Selection of endogenous genes for gene expression studies in Eucalyptus under biotic (Puccinia psidii) and abiotic (acibenzolar-S-methyl) stresses using RT-qPCR

Background: Rust caused by Puccinia psidii Winter has been limiting for the establishment of new Eucalyptus plantations, as well as for resprouting of susceptible genetic materials. Identifying host genes involved in defense responses is important to elucidate resistance mechanisms. Reverse transcri...

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Autores: Boava, Leonardo P [UNESP], Laia, Marcelo L [UNESP], Jacob, Tiago R [UNESP], Dabbas, Karina M [UNESP], Gonçalves, Janaína F [UNESP], Ferro, Jesus A [UNESP], Ferro, Maria It [UNESP], Furtado, Edson L [UNESP]
Formato: artículo
Estado:Versión publicada
Fecha de publicación:2010
País:Brasil
Recursos:Universidade Estadual Paulista (UNESP)
Repositorio:Repositório Institucional da UNESP
Idioma:inglés
OAI Identifier:oai:repositorio.unesp.br:11449/71659
Acesso em linha:http://dx.doi.org/10.1186/1756-0500-3-43
http://hdl.handle.net/11449/71659
Access Level:acceso abierto
Palavra-chave:Eucalyptus
Puccinia psidii
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spelling Selection of endogenous genes for gene expression studies in Eucalyptus under biotic (Puccinia psidii) and abiotic (acibenzolar-S-methyl) stresses using RT-qPCREucalyptusPuccinia psidiiBackground: Rust caused by Puccinia psidii Winter has been limiting for the establishment of new Eucalyptus plantations, as well as for resprouting of susceptible genetic materials. Identifying host genes involved in defense responses is important to elucidate resistance mechanisms. Reverse transcription-quantitative PCR is the most common method of mRNA quantitation for gene expression analysis. This method generally employs a reference gene as an internal control to normalize results. A good endogenous control transcript shows minimal variation due to experimental conditions. Findings. We analyzed the expression of 13 genes to identify transcripts with minimal variation in leaves of 60-day-old clonal seedlings of two Eucalyptus clones (rust-resistant and susceptible) subjected to biotic (P. psidii) and abiotic (acibenzolar-S-methyl, ASM) stresses. Conclusions. For tissue samples of clones that did not receive any stimulus, a combination of the eEF2 and EglDH genes was the best control for normalization. When pathogen-inoculated and uninoculated plant samples were compared, eEF2 and UBQ together were more appropriate as normalizers. In ASM-treated and untreated leaves of both clones, transcripts of the CYP and elF4B genes combined were the ones with minimal variation. Finally, when comparing expression in both clones for ASM-treated leaves, P. psidii-inoculated leaves, ASM-treated plus P. psidii-inoculated leaves, and their respective controls, the genes with the most stable expression were EgIDH and UBQ. The chitinase gene, which is highly expressed in studies on plant resistance to phytopathogens, was used to confirm variation in gene expression due to the treatments. © 2010 Laia et al; licensee BioMed Central Ltd.Departamento de Produço Vegetal Faculdade de Cincias Agronmicas Universidade Estadual Paulista, CEP 18603-970, CP 237, Botucatu, SPDepartamento de Tecnologia Faculdade de Cincias Agrrias e Veterinrias Universidade Estadual Paulista UNESP, Via de Acesso Prof. Paulo Donato Castellane, s/n, CEP: 14884-900, Jaboticabal, SPDepartamento de Engenharia Florestal Universidade Do Estado de Santa Catarina, Av. Luiz de Cames, 2090, CEP 88520-000, Lages, Santa CatarinaDepartamento de Produço Vegetal Faculdade de Cincias Agronmicas Universidade Estadual Paulista, CEP 18603-970, CP 237, Botucatu, SPDepartamento de Tecnologia Faculdade de Cincias Agrrias e Veterinrias Universidade Estadual Paulista UNESP, Via de Acesso Prof. Paulo Donato Castellane, s/n, CEP: 14884-900, Jaboticabal, SPUniversidade Estadual Paulista (Unesp)Universidade do Estado de Santa Catarina (UDESC)2014-05-27T11:24:40Z2014-05-27T11:24:40Z2010-04-21info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://dx.doi.org/10.1186/1756-0500-3-43BMC Research Notes, v. 3.1756-0500http://hdl.handle.net/11449/7165910.1186/1756-0500-3-432-s2.0-779509259832-s2.0-77950925983.pdfScopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengBMC Research Notes0,691info:eu-repo/semantics/openAccessBoava, Leonardo P [UNESP]Laia, Marcelo L [UNESP]Jacob, Tiago R [UNESP]Dabbas, Karina M [UNESP]Gonçalves, Janaína F [UNESP]Ferro, Jesus A [UNESP]Ferro, Maria It [UNESP]Furtado, Edson L [UNESP]2025-10-22T07:15:00Zoai:repositorio.unesp.br:11449/71659Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestrepositoriounesp@unesp.bropendoar:29462025-10-22T07:15Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Selection of endogenous genes for gene expression studies in Eucalyptus under biotic (Puccinia psidii) and abiotic (acibenzolar-S-methyl) stresses using RT-qPCR
