Novel set of real-time PCR primers for simultaneous detection of Liberibacter species associated with citrus Huanglongbing

Autores
Orce, Ingrid Georgina; Sendín, Lorena Noelia; Marano, Maria Rosa; Vojnov, Adrian Alberto; Castagnaro, Atilio Pedro; Filippone, María Paula
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Huanglongbing (HLB), a devastating citrus disease caused by the bacterium “Candidatus Liberibacter spp.”, is now responsible for significant economic losses worldwide. Yet, no effective disease control has been found, and the non-cultivability of the bacterium has severely hampered studies on the pathogen. The 16S rDNA gene is a well-characterized sequence, essential for cell survival, and is used for bacterial identification or assignment of close relationships at the genus and species levels. Quantitative Real-Time PCR (qPCR) assays based on 16S rDNA genes are widely used in the detection of “Ca. Liberibacter spp.” in multiplex reactions. We have developed for the first time a set of qPCR primers based on the conserved 16S rDNA gene, which specifically and simultaneously detects in a singleplex reaction, all three bacterial species associated with HLB, and can differentiateCa.Liberibacter asiaticus or africanus from americanus by their characteristic melting curves. The assay is very sensitive, and it was possible to amplify expected DNA fragments with an efficiency of 98 % using the Syber Green system and a Ct value lower than tested methods for HLB diagnosis. The application of this fast, simple and efficient detection methodology could also be important in the detection of all species of HLB-associated Liberibacters and could contribute to early pathogen detection, a crucial step in the development of preventive strategies aimed at avoiding the dissemination of this devastating disease in HLB-free areas.
Fil: Orce, Ingrid Georgina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Instituto de Tecnología Agroindustrial del Noroeste Argentino; Argentina
Fil: Sendín, Lorena Noelia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Instituto de Tecnología Agroindustrial del Noroeste Argentino; Argentina
Fil: Marano, Maria Rosa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Vojnov, Adrian Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Ciencias y Tecnología "Dr. Cesar Milstein"; Argentina
Fil: Castagnaro, Atilio Pedro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Instituto de Tecnología Agroindustrial del Noroeste Argentino; Argentina
Fil: Filippone, María Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Instituto de Tecnología Agroindustrial del Noroeste Argentino; Argentina
Materia
HUANGLONGBING
LIBERIBACTER
DIAGNOSTIC
qPCR
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/4150
Acceder
id CONICETDig_4e5d46500b9bab533d129e0cb147c688
oai_identifier_str oai:ri.conicet.gov.ar:11336/4150
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Novel set of real-time PCR primers for simultaneous detection of Liberibacter species associated with citrus HuanglongbingOrce, Ingrid GeorginaSendín, Lorena NoeliaMarano, Maria RosaVojnov, Adrian AlbertoCastagnaro, Atilio PedroFilippone, María PaulaHUANGLONGBINGLIBERIBACTERDIAGNOSTICqPCRhttps://purl.org/becyt/ford/4.1https://purl.org/becyt/ford/4Huanglongbing (HLB), a devastating citrus disease caused by the bacterium “Candidatus Liberibacter spp.”, is now responsible for significant economic losses worldwide. Yet, no effective disease control has been found, and the non-cultivability of the bacterium has severely hampered studies on the pathogen. The 16S rDNA gene is a well-characterized sequence, essential for cell survival, and is used for bacterial identification or assignment of close relationships at the genus and species levels. Quantitative Real-Time PCR (qPCR) assays based on 16S rDNA genes are widely used in the detection of “Ca. Liberibacter spp.” in multiplex reactions. We have developed for the first time a set of qPCR primers based on the conserved 16S rDNA gene, which specifically and simultaneously detects in a singleplex reaction, all three bacterial species associated with HLB, and can differentiateCa.Liberibacter asiaticus or africanus from americanus by their characteristic melting curves. The assay is very sensitive, and it was possible to amplify expected DNA fragments with an efficiency of 98 % using the Syber Green system and a Ct value lower than tested methods for HLB diagnosis. The application of this fast, simple and efficient detection methodology could also be important in the detection of all species of HLB-associated Liberibacters and could contribute to early pathogen detection, a crucial step in the development of preventive strategies aimed at avoiding the dissemination of this devastating disease in HLB-free areas.Fil: Orce, Ingrid Georgina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Instituto de Tecnología Agroindustrial del Noroeste Argentino; ArgentinaFil: