A nested-PCR assay for detection of Xylella fastidiosa in citrus plants and sharpshooter leafhoppers
Autor(a) principal: | |
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Data de Publicação: | 2004 |
Outros Autores: | , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1111/j.1365-2672.2004.02176.x http://hdl.handle.net/11449/3927 |
Resumo: | Aims: Detection of Xylella fastidiosa in citrus plants and insect vectors.Methods and Results: Chelex 100 resin matrix was successfully standardized allowing a fast DNA extraction of X. fastidiosa. An amplicon of 500 bp was observed in samples of citrus leaf and citrus xylem extract, with and without symptoms of citrus variegated chlorosis, using PCR with a specific primer set indicating the presence of X. fastidiosa. The addition of insoluble acid-washed polyvinylpyrrolidone (PVPP) prior to DNA extraction of insect samples using Chelex 100 resin together with nested-PCR permitted the detection of X. fastidiosa within sharpshooter heads with great sensitivity. It was possible to detect up to two bacteria per reaction. From 250 sharpshooter samples comprising four species (Dilobopterus costalimai, Oncometopia facialis, Bucephalogonia xanthopis and Acrogonia sp.), 87 individuals showed positive results for X. fastidiosa in a nested-PCR assay.Conclusions: the use of Chelex 100 resin allowed a fast and efficient DNA extraction to be used in the detection of X. fastidiosa in citrus plants and insect vectors by PCR and nested-PCR assays, respectively.Significance and Impact of the study: the employment of efficient and sensitive methods to detect X. fastidiosa in citrus plants and insect vectors will greatly assist epidemiological studies. |
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A nested-PCR assay for detection of Xylella fastidiosa in citrus plants and sharpshooter leafhoppersChelex 100citrus variegated chlorosisDNA extractionNested PCRsharpshooterXylella fastidiosaAims: Detection of Xylella fastidiosa in citrus plants and insect vectors.Methods and Results: Chelex 100 resin matrix was successfully standardized allowing a fast DNA extraction of X. fastidiosa. An amplicon of 500 bp was observed in samples of citrus leaf and citrus xylem extract, with and without symptoms of citrus variegated chlorosis, using PCR with a specific primer set indicating the presence of X. fastidiosa. The addition of insoluble acid-washed polyvinylpyrrolidone (PVPP) prior to DNA extraction of insect samples using Chelex 100 resin together with nested-PCR permitted the detection of X. fastidiosa within sharpshooter heads with great sensitivity. It was possible to detect up to two bacteria per reaction. From 250 sharpshooter samples comprising four species (Dilobopterus costalimai, Oncometopia facialis, Bucephalogonia xanthopis and Acrogonia sp.), 87 individuals showed positive results for X. fastidiosa in a nested-PCR assay.Conclusions: the use of Chelex 100 resin allowed a fast and efficient DNA extraction to be used in the detection of X. fastidiosa in citrus plants and insect vectors by PCR and nested-PCR assays, respectively.Significance and Impact of the study: the employment of efficient and sensitive methods to detect X. fastidiosa in citrus plants and insect vectors will greatly assist epidemiological studies.Univ Estadual Paulista, Fac Ciências Agr & Vet, Dept Technol, Lab Bioquim Microrganismos & Plantas, Jaboticabal, SP, BrazilUniv Estadual Paulista, Fac Ciências Agr & Vet, Dept Technol, Lab Bioquim Microrganismos & Plantas, Jaboticabal, SP, BrazilBlackwell PublishingUniversidade Estadual Paulista (Unesp)Ciapina, L. P.Alves, LMCLemos, EGM2014-05-20T13:17:29Z2014-05-20T13:17:29Z2004-01-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article546-551application/pdfhttp://dx.doi.org/10.1111/j.1365-2672.2004.02176.xJournal of Applied Microbiology. Oxford: Blackwell Publishing Ltd, v. 96, n. 3, p. 546-551, 2004.1364-5072http://hdl.handle.net/11449/392710.1111/j.1365-2672.2004.02176.xWOS:000188990400014WOS000188990400014.pdf6676176632132637Web of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengJournal of Applied Microbiology2.1600,795info:eu-repo/semantics/openAccess2024-06-07T15:31:35Zoai:repositorio.unesp.br:11449/3927Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T16:13:38.242387Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
