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 | Acesso ao texto completo restrito à biblioteca da Embrapa Pecuária Sudeste. Para informações adicionais entre em contato com cppse.biblioteca@embrapa.br. |
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Registro Completo |
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Biblioteca(s): |
Embrapa Pecuária Sudeste. |
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Data corrente: |
28/08/2018 |
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Data da última atualização: |
12/09/2018 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Autoria: |
TOSCANO, J. H. B.; LOPES, L. G.; GIRALDELO, L. A.; SILVA, M. H. da; OKINO, C. H.; CHAGAS, A. C. de S. |
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Afiliação: |
João Henrique Barbosa Toscano, UNESP; Louyse Gabrielli Lopes, UNICEP; Luciana Aparecida Giraldelo, UNICEP; Matheus Henrique da Silva, UNICEP; CINTIA HIROMI OKINO, CPPSE; ANA CAROLINA DE SOUZA CHAGAS, CPPSE. |
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Título: |
Identification of appropriate reference genes for local immune-related studies in Morada Nova sheep infected with Haemonchus contortus. |
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Ano de publicação: |
2018 |
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Fonte/Imprenta: |
Molecular Biology Reports, v.45, n.5, p.1253-1262, out. 2018. |
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Idioma: |
Inglês |
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Conteúdo: |
Due to the great economic impact of Haemonchus contortus on sheep farming, there is an increasing number of studies addressing host resistance against this nematode, including identification of directly related immune mechanisms. In this context, relative gene expression by RT-qPCR have been largely used, due to its rapidity, high sensitivity, specificity, and reproducibility. Although, appropriate reference gene selection is crucial for accurate interpretation of results. In this study, five reference genes (GAPDH, G6PDH, YWHAZ, ACTB, and B2M) were tested for expression stability in abomasum (fundic and pyloric regions) and abomasal lymph nodes of Morada Nova sheep classified as resistant (n = 5) or susceptible (n = 5) to H. contortus infection in a flock of 151 animals. GAPDH combined with YWHAZ were selected as reference genes for abomasal fundic region and abomasal lymph nodes, whereas YWHAZ was the most stable gene for abomasal pyloric region. These genes presented the lowest intra- and inter-group variations and, consequently, highest stability. In contrast, expression of G6PDH was the least stable in all tissues. The impact of reference gene selection was demonstrated by relative quantification of a pro-inflammatory cytokine (TNFα) in abomasal fundic region. Significant differences in TNFα expression levels between resistant and susceptible groups were only observed when the most stable genes (GAPDH combined with YWHAZ) or GAPDH were used as reference genes, whereas no significant differences were observed when other tested reference genes were used. It was demonstrated that normalization of expression data using inappropriate reference genes may significantly influence interpretation results. MenosDue to the great economic impact of Haemonchus contortus on sheep farming, there is an increasing number of studies addressing host resistance against this nematode, including identification of directly related immune mechanisms. In this context, relative gene expression by RT-qPCR have been largely used, due to its rapidity, high sensitivity, specificity, and reproducibility. Although, appropriate reference gene selection is crucial for accurate interpretation of results. In this study, five reference genes (GAPDH, G6PDH, YWHAZ, ACTB, and B2M) were tested for expression stability in abomasum (fundic and pyloric regions) and abomasal lymph nodes of Morada Nova sheep classified as resistant (n = 5) or susceptible (n = 5) to H. contortus infection in a flock of 151 animals. GAPDH combined with YWHAZ were selected as reference genes for abomasal fundic region and abomasal lymph nodes, whereas YWHAZ was the most stable gene for abomasal pyloric region. These genes presented the lowest intra- and inter-group variations and, consequently, highest stability. In contrast, expression of G6PDH was the least stable in all tissues. The impact of reference gene selection was demonstrated by relative quantification of a pro-inflammatory cytokine (TNFα) in abomasal fundic region. Significant differences in TNFα expression levels between resistant and susceptible groups were only observed when the most stable genes (GAPDH combined with YWHAZ) or GAPDH were used as reference genes,... Mostrar Tudo |
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Palavras-Chave: |
Genes de referência. |
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Thesagro: |
Haemonchus Contortus; Ovino. |
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Thesaurus Nal: |
Sheep. |
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Categoria do assunto: |
G Melhoramento Genético |
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Marc: |
LEADER 02463naa a2200229 a 4500
001 2094611
005 2018-09-12
008 2018 bl uuuu u00u1 u #d
100 1 $aTOSCANO, J. H. B.
