Colletotrichum abscissum: Detection on symptomless citrus leaves and symptomatic citrus flowers using high-resolution melting analysis

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dc.contributor.authorGasparoto, Maria Cândida de Godoy [UNESP]
dc.contributor.authorGama, Andre Bueno
dc.contributor.authorWang, Nan-Yi
dc.contributor.authorRebello, Carolina Suguinoshita
dc.contributor.authorDewdney, Megan M.
dc.contributor.authorPeres, Natalia A.
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.date.accessioned2022-12-22T19:46:08Z
dc.date.available2022-12-22T19:46:08Z
dc.date.issued2022
dc.description.abstractHigh-resolution melting (HRM) analysis has been recently applied for the diagnosis of plant diseases. This cost-effective method is advantageous over standard PCR as it does not require DNA band visualization or the use of probes necessary for quantitative PCR. Colletotrichum abscissum causes postbloom fruit drop of citrus (PFD) and survives asymptomatically on vegetative tissue. Quiescent appressoria on citrus leaves are the primary source of inoculum between flowering periods. Early PFD symptoms may be misidentified as physical damage on citrus flowers. Our objectives were to validate an HRM-based method to rapidly identify C. abscissum on citrus leaves and flowers. We screened seven previously published primer pairs and concluded that the most effective set of primers was CaITS-F and CaITS-R815. We evaluated three different DNA extraction methods, two with a purification step (DNeasy Plant Mini and PowerSoil kits) and a faster method without a purification step (buffer GEB2). We inoculated citrus leaves with C. abscissum conidial suspensions of varying concentrations and HRM detected as few as 200 C. abscissum spores using DNeasy Plant Mini or DNeasy PowerSoil kits for DNA extraction. As expected, samples extracted with GEB2 buffer reduced HRM sensitivity. We further collected field samples from areas with high and low C. abscissum inoculum levels. The HRM method could detect the pathogen in high inoculum areas from both leaves and flowers regardless of the extraction method used. In summary, we describe a new molecular diagnostic tool to detect C. abscissum on citrus during its asymptomatic and symptomatic stages.en
dc.description.affiliationUnesp - Campus de Registro
dc.description.affiliationUniversity of Florida
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipId2019/02604-7
dc.description.versionPreprintpt
dc.identifier.doi10.1111/PPA.13688
dc.identifier.issn1365-3059
dc.identifier.lattes8048321965519076
dc.identifier.orcid0000-0001-5868-9670
dc.identifier.orcid0000-0002-0808-5294
dc.identifier.orcid0000-0001-5990-3202
dc.identifier.orcid0000-0001-7297-8030
dc.identifier.orcid0000-0003-1905-7616
dc.identifier.orcid0000-0001-5862-7963
dc.identifier.urihttp://hdl.handle.net/11449/238461
dc.language.isoeng
dc.publisherWiley
dc.relation.ispartofPlant Pathologyen
dc.rights.accessRightsAcesso aberto
dc.subjectColletotrichum acutatumen
dc.subjectAppressoriaen
dc.subjectAsymptomatic detectionen
dc.subjectColletotrichum gloeosporioidesen
dc.subjectInoculum detectionen
dc.titleColletotrichum abscissum: Detection on symptomless citrus leaves and symptomatic citrus flowers using high-resolution melting analysisen
dc.typeArtigo
unesp.campusUniversidade Estadual Paulista (Unesp), Faculdade de Ciências Agrárias do Vale do Ribeira, Registropt

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Artigos - Engenharia Agronômica - Registro