS, ago. 2004.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Recursos Genéticos e Biotecnologia. |
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| 4. | | CARNEIRO, R. M. D. G.; RANDIG, O.; ALMEIDA, M. R. A.; GONÇALVES, W. Identificação e caracterização em Melodogyne spp. em cafeeiro nos estados de São Paulo e Minas Gerais através dos fenótipos das esterases e scar.multiplex-PCR. Fitopatologia Brasileira, Brasília, DF, v. 29, supl., p. S82, ago. 2004. Edição dos Resumos do XXXVII Congresso Brasileiro de Fitopatologia, Gramado, RS, ago. 2004.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Recursos Genéticos e Biotecnologia. |
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| 5. | | MOTA, F. C.; CARNEIRO, R. M. D. G.; MARTINS, I.; RANDIG, O. Estabelecimento de uma coleção criopreservada de Meloidogyne SPP. In: ENCONTRO DO TALENTO ESTUDANTIL DA EMBRAPA RECURSOS GENÉTICOS E BIOTECNOLOGIA, 10., 2005, Brasília, DF. Anais: resumos dos trabalhos. Brasília, DF: Embrapa Recursos Genéticos e Biotecnologia, 2005. p. 199.| Biblioteca(s): Embrapa Recursos Genéticos e Biotecnologia. |
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| 6. | | SANTOS, M. F. A.; RANDIG, O.; CARNEIRO, R. M. D. G. Uso de marcadores scar para identificação de Meloidogyne incognita, M. javanica e M. arenaria. In: ENCONTRO DO TALENTO ESTUDANTIL DA EMBRAPA RECURSOS GENÉTICOS E BIOTECNOLOGIA, 10., 2005, Brasília, DF. Anais: resumos dos trabalhos. Brasília, DF: Embrapa Recursos Genéticos e Biotecnologia, 2005. p. 216.| Biblioteca(s): Embrapa Recursos Genéticos e Biotecnologia. |
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| 7. | | SOUZA, H. J. M.; RANDIG, O.; CARNEIRO, R. M. D. G. Uso de marcadores scar-rapd em multiplex-pcr para detecção de espécies de Meloidogyne, parasitas do cafeeiro. ENCONTRO DO TALENTO ESTUDANTIL DA EMBRAPA RECURSOS GENÉTICOS E BIOTECNOLOGIA, 8., 2003, Brasília, DF. Anais: resumos dos trabalhos. Brasília, DF: Embrapa Recursos Genéticos e Biotecnologia, 2003. p. 53| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Recursos Genéticos e Biotecnologia. |
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| 9. | | RANDIG, O.; DEAU, F.; SANTOS, M. F. A. dos; TIGANO, M. S.; CARNEIRO, R. M. D. G.; CASTAGNONE-SERENO, P. A novel species-specific satellite DNA family in the invasive root-knot nematode Meloidogyne mayaguensis and its potential use for diagnostics. In: INTERNATIONAL CONGRESS OF TROPICAL NEMATOLOGY, 2., 2009, Maceió. Abstracts... Maceió: ONTA: SBN, 2009. 1 CD-ROM.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Recursos Genéticos e Biotecnologia. |
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| 10. | | SANTOS, M. F. A.; TIGANO, M. S.; ALMEIDA, M. R. A.; RANDIG, O.; CARNEIRO, R. M. D. G. Variabilidade genética de populações de Meloidogyne arenaria através da análise de marcadores "RAPD". Nematologia Brasileira, v. 30, n. 1, p. 129, 2006. Resumos apresentados no CONGRESSO BRASILEIRO DE NEMATOLOGIA, 26., 2006, Campos de Goytacazes, Rio de Janeiro.| Tipo: Resumo em Anais de Congresso | Circulação/Nível: -- - -- |
| Biblioteca(s): Embrapa Recursos Genéticos e Biotecnologia. |
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| 11. | | ALMEIDA, C. D. S. de; MAGALHÃES, J. C. C.; FRANCO, P.; SALOMÃO, D.; RANDIG, O.; CARNEIRO, R. M. D. G.; GROSSI-de-SÁ, M. F.; ROCHA, T. L. Efeito de extratos de sementes de plantas antagonistas sobre a mortalidade de juvenis de segundo estágio de Meloidogine incognita. In: ENCONTRO DO TALENTO ESTUDANTIL DA EMBRAPA RECURSOS GENÉTICOS E BIOTECNOLOGIA, 9., 2004, Brasília, DF. Anais: resumos dos trabalhos. Brasília, DF: Embrapa Recursos Genéticos e Biotecnologia, 2004. p. 128.| Biblioteca(s): Embrapa Recursos Genéticos e Biotecnologia. |
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| 12. | | TIGANO, M. S.; CASTAGNONE SERENO, P.; QUEIROZ, P. R.; SIQUEIRA, K. M. S.; TEIXEIRA, C. C.; SANTOS, M. F. A.; ALMEIDA, M. R. A.; RANDIG, O.; CARNEIRO, R. M. D. G. Molecular markers for specific diagnostic and genetic variability of Meloidogyne mayaguensis isolates. In: INTERNATIONAL CONGRESS OF TROPICAL NEMATOLOGY, 2., 2009, Maceió. Abstracts... Maceió: ONTA: SBN, 2009. 1 CD-ROM.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Recursos Genéticos e Biotecnologia. |
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| Registros recuperados : 12 | |
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