Sendín, Lorena Noelia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Instituto de Tecnología Agroindustrial del Noroeste Argentino; ArgentinaFil: Marano, Maria Rosa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Vojnov, Adrian Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Ciencias y Tecnología "Dr. Cesar Milstein"; ArgentinaFil: Castagnaro, Atilio Pedro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Instituto de Tecnología Agroindustrial del Noroeste Argentino; ArgentinaFil: Filippone, María Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Instituto de Tecnología Agroindustrial del Noroeste Argentino; ArgentinaEscola Superior de Agricultura "Luiz de Queiroz"2015-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/4150Orce, Ingrid Georgina; Sendín, Lorena Noelia; Marano, Maria Rosa; Vojnov, Adrian Alberto; Castagnaro, Atilio Pedro; et al.; Novel set of real-time PCR primers for simultaneous detection of Liberibacter species associated with citrus Huanglongbing; Escola Superior de Agricultura "Luiz de Queiroz"; Scientia Agricola; 72; 3; 7-2015; 252-2590103-90161678-992Xenginfo:eu-repo/semantics/altIdentifier/url/http://www.scielo.br/scielo.php?pid=S0103-90162015000300252&script=sci_arttextinfo:eu-repo/semantics/altIdentifier/issn/0103-9016info:eu-repo/semantics/altIdentifier/doi/10.1590/0103-9016-2013-0417info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:41:53Zoai:ri.conicet.gov.ar:11336/4150instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:41:53.617CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Novel set of real-time PCR primers for simultaneous detection of Liberibacter species associated with citrus Huanglongbing
title Novel set of real-time PCR primers for simultaneous detection of Liberibacter species associated with citrus Huanglongbing
spellingShingle Novel set of real-time PCR primers for simultaneous detection of Liberibacter species associated with citrus Huanglongbing
Orce, Ingrid Georgina
HUANGLONGBING
LIBERIBACTER
DIAGNOSTIC
qPCR
title_short Novel set of real-time PCR primers for simultaneous detection of Liberibacter species associated with citrus Huanglongbing
title_full Novel set of real-time PCR primers for simultaneous detection of Liberibacter species associated with citrus Huanglongbing
title_fullStr Novel set of real-time PCR primers for simultaneous detection of Liberibacter species associated with citrus Huanglongbing
title_full_unstemmed Novel set of real-time PCR primers for simultaneous detection of Liberibacter species associated with citrus Huanglongbing
title_sort Novel set of real-time PCR primers for simultaneous detection of Liberibacter species associated with citrus Huanglongbing
dc.creator.none.fl_str_mv Orce, Ingrid Georgina
Sendín, Lorena Noelia
Marano, Maria Rosa
Vojnov, Adrian Alberto
Castagnaro, Atilio Pedro
Filippone, María Paula
author Orce, Ingrid Georgina
author_facet Orce, Ingrid Georgina
Sendín, Lorena Noelia
Marano, Maria Rosa
Vojnov, Adrian Alberto
Castagnaro, Atilio Pedro
Filippone, María Paula
author_role author
author2 Sendín, Lorena Noelia
Marano, Maria Rosa
Vojnov, Adrian Alberto
Castagnaro, Atilio Pedro
Filippone, María Paula
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv HUANGLONGBING
LIBERIBACTER
DIAGNOSTIC
qPCR
topic HUANGLONGBING
LIBERIBACTER
DIAGNOSTIC
qPCR
purl_subject.fl_str_mv https://purl.org/becyt/ford/4.1
https://purl.org/becyt/ford/4
dc.description.none.fl_txt_mv Huanglongbing (HLB), a devastating citrus disease caused by the bacterium “Candidatus Liberibacter spp.”, is now responsible for significant economic losses worldwide. Yet, no effective disease control has been found, and the non-cultivability of the bacterium has severely hampered studies on the pathogen. The 16S rDNA gene is a well-characterized sequence, essential for cell survival, and is used for bacterial identification or assignment of close relationships at the genus and species levels. Quantitative Real-Time PCR (qPCR) assays based on 16S rDNA genes are widely used in the detection of “Ca. Liberibacter spp.” in multiplex reactions. We have developed for the first time a set of qPCR primers based on the conserved 16S rDNA gene, which specifically and simultaneously detects in a singleplex reaction, all three bacterial species associated with HLB, and can differentiateCa.Liberibacter asiaticus or africanus from americanus by their characteristic melting curves. The assay is very sensitive, and it was possible to amplify expected DNA fragments with an efficiency of 98 % using the Syber Green system and a Ct value lower than tested methods for HLB diagnosis. The application of this fast, simple and efficient detection methodology could also be important in the detection of all species of HLB-associated Liberibacters and could contribute to early pathogen detection, a crucial step in the development of preventive strategies aimed at avoiding the dissemination of this devastating disease in HLB-free areas.