title Selection of endogenous genes for gene expression studies in Eucalyptus under biotic (Puccinia psidii) and abiotic (acibenzolar-S-methyl) stresses using RT-qPCR
spellingShingle Selection of endogenous genes for gene expression studies in Eucalyptus under biotic (Puccinia psidii) and abiotic (acibenzolar-S-methyl) stresses using RT-qPCR
Boava, Leonardo P [UNESP]
Eucalyptus
Puccinia psidii
title_short Selection of endogenous genes for gene expression studies in Eucalyptus under biotic (Puccinia psidii) and abiotic (acibenzolar-S-methyl) stresses using RT-qPCR
title_full Selection of endogenous genes for gene expression studies in Eucalyptus under biotic (Puccinia psidii) and abiotic (acibenzolar-S-methyl) stresses using RT-qPCR
title_fullStr Selection of endogenous genes for gene expression studies in Eucalyptus under biotic (Puccinia psidii) and abiotic (acibenzolar-S-methyl) stresses using RT-qPCR
title_full_unstemmed Selection of endogenous genes for gene expression studies in Eucalyptus under biotic (Puccinia psidii) and abiotic (acibenzolar-S-methyl) stresses using RT-qPCR
title_sort Selection of endogenous genes for gene expression studies in Eucalyptus under biotic (Puccinia psidii) and abiotic (acibenzolar-S-methyl) stresses using RT-qPCR
dc.creator.none.fl_str_mv Boava, Leonardo P [UNESP]
Laia, Marcelo L [UNESP]
Jacob, Tiago R [UNESP]
Dabbas, Karina M [UNESP]
Gonçalves, Janaína F [UNESP]
Ferro, Jesus A [UNESP]
Ferro, Maria It [UNESP]
Furtado, Edson L [UNESP]
author Boava, Leonardo P [UNESP]
author_facet Boava, Leonardo P [UNESP]
Laia, Marcelo L [UNESP]
Jacob, Tiago R [UNESP]
Dabbas, Karina M [UNESP]
Gonçalves, Janaína F [UNESP]
Ferro, Jesus A [UNESP]
Ferro, Maria It [UNESP]
Furtado, Edson L [UNESP]
author_role author
author2 Laia, Marcelo L [UNESP]
Jacob, Tiago R [UNESP]
Dabbas, Karina M [UNESP]
Gonçalves, Janaína F [UNESP]
Ferro, Jesus A [UNESP]
Ferro, Maria It [UNESP]
Furtado, Edson L [UNESP]
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade Estadual Paulista (Unesp)
Universidade do Estado de Santa Catarina (UDESC)
dc.subject.por.fl_str_mv Eucalyptus
Puccinia psidii
topic Eucalyptus
Puccinia psidii
description Background: Rust caused by Puccinia psidii Winter has been limiting for the establishment of new Eucalyptus plantations, as well as for resprouting of susceptible genetic materials. Identifying host genes involved in defense responses is important to elucidate resistance mechanisms. Reverse transcription-quantitative PCR is the most common method of mRNA quantitation for gene expression analysis. This method generally employs a reference gene as an internal control to normalize results. A good endogenous control transcript shows minimal variation due to experimental conditions. Findings. We analyzed the expression of 13 genes to identify transcripts with minimal variation in leaves of 60-day-old clonal seedlings of two Eucalyptus clones (rust-resistant and susceptible) subjected to biotic (P. psidii) and abiotic (acibenzolar-S-methyl, ASM) stresses. Conclusions. For tissue samples of clones that did not receive any stimulus, a combination of the eEF2 and EglDH genes was the best control for normalization. When pathogen-inoculated and uninoculated plant samples were compared, eEF2 and UBQ together were more appropriate as normalizers. In ASM-treated and untreated leaves of both clones, transcripts of the CYP and elF4B genes combined were the ones with minimal variation. Finally, when comparing expression in both clones for ASM-treated leaves, P. psidii-inoculated leaves, ASM-treated plus P. psidii-inoculated leaves, and their respective controls, the genes with the most stable expression were EgIDH and UBQ. The chitinase gene, which is highly expressed in studies on plant resistance to phytopathogens, was used to confirm variation in gene expression due to the treatments. © 2010 Laia et al; licensee BioMed Central Ltd.
publishDate 2010
dc.date.none.fl_str_mv 2010-04-21
2014-05-27T11:24:40Z
2014-05-27T11:24:40Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://dx.doi.org/10.1186/1756-0500-3-43
BMC Research Notes, v. 3.
1756-0500
http://hdl.handle.net/11449/71659
10.1186/1756-0500-3-43
2-s2.0-77950925983
2-s2.0-77950925983.pdf
url http://dx.doi.org/10.1186/1756-0500-3-43
http://hdl.handle.net/11449/71659
identifier_str_mv BMC Research Notes, v. 3.
1756-0500
10.1186/1756-0500-3-43
2-s2.0-77950925983
2-s2.0-77950925983.pdf
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv BMC Research Notes
0,691
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv Scopus
reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
repository.mail.fl_str_mv repositoriounesp@unesp.br
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