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dc.contributor.authorQuintana-González de Chaves, María
dc.contributor.authorMorán, Félix 
dc.contributor.authorBarbé, Silvia 
dc.contributor.authorBertolini, Edson
dc.contributor.authorSiverio, Felipe
dc.contributor.authorMarco-Noales, Ester 
dc.date.accessioned2023-03-03T08:45:15Z
dc.date.available2023-03-03T08:45:15Z
dc.date.issued2023es
dc.identifier.citationQuintana-González, M., Morán, F., Barbé, S., Bertolini, E., Siverio, F. & Marco-Noales, E. (2023). A new and accurate qPCR protocol to detect plant pathogenic bacteria of the genus ‘Candidatus Liberibacter’in plants and insects. Scientific Reports, 13(1), 3338.es
dc.identifier.issn2045-2322 (online)
dc.identifier.urihttps://hdl.handle.net/20.500.11939/8539
dc.description.abstractFour pathogenic bacterial species of the genus ‘Candidatus Liberibacter’, transmitted by psyllid vectors, have been associated with serious diseases affecting economically important crops of Rutaceae, Apiaceae and Solanaceae families. The most severe disease of citrus plants, huanglongbing (HLB), is associated with ‘Ca. Liberibacter asiaticus’ (CaLas), ‘Ca. Liberibacter americanus’ (CaLam) and ‘Ca. Liberibacter africanus’ (CaLaf), while ‘Ca. Liberibacter solanacearum’ (CaLsol) is associated with zebra chip disease in potatoes and vegetative disorders in apiaceous plants. Since these bacteria remain non-culturable and their symptoms are non-specific, their detection and identification are done by molecular methods, mainly based on PCR protocols. In this study, a new quantitative real-time PCR protocol based on TaqMan probe, which can also be performed in a conventional PCR version, has been developed to detect the four known phytopathogenic species of the genus Liberibacter. The new protocol has been validated according to European Plant Protection Organization (EPPO) guidelines and is able to detect CaLas, CaLam, CaLaf and CaLsol in both plants and vectors, not only using purified DNA but also using crude extracts of potato and citrus or psyllids. A comparative analysis with other previously described qPCR protocols revealed that this new one developed in this study is more specific and equally or more sensitive. Thus, other genus-specific qPCR protocols have important drawbacks regarding the lack of specificity, while with the new protocol there was no cross-reactions in 250 samples from 24 different plant and insect species from eight different geographical origins. Therefore, it can be used as a rapid and time-saving screening test, as it allows simultaneous detection of all plant pathogenic species of ‘Ca. Liberibacter’ in a one-step assay.es
dc.language.isoenes
dc.publisherSpringer Naturees
dc.rightsAttribution-NonCommercial-NoDerivatives 4.0 Internacional*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/*
dc.subjectPCR-based techniqueses
dc.subjectHuanglongbinges
dc.subjectCandidatus Liberibacter asiaticuses
dc.subjectCandidatus Liberibacter africanuses
dc.subjectCandidatus Liberibacter americanuses
dc.subjectRT-PCRes
dc.subjectDetectiones
dc.titleA new and accurate qPCR protocol to detect plant pathogenic bacteria of the genus ‘Candidatus Liberibacter’ in plants and insectses
dc.typearticlees
dc.authorAddressmarco_est@gva.eses
dc.entidadIVIACentro de Protección Vegetal y Biotecnologíaes
dc.identifier.doi10.1038/s41598-023-30345-0es
dc.identifier.urlhttps://www.nature.com/articles/s41598-023-30345-0es
dc.journal.issueNumber1es
dc.journal.titleScientific Reportses
dc.journal.volumeNumber13es
dc.page.final3338es
dc.page.initial3338es
dc.relation.projectIDThe authors would like to thank for the Grants IVIA/52202D–Sostenible–funded by Instituto Valenciano de Investigaciones Agrarias, Generalitat Valenciana, LIFE18 CCA/ES/001109 (Development of sustainable control strategies for citric under threat of climate change & preventing entry of HLB in EU -LIFE Vida for Citrus-) and PID2021-124145OR-C21 (Breeding, selection and evaluation of new citrus materials for more sustainable plantations in the face of emerging threats due to global change -FORTECITRUS-). This research was funded by INIA E-RTA2014-00008-C4-01 and INIA E-RTA2015-00005-C06-01 from the Spanish Government. MQGC is recipient of PhD grant 2017-2020 from Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria.es
dc.rights.accessRightsopenAccesses
dc.source.typeelectronicoes
dc.subject.agrisH10 Pests of plantses
dc.subject.agrisH20 Plant diseaseses
dc.subject.agrisU30 Research methodses
dc.subject.agrovocPathogens es
dc.subject.agrovocPCR es
dc.subject.agrovocPsyllids es
dc.subject.agrovocVectors es
dc.subject.agrovocCitrus es
dc.subject.agrovocprotocols es
dc.type.hasVersionpublishedVersiones


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