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 | Acesso ao texto completo restrito à biblioteca da Embrapa Agrobiologia. Para informações adicionais entre em contato com cnpab.biblioteca@embrapa.br. |
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Registro Completo |
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Biblioteca(s): |
Embrapa Agrobiologia. |
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Data corrente: |
09/11/2021 |
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Data da última atualização: |
09/11/2021 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Autoria: |
SOARES, I. C.; PACHECO, R. S.; SILVA, C. G. N. da; SANTOS, R. S.; BALDANI, J. I.; URQUIAGA, S.; VIDAL, M. S.; ARAUJO, J. L. S. de. |
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Afiliação: |
ISIS CAPELLA SOARES, UFRRJ; RAFAEL SANCHES PACHECO, UFRRJ; CLEUDISON GABRIEL NASCIMENTO DA SILVA, UFLA; RAFAEL SALAZAR SANTOS, UFRRJ; JOSE IVO BALDANI, CNPAB; SEGUNDO SACRAMENTO U CABALLERO, CNPAB; MARCIA SOARES VIDAL, CNPAB; JEAN LUIZ SIMOES DE ARAUJO, CNPAB. |
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Título: |
Real-time PCR method to quantify Sp245 strain of Azospirillum baldaniorum on Brachiaria grasses under field conditions. |
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Ano de publicação: |
2021 |
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Fonte/Imprenta: |
Plant and Soil, 06 September 2021 |
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ISSN: |
0032-079X |
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DOI: |
https://doi.org/10.1007/s11104-021-05137-y |
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Idioma: |
Inglês |
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Conteúdo: |
Bacterial quantification by qPCR is considered the gold standard for microbial molecular diagnosis. However, a fundamental pre-requisite in this methodology is the designing of specific primers for the bacterium of interest. With the increase in bacterial genome sequencing data in the recent years, it has become possible to design specific primers that can be used to quantify different strains of the same bacterial species. Methods: To develop a real-time PCR (qPCR) protocol for the specific quantification of Azospirillum baldaniorum Sp245 strain (old Azospirillum brasilense), the Sp245 genome sequence was fragmented into small contigs with 500 base pairs each, and analyzed for similarity against the NCBI non-redundant database. A. baldaniorum-specific contigs were used to design the primers. The best pair of primers was used to quantify these bacteria after inoculation in different cultivars of Brachiaria, grown under field conditions. Results: Our results showed that the primer pair Sp245p10 was highly specific for the Sp245 strain in the Brachiaria root and shoot field under different conditions. The qPCR assay using these primers showed differences among cultivars in the number of bacteria detected in plants after inoculation. Additionally, the number of bacteria observed in the roots was higher than that in the shoots. Conclusion: The qPCR methodology using a Sp245 strain-specific primer may be used to monitor A. baldaniorum inoculated into other plants and may find potential application in field experiments. MenosBacterial quantification by qPCR is considered the gold standard for microbial molecular diagnosis. However, a fundamental pre-requisite in this methodology is the designing of specific primers for the bacterium of interest. With the increase in bacterial genome sequencing data in the recent years, it has become possible to design specific primers that can be used to quantify different strains of the same bacterial species. Methods: To develop a real-time PCR (qPCR) protocol for the specific quantification of Azospirillum baldaniorum Sp245 strain (old Azospirillum brasilense), the Sp245 genome sequence was fragmented into small contigs with 500 base pairs each, and analyzed for similarity against the NCBI non-redundant database. A. baldaniorum-specific contigs were used to design the primers. The best pair of primers was used to quantify these bacteria after inoculation in different cultivars of Brachiaria, grown under field conditions. Results: Our results showed that the primer pair Sp245p10 was highly specific for the Sp245 strain in the Brachiaria root and shoot field under different conditions. The qPCR assay using these primers showed differences among cultivars in the number of bacteria detected in plants after inoculation. Additionally, the number of bacteria observed in the roots was higher than that in the shoots. Conclusion: The qPCR methodology using a Sp245 strain-specific primer may be used to monitor A. baldaniorum inoculated into other plants and may find pot... Mostrar Tudo |
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Palavras-Chave: |
Azospirillum baldanioru; Plant growth promoting bacteria; Quantification; Signal grass. |
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Categoria do assunto: |
S Ciências Biológicas |
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Marc: |
LEADER 02397naa a2200277 a 4500
001 2135952
005 2021-11-09
008 2021 bl uuuu u00u1 u #d
022 $a0032-079X
024 7 $ahttps://doi.org/10.1007/s11104-021-05137-y$2DOI
100 1 $aSOARES, I. C.
245 $aReal-time PCR method to quantify Sp245 strain of Azospirillum baldaniorum on Brachiaria grasses under field conditions.$h[electronic resource]
260 $c2021
520 $aBacterial quantification by qPCR is considered the gold standard for microbial molecular diagnosis. However, a fundamental pre-requisite in this methodology is the designing of specific primers for the bacterium of interest. With the increase in bacterial genome sequencing data in the recent years, it has become possible to design specific primers that can be used to quantify different strains of the same bacterial species. Methods: To develop a real-time PCR (qPCR) protocol for the specific quantification of Azospirillum baldaniorum Sp245 strain (old Azospirillum brasilense), the Sp245 genome sequence was fragmented into small contigs with 500 base pairs each, and analyzed for similarity against the NCBI non-redundant database. A. baldaniorum-specific contigs were used to design the primers. The best pair of primers was used to quantify these bacteria after inoculation in different cultivars of Brachiaria, grown under field conditions. Results: Our results showed that the primer pair Sp245p10 was highly specific for the Sp245 strain in the Brachiaria root and shoot field under different conditions. The qPCR assay using these primers showed differences among cultivars in the number of bacteria detected in plants after inoculation. Additionally, the number of bacteria observed in the roots was higher than that in the shoots. Conclusion: The qPCR methodology using a Sp245 strain-specific primer may be used to monitor A. baldaniorum inoculated into other plants and may find potential application in field experiments.
653 $aAzospirillum baldanioru
653 $aPlant growth promoting bacteria
653 $aQuantification
653 $aSignal grass
700 1 $aPACHECO, R. S.
700 1 $aSILVA, C. G. N. da
700 1 $aSANTOS, R. S.
700 1 $aBALDANI, J. I.
700 1 $aURQUIAGA, S.
700 1 $aVIDAL, M. S.
700 1 $aARAUJO, J. L. S. de
773 $tPlant and Soil, 06 September 2021
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