245 $aIdentification of appropriate reference genes for local immune-related studies in Morada Nova sheep infected with Haemonchus contortus.$h[electronic resource]
260 $c2018
520 $aDue to the great economic impact of Haemonchus contortus on sheep farming, there is an increasing number of studies addressing host resistance against this nematode, including identification of directly related immune mechanisms. In this context, relative gene expression by RT-qPCR have been largely used, due to its rapidity, high sensitivity, specificity, and reproducibility. Although, appropriate reference gene selection is crucial for accurate interpretation of results. In this study, five reference genes (GAPDH, G6PDH, YWHAZ, ACTB, and B2M) were tested for expression stability in abomasum (fundic and pyloric regions) and abomasal lymph nodes of Morada Nova sheep classified as resistant (n = 5) or susceptible (n = 5) to H. contortus infection in a flock of 151 animals. GAPDH combined with YWHAZ were selected as reference genes for abomasal fundic region and abomasal lymph nodes, whereas YWHAZ was the most stable gene for abomasal pyloric region. These genes presented the lowest intra- and inter-group variations and, consequently, highest stability. In contrast, expression of G6PDH was the least stable in all tissues. The impact of reference gene selection was demonstrated by relative quantification of a pro-inflammatory cytokine (TNFα) in abomasal fundic region. Significant differences in TNFα expression levels between resistant and susceptible groups were only observed when the most stable genes (GAPDH combined with YWHAZ) or GAPDH were used as reference genes, whereas no significant differences were observed when other tested reference genes were used. It was demonstrated that normalization of expression data using inappropriate reference genes may significantly influence interpretation results.
650 $aSheep
650 $aHaemonchus Contortus
650 $aOvino
653 $aGenes de referência
700 1 $aLOPES, L. G.
700 1 $aGIRALDELO, L. A.
700 1 $aSILVA, M. H. da
700 1 $aOKINO, C. H.
700 1 $aCHAGAS, A. C. de S.
773 $tMolecular Biology Reports$gv.45, n.5, p.1253-1262, out. 2018.
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Registro original: |
Embrapa Pecuária Sudeste (CPPSE) |
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| Acesso ao texto completo restrito à biblioteca da Embrapa Meio Ambiente. Para informações adicionais entre em contato com cnpma.biblioteca@embrapa.br. |
Registro Completo
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Biblioteca(s): |
Embrapa Meio Ambiente. |
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Data corrente: |
08/02/2018 |
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Data da última atualização: |
08/02/2018 |
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Tipo da produção científica: |
Resumo em Anais de Congresso |
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Autoria: |
MENDES, L. W.; RAAIJMAKERS, J. M.; HOLLANDER, M. de; MENDES, R.; TSAI, S. M. |
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Afiliação: |
L. W. MENDES; J. M. RAAIJMAKERS; M. de HOLLANDER; RODRIGO MENDES, CNPMA; S. M. TSAI. |
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Título: |
Impact of resistance breeding on rhizosphere microbiome assembly in common bean. |
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Ano de publicação: |
2017 |
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Fonte/Imprenta: |
In: SYMPOSIUM ON BACTERIAL GENETICS AND ECOLOGY, 14., 2017, Aberdeen. Annals... Aberdeen: BAGECO, 2017. |
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Páginas: |
p. 105 |
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Idioma: |
Inglês |
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Conteúdo: |
The rhizosphere microbiome plays a key role in plant growth and health, providing a first line of defense against pathogen root infection. Here, we investigated how breeding for resistance of common bean (Phaseolus vulgaris) to the soil borne pathogen Fusarium oxysporum (Fox) changes the composition and metabolic potential of the bacterial community in the rhizosphere. For different bean cultivars grown in two contrasting soils, rhizobacterial abundance was positively correlated with Fox-resistance. Pseudomonadaceae, Bacillaceae, Solibacteraceae and Cytophagaceae are more abundant in the rhizosphere of the resistant cultivar. Network analyses showed a non modular topology of the rhizosphere microbiome of the Fox-resistant cultivar, suggesting a more complex and highly connected bacterial community when compared to the Fox-susceptible cultivar. |
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Thesagro: |
Feijão; Rizosfera. |
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Categoria do assunto: |
V Taxonomia de Organismos |
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Marc: |
LEADER 01481naa a2200205 a 4500
001 2087509
005 2018-02-08
008 2017 bl --- 0-- u #d
100 1 $aMENDES, L. W.
245 $aImpact of resistance breeding on rhizosphere microbiome assembly in common bean.$h[electronic resource]
260 $c2017
300 $ap. 105
520 $aThe rhizosphere microbiome plays a key role in plant growth and health, providing a first line of defense against pathogen root infection. Here, we investigated how breeding for resistance of common bean (Phaseolus vulgaris) to the soil borne pathogen Fusarium oxysporum (Fox) changes the composition and metabolic potential of the bacterial community in the rhizosphere. For different bean cultivars grown in two contrasting soils, rhizobacterial abundance was positively correlated with Fox-resistance. Pseudomonadaceae, Bacillaceae, Solibacteraceae and Cytophagaceae are more abundant in the rhizosphere of the resistant cultivar. Network analyses showed a non modular topology of the rhizosphere microbiome of the Fox-resistant cultivar, suggesting a more complex and highly connected bacterial community when compared to the Fox-susceptible cultivar.
650 $aFeijão
650 $aRizosfera
700 1 $aRAAIJMAKERS, J. M.
700 1 $aHOLLANDER, M. de
700 1 $aMENDES, R.
700 1 $aTSAI, S. M.
773 $tIn: SYMPOSIUM ON BACTERIAL GENETICS AND ECOLOGY, 14., 2017, Aberdeen. Annals... Aberdeen: BAGECO, 2017.
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