A nested-PCR assay for detection of Xylella fastidiosa in citrus plants and sharpshooter leafhoppers |
title |
A nested-PCR assay for detection of Xylella fastidiosa in citrus plants and sharpshooter leafhoppers |
spellingShingle |
A nested-PCR assay for detection of Xylella fastidiosa in citrus plants and sharpshooter leafhoppers Ciapina, L. P. Chelex 100 citrus variegated chlorosis DNA extraction Nested PCR sharpshooter Xylella fastidiosa |
title_short |
A nested-PCR assay for detection of Xylella fastidiosa in citrus plants and sharpshooter leafhoppers |
title_full |
A nested-PCR assay for detection of Xylella fastidiosa in citrus plants and sharpshooter leafhoppers |
title_fullStr |
A nested-PCR assay for detection of Xylella fastidiosa in citrus plants and sharpshooter leafhoppers |
title_full_unstemmed |
A nested-PCR assay for detection of Xylella fastidiosa in citrus plants and sharpshooter leafhoppers |
title_sort |
A nested-PCR assay for detection of Xylella fastidiosa in citrus plants and sharpshooter leafhoppers |
author |
Ciapina, L. P. |
author_facet |
Ciapina, L. P. Alves, LMC Lemos, EGM |
author_role |
author |
author2 |
Alves, LMC Lemos, EGM |
author2_role |
author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
Ciapina, L. P. Alves, LMC Lemos, EGM |
dc.subject.por.fl_str_mv |
Chelex 100 citrus variegated chlorosis DNA extraction Nested PCR sharpshooter Xylella fastidiosa |
topic |
Chelex 100 citrus variegated chlorosis DNA extraction Nested PCR sharpshooter Xylella fastidiosa |
description |
Aims: Detection of Xylella fastidiosa in citrus plants and insect vectors.Methods and Results: Chelex 100 resin matrix was successfully standardized allowing a fast DNA extraction of X. fastidiosa. An amplicon of 500 bp was observed in samples of citrus leaf and citrus xylem extract, with and without symptoms of citrus variegated chlorosis, using PCR with a specific primer set indicating the presence of X. fastidiosa. The addition of insoluble acid-washed polyvinylpyrrolidone (PVPP) prior to DNA extraction of insect samples using Chelex 100 resin together with nested-PCR permitted the detection of X. fastidiosa within sharpshooter heads with great sensitivity. It was possible to detect up to two bacteria per reaction. From 250 sharpshooter samples comprising four species (Dilobopterus costalimai, Oncometopia facialis, Bucephalogonia xanthopis and Acrogonia sp.), 87 individuals showed positive results for X. fastidiosa in a nested-PCR assay.Conclusions: the use of Chelex 100 resin allowed a fast and efficient DNA extraction to be used in the detection of X. fastidiosa in citrus plants and insect vectors by PCR and nested-PCR assays, respectively.Significance and Impact of the study: the employment of efficient and sensitive methods to detect X. fastidiosa in citrus plants and insect vectors will greatly assist epidemiological studies. |
publishDate |
2004 |
dc.date.none.fl_str_mv |
2004-01-01 2014-05-20T13:17:29Z 2014-05-20T13:17:29Z |
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/10.1111/j.1365-2672.2004.02176.x Journal of Applied Microbiology. Oxford: Blackwell Publishing Ltd, v. 96, n. 3, p. 546-551, 2004. 1364-5072 http://hdl.handle.net/11449/3927 10.1111/j.1365-2672.2004.02176.x WOS:000188990400014 WOS000188990400014.pdf 6676176632132637 |
url |
http://dx.doi.org/10.1111/j.1365-2672.2004.02176.x http://hdl.handle.net/11449/3927 |
identifier_str_mv |
Journal of Applied Microbiology. Oxford: Blackwell Publishing Ltd, v. 96, n. 3, p. 546-551, 2004. 1364-5072 10.1111/j.1365-2672.2004.02176.x WOS:000188990400014 WOS000188990400014.pdf 6676176632132637 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Journal of Applied Microbiology 2.160 0,795 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
546-551 application/pdf |
dc.publisher.none.fl_str_mv |
Blackwell Publishing |
publisher.none.fl_str_mv |
Blackwell Publishing |
dc.source.none.fl_str_mv |
Web of Science 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 |
|
_version_ |
1808128620829343744 |