Reference gene selection for gene expression studies using RT-qPCR in virus-infected planthoppers

Autores
Maroniche, Guillermo Andrés; Sagadin, Monica Beatriz; Mongelli, Vanesa Claudia; Truol, Graciela Ana Maria; Del Vas, Mariana
Año de publicación
2011
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Background: Planthoppers not only severely affect crops by causing mechanical damage when feeding but are also vectors of several plant virus species. The analysis of gene expression in persistently infected planthoppers might unveil the molecular basis of viral transmission. Quantitative real-time RT-PCR (RT-qPCR) is currently the most accurate and sensitive method used for quantitative gene expression analysis. In order to normalize the resulting quantitative data, reference genes with constant expression during the experimental procedures are needed. Results: Partial sequences of the commonly used reference genes actin (ACT), α1-tubulin (TUB), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), elongation factor 1 alpha (EF1A), ribosomal protein S18 (RPS18) and polyubiquitin C (UBI) from Delphacodes kuscheli, a planthopper capable of persistently transmitting the plant fijivirus Mal de Río Cuarto virus (MRCV), were isolated for the first time. Specific RT-qPCR primers were designed and the expression stability of these genes was assayed in MRCV-infective and naïve planthoppers using geNorm, Normfinder and BestKeeper tools. The overall analysis showed that UBI, followed by 18S and ACT, are the most suitable genes as internal controls for quantitative gene expression studies in MRCV-infective planthoppers, while TUB and EF1A are the most variable ones. Moreover, EF1A was upregulated by MRCV infection. Conclusions: A RT-qPCR platform for gene expression analysis in the MRCV-infected planthopper vector Delphacodes kuscheli was developed. Our work is the first report on reference gene selection in virus-infected insects, and might serve as a precedent for future gene expression studies on MRCV and other virus-planthopper pathosystems.
Instituto de Biotecnología
Fil: Maroniche, Guillermo Andrés. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Sagadin, Monica Beatriz. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Fitopatología y Fisiología Vegetal (IFFIVE); Argentina
Fil: Mongelli, Vanesa Claudia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Truol, Graciela Ana Maria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Fitopatología y Fisiología Vegetal (IFFIVE); Argentina
Fil: Del Vas, Mariana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fuente
Virology Journal 8 : 308 (Diciembre 2011)
Materia
Tomato Spotted Wilt Virus
Rice Stripe Virus
Rice Black-streaked Dwarf Virus
Rice Grassy Stunt Tenuivirus
Gene Expression
Quantitative Polymerase Chain Reaction
Virus del Bronceado del Tomate
Expresión Génica
Reacción en Cadena de Polimerasa Cuantitativa
Reference Gene Selection
Selección de Genes de Referencia
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/16749
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oai_identifier_str oai:localhost:20.500.12123/16749
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spelling Reference gene selection for gene expression studies using RT-qPCR in virus-infected planthoppersMaroniche, Guillermo AndrésSagadin, Monica BeatrizMongelli, Vanesa ClaudiaTruol, Graciela Ana MariaDel Vas, MarianaTomato Spotted Wilt VirusRice Stripe VirusRice Black-streaked Dwarf VirusRice Grassy Stunt TenuivirusGene ExpressionQuantitative Polymerase Chain ReactionVirus del Bronceado del TomateExpresión GénicaReacción en Cadena de Polimerasa CuantitativaReference Gene SelectionSelección de Genes de ReferenciaBackground: Planthoppers not only severely affect crops by causing mechanical damage when feeding but are also vectors of several plant virus species. The analysis of gene expression in persistently infected planthoppers might unveil the molecular basis of viral transmission. Quantitative real-time RT-PCR (RT-qPCR) is currently the most accurate and sensitive method used for quantitative gene expression analysis. In order to normalize the resulting quantitative data, reference genes with constant expression during the experimental procedures are needed. Results: Partial sequences of the commonly used reference genes actin (ACT), α1-tubulin (TUB), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), elongation factor 1 alpha (EF1A), ribosomal protein S18 (RPS18) and polyubiquitin C (UBI) from Delphacodes kuscheli, a planthopper capable of persistently