Fil: Orce, Ingrid Georgina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Instituto de Tecnología Agroindustrial del Noroeste Argentino; Argentina
Fil: Sendín, Lorena Noelia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Instituto de Tecnología Agroindustrial del Noroeste Argentino; Argentina
Fil: Marano, Maria Rosa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Vojnov, Adrian Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Ciencias y Tecnología "Dr. Cesar Milstein"; Argentina
Fil: Castagnaro, Atilio Pedro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Instituto de Tecnología Agroindustrial del Noroeste Argentino; Argentina
Fil: Filippone, María Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Instituto de Tecnología Agroindustrial del Noroeste Argentino; Argentina
description Huanglongbing (HLB), a devastating citrus disease caused by the bacterium “Candidatus Liberibacter spp.”, is now responsible for significant economic losses worldwide. Yet, no effective disease control has been found, and the non-cultivability of the bacterium has severely hampered studies on the pathogen. The 16S rDNA gene is a well-characterized sequence, essential for cell survival, and is used for bacterial identification or assignment of close relationships at the genus and species levels. Quantitative Real-Time PCR (qPCR) assays based on 16S rDNA genes are widely used in the detection of “Ca. Liberibacter spp.” in multiplex reactions. We have developed for the first time a set of qPCR primers based on the conserved 16S rDNA gene, which specifically and simultaneously detects in a singleplex reaction, all three bacterial species associated with HLB, and can differentiateCa.Liberibacter asiaticus or africanus from americanus by their characteristic melting curves. The assay is very sensitive, and it was possible to amplify expected DNA fragments with an efficiency of 98 % using the Syber Green system and a Ct value lower than tested methods for HLB diagnosis. The application of this fast, simple and efficient detection methodology could also be important in the detection of all species of HLB-associated Liberibacters and could contribute to early pathogen detection, a crucial step in the development of preventive strategies aimed at avoiding the dissemination of this devastating disease in HLB-free areas.
publishDate 2015
dc.date.none.fl_str_mv 2015-07
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/11336/4150
Orce, Ingrid Georgina; Sendín, Lorena Noelia; Marano, Maria Rosa; Vojnov, Adrian Alberto; Castagnaro, Atilio Pedro; et al.; Novel set of real-time PCR primers for simultaneous detection of Liberibacter species associated with citrus Huanglongbing; Escola Superior de Agricultura "Luiz de Queiroz"; Scientia Agricola; 72; 3; 7-2015; 252-259
0103-9016
1678-992X
url http://hdl.handle.net/11336/4150
identifier_str_mv Orce, Ingrid Georgina; Sendín, Lorena Noelia; Marano, Maria Rosa; Vojnov, Adrian Alberto; Castagnaro, Atilio Pedro; et al.; Novel set of real-time PCR primers for simultaneous detection of Liberibacter species associated with citrus Huanglongbing; Escola Superior de Agricultura "Luiz de Queiroz"; Scientia Agricola; 72; 3; 7-2015; 252-259
0103-9016
1678-992X
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.scielo.br/scielo.php?pid=S0103-90162015000300252&script=sci_arttext
info:eu-repo/semantics/altIdentifier/issn/0103-9016
info:eu-repo/semantics/altIdentifier/doi/10.1590/0103-9016-2013-0417
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Escola Superior de Agricultura "Luiz de Queiroz"
publisher.none.fl_str_mv Escola Superior de Agricultura "Luiz de Queiroz"
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
_version_ 1844614450658672640
score 13.070432

Publicaciones similares