Development of a species-specific detection method for three Brazilian tomato begomoviruses

Detalhes bibliográficos
Autor(a) principal: Fernandes, Fernanda Rausch
Data de Publicação: 2010
Outros Autores: Albuquerque, Leonardo Cunha de, Nagata, Alice Kazuko Inoue
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UnB
Texto Completo: http://repositorio.unb.br/handle/10482/27914
https://dx.doi.org/10.1590/S1982-56762010000100007
Resumo: The begomoviruses (fam. Geminiviridae) have an ssDNA genome, are transmitted by whiteflies, and cause significant losses in tomato fields in Brazil. Nowadays, begomovirus species identification is carried out through the analysis of their complete genome sequence. Due to the high costs and difficulty in applying this technique to large-scale analysis, we aimed to develop a species-specific detection method based on PCR. Three species were targeted for amplification: Tomato severe rugose virus (ToSRV), Tomato rugose mosaic virus (ToRMV) and Tomato yellow vein streak virus (ToYVSV). Thirteen primer combinations were designed, and four were finally selected and tested against a group of 82 samples, including cloned DNA and viral DNA from infected plants previously identified by direct sequencing of PCR products. Three primer combinations were selected that could distinguish the three species, and confirmed by sequencing of amplified products. These combinations were validated by evaluation of field-collected tomato samples infected by begomovirus and this demonstrated that PCR can be a useful tool to distinguish the viruses and detect mixed infections caused by three begomovirus species.
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spelling Development of a species-specific detection method for three Brazilian tomato begomovirusesDesenvolvimento de método de detecção específico à espécie para três begomovirus no BrasilSolanum lycopersicumDetecção por PCRGeminivírusToSRVToYVSVThe begomoviruses (fam. Geminiviridae) have an ssDNA genome, are transmitted by whiteflies, and cause significant losses in tomato fields in Brazil. Nowadays, begomovirus species identification is carried out through the analysis of their complete genome sequence. Due to the high costs and difficulty in applying this technique to large-scale analysis, we aimed to develop a species-specific detection method based on PCR. Three species were targeted for amplification: Tomato severe rugose virus (ToSRV), Tomato rugose mosaic virus (ToRMV) and Tomato yellow vein streak virus (ToYVSV). Thirteen primer combinations were designed, and four were finally selected and tested against a group of 82 samples, including cloned DNA and viral DNA from infected plants previously identified by direct sequencing of PCR products. Three primer combinations were selected that could distinguish the three species, and confirmed by sequencing of amplified products. These combinations were validated by evaluation of field-collected tomato samples infected by begomovirus and this demonstrated that PCR can be a useful tool to distinguish the viruses and detect mixed infections caused by three begomovirus species.Os begomovírus (fam. Geminiviridae) possuem um genoma de ssDNA, são transmitidos por mosca-branca, e causam danos importantes em campos de produção de tomate no Brasil. Atualmente, a identificação de espécies de begomovírus é realizada pela análise da seqüência completa do genoma. Devido ao alto custo e dificuldade em se aplicar essa técnica para análises em larga escala, o objetivo desse trabalho foi desenvolver um método de detecção espécie-específico baseado em PCR. Três espécies foram avaliadas para amplificação: Tomato severe rugose virus (ToSRV), Tomato rugose mosaic virus (ToRMV) e Tomato yellow vein streak virus (ToYVSV). Foram testadas treze combinações de primers, e quatro foram selecionadas e avaliadas em um grupo de 82 amostras, incluindo DNA clonado e DNA viral total extraído de plantas infectadas com begomovírus previamente identificados por seqüenciamento direto de produto de PCR. Três combinações de primers capazes de distinguir as três espécies foram selecionadas, e confirmadas por seqüenciamento dos produtos amplificados. Estas combinações foram validadas com amostras de tomate infectadas por begomovírus em campo, demonstrando que a PCR pode ser uma ferramenta útil para separar as espécies virais e detectar infecções mistas causadas pelas três espécies de begomovírus.Em processamentoSociedade Brasileira de Fitopatologia2017-12-07T04:55:25Z2017-12-07T04:55:25Z2010info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfTrop. plant pathol.,v.35,n.1,p.043-047,2010http://repositorio.unb.br/handle/10482/27914https://dx.doi.org/10.1590/S1982-56762010000100007Fernandes, Fernanda RauschAlbuquerque, Leonardo Cunha deNagata, Alice Kazuko Inoueinfo:eu-repo/semantics/openAccessengreponame:Repositório Institucional da UnBinstname:Universidade de Brasília (UnB)instacron:UNB2024-08-28T16:17:04Zoai:repositorio.unb.br:10482/27914Repositório InstitucionalPUBhttps://repositorio.unb.br/oai/requestrepositorio@unb.bropendoar:2024-08-28T16:17:04Repositório Institucional da UnB - Universidade de Brasília (UnB)false