transmitting the plant fijivirus Mal de Río Cuarto virus (MRCV), were isolated for the first time. Specific RT-qPCR primers were designed and the expression stability of these genes was assayed in MRCV-infective and naïve planthoppers using geNorm, Normfinder and BestKeeper tools. The overall analysis showed that UBI, followed by 18S and ACT, are the most suitable genes as internal controls for quantitative gene expression studies in MRCV-infective planthoppers, while TUB and EF1A are the most variable ones. Moreover, EF1A was upregulated by MRCV infection. Conclusions: A RT-qPCR platform for gene expression analysis in the MRCV-infected planthopper vector Delphacodes kuscheli was developed. Our work is the first report on reference gene selection in virus-infected insects, and might serve as a precedent for future gene expression studies on MRCV and other virus-planthopper pathosystems.Instituto de BiotecnologíaFil: Maroniche, Guillermo Andrés. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaFil: Sagadin, Monica Beatriz. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Fitopatología y Fisiología Vegetal (IFFIVE); ArgentinaFil: Mongelli, Vanesa Claudia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaFil: Truol, Graciela Ana Maria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Fitopatología y Fisiología Vegetal (IFFIVE); ArgentinaFil: Del Vas, Mariana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaBioMed Central2024-02-22T11:52:22Z2024-02-22T11:52:22Z2011-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/16749https://virologyj.biomedcentral.com/articles/10.1186/1743-422X-8-3081743-422Xhttps://doi.org/10.1186/1743-422X-8-308Virology Journal 8 : 308 (Diciembre 2011)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)2025-09-04T09:50:12Zoai:localhost:20.500.12123/16749instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-09-04 09:50:12.443INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Reference gene selection for gene expression studies using RT-qPCR in virus-infected planthoppers
title Reference gene selection for gene expression studies using RT-qPCR in virus-infected planthoppers
spellingShingle Reference gene selection for gene expression studies using RT-qPCR in virus-infected planthoppers
Maroniche, Guillermo Andrés
Tomato Spotted Wilt Virus
Rice Stripe Virus
Rice Black-streaked Dwarf Virus
Rice Grassy Stunt Tenuivirus
Gene Expression
Quantitative Polymerase Chain Reaction
Virus del Bronceado del Tomate
Expresión Génica
Reacción en Cadena de Polimerasa Cuantitativa
Reference Gene Selection
Selección de Genes de Referencia
title_short Reference gene selection for gene expression studies using RT-qPCR in virus-infected planthoppers
title_full Reference gene selection for gene expression studies using RT-qPCR in virus-infected planthoppers
title_fullStr Reference gene selection for gene expression studies using RT-qPCR in virus-infected planthoppers
title_full_unstemmed Reference gene selection for gene expression studies using RT-qPCR in virus-infected planthoppers
title_sort Reference gene selection for gene expression studies using RT-qPCR in virus-infected planthoppers
dc.creator.none.fl_str_mv Maroniche, Guillermo Andrés
Sagadin, Monica Beatriz
Mongelli, Vanesa Claudia
Truol, Graciela Ana Maria
Del Vas, Mariana
author Maroniche, Guillermo Andrés
author_facet Maroniche, Guillermo Andrés
Sagadin, Monica Beatriz
Mongelli, Vanesa Claudia
Truol, Graciela Ana Maria
Del Vas, Mariana
author_role author
author2 Sagadin, Monica Beatriz
Mongelli, Vanesa Claudia
Truol, Graciela Ana Maria
Del Vas, Mariana
author2_role author
author
author
author
dc.subject.none.fl_str_mv Tomato Spotted Wilt Virus
Rice Stripe Virus
Rice Black-streaked Dwarf Virus
Rice Grassy Stunt Tenuivirus
Gene Expression
Quantitative Polymerase Chain Reaction
Virus del Bronceado del Tomate
Expresión Génica
Reacción en Cadena de Polimerasa Cuantitativa
Reference Gene Selection
Selección de Genes de Referencia
topic Tomato Spotted Wilt Virus
Rice Stripe Virus
Rice Black-streaked Dwarf Virus
Rice Grassy Stunt Tenuivirus
Gene Expression
Quantitative Polymerase Chain Reaction
Virus del Bronceado del Tomate
Expresión Génica
Reacción en Cadena de Polimerasa Cuantitativa
Reference Gene Selection
Selección de Genes de Referencia
