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Registro Completo
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Biblioteca(s): |
Embrapa Agrobiologia. |
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Data corrente: |
24/11/2016 |
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Data da última atualização: |
23/10/2018 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Circulação/Nível: |
A - 2 |
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Autoria: |
SILVA, P. R. A. da; VIDAL, M. S.; SOARES, C. de P.; POLESE, V.; ARAUJO, J. L. S. de; BALDANI, J. I. |
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Afiliação: |
PAULA RENATA ALVES DA SILVA, UFRRJ; MARCIA SOARES VIDAL, CNPAB; CLEITON DE PAULA SOARES, BOLSISTA DA EMBRAPA AGROBIOLOGIA; VALERIA POLESE, UFRRJ; JEAN LUIZ SIMOES DE ARAUJO, CNPAB; JOSE IVO BALDANI, CNPAB. |
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Título: |
Selection and evaluation of reference genes for RT-qPCR expression studies on Burkholderia tropica strain Ppe8, a sugarcane-associated diazotrophic bacterium grown with different carbon sources or sugarcane juice |
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Ano de publicação: |
2016 |
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Fonte/Imprenta: |
Antonie van Leeuwenhoek, v. 109, n. 111, p. 1493-1502, 2016. |
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DOI: |
10.1007/s10482-016-0751-0 |
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Idioma: |
Inglês |
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Notas: |
Epub, 17 Aug., 2016. |
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Conteúdo: |
Among the members of the genus Burkholderia, Burkholderia tropica has the ability to fix nitrogen and promote sugarcane plant growth as well as act as a biological control agent. There is little information about how this bacterium metabolizes
carbohydrates as well as those carbon sources found in the sugarcane juice that accumulates in stems during plant growth. Reverse transcription quantitative PCR (RT-qPCR) can be used to evaluate changes in gene expression during bacterial growth on different carbon sources. Here we tested the expression of six reference genes, lpxC, gyrB, recA, rpoA, rpoB, and rpoD, when cells were grown with glucose, fructose, sucrose, mannitol, aconitic acid, and sugarcane juice as carbon sources. The lpxC, gyrB, and recA were selected as the most stable reference genes based on geNorm and NormFinder software analyses. Validation of these three reference genes during strain Ppe8 growth on the same carbon sources showed that genes involved in
glycogen biosynthesis (glgA, glgB, glgC) and trehalose biosynthesis (treY and treZ) were highly expressed when Ppe8 was grown in aconitic acid relative to other carbon sources, while otsA expression (trehalose biosynthesis) was reduced with all carbon
sources. In addition, the expression level of the ORF_6066 (gluconolactonase) gene was reduced on sugarcane juice. The results confirmed the stability of the three selected reference genes (lpxC, gyrB, and recA) during the RT-qPCR and also their robustness
by evaluating the relative expression of genes involved in glycogen and trehalose biosynthesis when strain Ppe8 was grown on different carbon sources and sugarcane juice. MenosAmong the members of the genus Burkholderia, Burkholderia tropica has the ability to fix nitrogen and promote sugarcane plant growth as well as act as a biological control agent. There is little information about how this bacterium metabolizes
carbohydrates as well as those carbon sources found in the sugarcane juice that accumulates in stems during plant growth. Reverse transcription quantitative PCR (RT-qPCR) can be used to evaluate changes in gene expression during bacterial growth on different carbon sources. Here we tested the expression of six reference genes, lpxC, gyrB, recA, rpoA, rpoB, and rpoD, when cells were grown with glucose, fructose, sucrose, mannitol, aconitic acid, and sugarcane juice as carbon sources. The lpxC, gyrB, and recA were selected as the most stable reference genes based on geNorm and NormFinder software analyses. Validation of these three reference genes during strain Ppe8 growth on the same carbon sources showed that genes involved in
glycogen biosynthesis (glgA, glgB, glgC) and trehalose biosynthesis (treY and treZ) were highly expressed when Ppe8 was grown in aconitic acid relative to other carbon sources, while otsA expression (trehalose biosynthesis) was reduced with all carbon
sources. In addition, the expression level of the ORF_6066 (gluconolactonase) gene was reduced on sugarcane juice. The results confirmed the stability of the three selected reference genes (lpxC, gyrB, and recA) during the RT-qPCR and also their robustness
by evalua... Mostrar Tudo |
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Palavras-Chave: |
Carbon metabolism; Diazotrophic bacteria; Normalizing genes; Real time PCR; Relative expression. |
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Categoria do assunto: |
S Ciências Biológicas |
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Marc: |
LEADER 02573naa a2200265 a 4500
001 2057134
005 2018-10-23
008 2016 bl --- 0-- u #d
024 7 $a10.1007/s10482-016-0751-0$2DOI
100 1 $aSILVA, P. R. A. da
245 $aSelection and evaluation of reference genes for RT-qPCR expression studies on Burkholderia tropica strain Ppe8, a sugarcane-associated diazotrophic bacterium grown with different carbon sources or sugarcane juice
260 $c2016
500 $aEpub, 17 Aug., 2016.
520 $aAmong the members of the genus Burkholderia, Burkholderia tropica has the ability to fix nitrogen and promote sugarcane plant growth as well as act as a biological control agent. There is little information about how this bacterium metabolizes carbohydrates as well as those carbon sources found in the sugarcane juice that accumulates in stems during plant growth. Reverse transcription quantitative PCR (RT-qPCR) can be used to evaluate changes in gene expression during bacterial growth on different carbon sources. Here we tested the expression of six reference genes, lpxC, gyrB, recA, rpoA, rpoB, and rpoD, when cells were grown with glucose, fructose, sucrose, mannitol, aconitic acid, and sugarcane juice as carbon sources. The lpxC, gyrB, and recA were selected as the most stable reference genes based on geNorm and NormFinder software analyses. Validation of these three reference genes during strain Ppe8 growth on the same carbon sources showed that genes involved in glycogen biosynthesis (glgA, glgB, glgC) and trehalose biosynthesis (treY and treZ) were highly expressed when Ppe8 was grown in aconitic acid relative to other carbon sources, while otsA expression (trehalose biosynthesis) was reduced with all carbon sources. In addition, the expression level of the ORF_6066 (gluconolactonase) gene was reduced on sugarcane juice. The results confirmed the stability of the three selected reference genes (lpxC, gyrB, and recA) during the RT-qPCR and also their robustness by evaluating the relative expression of genes involved in glycogen and trehalose biosynthesis when strain Ppe8 was grown on different carbon sources and sugarcane juice.
653 $aCarbon metabolism
653 $aDiazotrophic bacteria
653 $aNormalizing genes
653 $aReal time PCR
653 $aRelative expression
700 1 $aVIDAL, M. S.
700 1 $aSOARES, C. de P.
700 1 $aPOLESE, V.
700 1 $aARAUJO, J. L. S. de
700 1 $aBALDANI, J. I.
773 $tAntonie van Leeuwenhoek$gv. 109, n. 111, p. 1493-1502, 2016.
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