dc.title.none.fl_str_mv Development of a species-specific detection method for three Brazilian tomato begomoviruses
Desenvolvimento de método de detecção específico à espécie para três begomovirus no Brasil
title Development of a species-specific detection method for three Brazilian tomato begomoviruses
spellingShingle Development of a species-specific detection method for three Brazilian tomato begomoviruses
Fernandes, Fernanda Rausch
Solanum lycopersicum
Detecção por PCR
Geminivírus
ToSRV
ToYVSV
title_short Development of a species-specific detection method for three Brazilian tomato begomoviruses
title_full Development of a species-specific detection method for three Brazilian tomato begomoviruses
title_fullStr Development of a species-specific detection method for three Brazilian tomato begomoviruses
title_full_unstemmed Development of a species-specific detection method for three Brazilian tomato begomoviruses
title_sort Development of a species-specific detection method for three Brazilian tomato begomoviruses
author Fernandes, Fernanda Rausch
author_facet Fernandes, Fernanda Rausch
Albuquerque, Leonardo Cunha de
Nagata, Alice Kazuko Inoue
author_role author
author2 Albuquerque, Leonardo Cunha de
Nagata, Alice Kazuko Inoue
author2_role author
author
dc.contributor.author.fl_str_mv Fernandes, Fernanda Rausch
Albuquerque, Leonardo Cunha de
Nagata, Alice Kazuko Inoue
dc.subject.por.fl_str_mv Solanum lycopersicum
Detecção por PCR
Geminivírus
ToSRV
ToYVSV
topic Solanum lycopersicum
Detecção por PCR
Geminivírus
ToSRV
ToYVSV
description The begomoviruses (fam. Geminiviridae) have an ssDNA genome, are transmitted by whiteflies, and cause significant losses in tomato fields in Brazil. Nowadays, begomovirus species identification is carried out through the analysis of their complete genome sequence. Due to the high costs and difficulty in applying this technique to large-scale analysis, we aimed to develop a species-specific detection method based on PCR. Three species were targeted for amplification: Tomato severe rugose virus (ToSRV), Tomato rugose mosaic virus (ToRMV) and Tomato yellow vein streak virus (ToYVSV). Thirteen primer combinations were designed, and four were finally selected and tested against a group of 82 samples, including cloned DNA and viral DNA from infected plants previously identified by direct sequencing of PCR products. Three primer combinations were selected that could distinguish the three species, and confirmed by sequencing of amplified products. These combinations were validated by evaluation of field-collected tomato samples infected by begomovirus and this demonstrated that PCR can be a useful tool to distinguish the viruses and detect mixed infections caused by three begomovirus species.
publishDate 2010
dc.date.none.fl_str_mv 2010
2017-12-07T04:55:25Z
2017-12-07T04:55:25Z
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 Trop. plant pathol.,v.35,n.1,p.043-047,2010
http://repositorio.unb.br/handle/10482/27914
https://dx.doi.org/10.1590/S1982-56762010000100007
identifier_str_mv Trop. plant pathol.,v.35,n.1,p.043-047,2010
url http://repositorio.unb.br/handle/10482/27914
https://dx.doi.org/10.1590/S1982-56762010000100007
dc.language.iso.fl_str_mv eng
language eng
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.publisher.none.fl_str_mv Sociedade Brasileira de Fitopatologia
publisher.none.fl_str_mv Sociedade Brasileira de Fitopatologia
dc.source.none.fl_str_mv reponame:Repositório Institucional da UnB
instname:Universidade de Brasília (UnB)
instacron:UNB
instname_str Universidade de Brasília (UnB)
instacron_str UNB
institution UNB
reponame_str Repositório Institucional da UnB
collection Repositório Institucional da UnB
repository.name.fl_str_mv Repositório Institucional da UnB - Universidade de Brasília (UnB)
repository.mail.fl_str_mv repositorio@unb.br
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