dc.description.none.fl_txt_mv Background: Planthoppers not only severely affect crops by causing mechanical damage when feeding but are also vectors of several plant virus species. The analysis of gene expression in persistently infected planthoppers might unveil the molecular basis of viral transmission. Quantitative real-time RT-PCR (RT-qPCR) is currently the most accurate and sensitive method used for quantitative gene expression analysis. In order to normalize the resulting quantitative data, reference genes with constant expression during the experimental procedures are needed. Results: Partial sequences of the commonly used reference genes actin (ACT), α1-tubulin (TUB), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), elongation factor 1 alpha (EF1A), ribosomal protein S18 (RPS18) and polyubiquitin C (UBI) from Delphacodes kuscheli, a planthopper capable of persistently transmitting the plant fijivirus Mal de Río Cuarto virus (MRCV), were isolated for the first time. Specific RT-qPCR primers were designed and the expression stability of these genes was assayed in MRCV-infective and naïve planthoppers using geNorm, Normfinder and BestKeeper tools. The overall analysis showed that UBI, followed by 18S and ACT, are the most suitable genes as internal controls for quantitative gene expression studies in MRCV-infective planthoppers, while TUB and EF1A are the most variable ones. Moreover, EF1A was upregulated by MRCV infection. Conclusions: A RT-qPCR platform for gene expression analysis in the MRCV-infected planthopper vector Delphacodes kuscheli was developed. Our work is the first report on reference gene selection in virus-infected insects, and might serve as a precedent for future gene expression studies on MRCV and other virus-planthopper pathosystems.
Instituto de Biotecnología
Fil: Maroniche, Guillermo Andrés. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Sagadin, Monica Beatriz. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Fitopatología y Fisiología Vegetal (IFFIVE); Argentina
Fil: Mongelli, Vanesa Claudia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Truol, Graciela Ana Maria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Fitopatología y Fisiología Vegetal (IFFIVE); Argentina
Fil: Del Vas, Mariana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
description Background: Planthoppers not only severely affect crops by causing mechanical damage when feeding but are also vectors of several plant virus species. The analysis of gene expression in persistently infected planthoppers might unveil the molecular basis of viral transmission. Quantitative real-time RT-PCR (RT-qPCR) is currently the most accurate and sensitive method used for quantitative gene expression analysis. In order to normalize the resulting quantitative data, reference genes with constant expression during the experimental procedures are needed. Results: Partial sequences of the commonly used reference genes actin (ACT), α1-tubulin (TUB), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), elongation factor 1 alpha (EF1A), ribosomal protein S18 (RPS18) and polyubiquitin C (UBI) from Delphacodes kuscheli, a planthopper capable of persistently transmitting the plant fijivirus Mal de Río Cuarto virus (MRCV), were isolated for the first time. Specific RT-qPCR primers were designed and the expression stability of these genes was assayed in MRCV-infective and naïve planthoppers using geNorm, Normfinder and BestKeeper tools. The overall analysis showed that UBI, followed by 18S and ACT, are the most suitable genes as internal controls for quantitative gene expression studies in MRCV-infective planthoppers, while TUB and EF1A are the most variable ones. Moreover, EF1A was upregulated by MRCV infection. Conclusions: A RT-qPCR platform for gene expression analysis in the MRCV-infected planthopper vector Delphacodes kuscheli was developed. Our work is the first report on reference gene selection in virus-infected insects, and might serve as a precedent for future gene expression studies on MRCV and other virus-planthopper pathosystems.
publishDate 2011
dc.date.none.fl_str_mv 2011-12
2024-02-22T11:52:22Z
2024-02-22T11:52:22Z
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/20.500.12123/16749
https://virologyj.biomedcentral.com/articles/10.1186/1743-422X-8-308
1743-422X
https://doi.org/10.1186/1743-422X-8-308
url http://hdl.handle.net/20.500.12123/16749
https://virologyj.biomedcentral.com/articles/10.1186/1743-422X-8-308
https://doi.org/10.1186/1743-422X-8-308
identifier_str_mv 1743-422X
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv BioMed Central
publisher.none.fl_str_mv BioMed Central
dc.source.none.fl_str_mv Virology Journal 8 : 308 (Diciembre 2011)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
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