Colletotrichum abscissum: Detection on symptomless citrus leaves and symptomatic citrus flowers using high-resolution melting analysis
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://hdl.handle.net/11449/238461 |
Resumo: | High-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. |
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Colletotrichum abscissum: Detection on symptomless citrus leaves and symptomatic citrus flowers using high-resolution melting analysisColletotrichum acutatumAppressoriaAsymptomatic detectionColletotrichum gloeosporioidesInoculum detectionHigh-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.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)PreprintUnesp - Campus de RegistroUniversity of Florida2019/02604-7WileyUniversidade Estadual Paulista (Unesp)Gasparoto, Maria Cândida de Godoy [UNESP]Gama, Andre BuenoWang, Nan-YiRebello, Carolina SuguinoshitaDewdney, Megan M.Peres, Natalia A.2022-12-22T19:46:08Z2022-12-22T19:46:08Z2022info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdf1365-3059http://hdl.handle.net/11449/23846110.1111/PPA.1368880483219655190760000-0001-5868-96700000-0002-0808-52940000-0001-5990-32020000-0001-7297-80300000-0003-1905-76160000-0001-5862-7963engPlant Pathologyinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESP2024-05-03T13:20:08Zoai:repositorio.unesp.br:11449/238461Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-05-03T13:20:08Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Colletotrichum abscissum: Detection on symptomless citrus leaves and symptomatic citrus flowers using high-resolution melting analysis |
title |
Colletotrichum abscissum: Detection on symptomless citrus leaves and symptomatic citrus flowers using high-resolution melting analysis |
spellingShingle |
Colletotrichum abscissum: Detection on symptomless citrus leaves and symptomatic citrus flowers using high-resolution melting analysis Gasparoto, Maria Cândida de Godoy [UNESP] Colletotrichum acutatum Appressoria Asymptomatic detection Colletotrichum gloeosporioides Inoculum detection |
title_short |
Colletotrichum abscissum: Detection on symptomless citrus leaves and symptomatic citrus flowers using high-resolution melting analysis |
title_full |
Colletotrichum abscissum: Detection on symptomless citrus leaves and symptomatic citrus flowers using high-resolution melting analysis |
title_fullStr |
Colletotrichum abscissum: Detection on symptomless citrus leaves and symptomatic citrus flowers using high-resolution melting analysis |
title_full_unstemmed |
Colletotrichum abscissum: Detection on symptomless citrus leaves and symptomatic citrus flowers using high-resolution melting analysis |
title_sort |
Colletotrichum abscissum: Detection on symptomless citrus leaves and symptomatic citrus flowers using high-resolution melting analysis |
author |
Gasparoto, Maria Cândida de Godoy [UNESP] |
author_facet |
Gasparoto, Maria Cândida de Godoy [UNESP] Gama, Andre Bueno Wang, Nan-Yi Rebello, Carolina Suguinoshita Dewdney, Megan M. Peres, Natalia A. |
author_role |
author |
author2 |
Gama, Andre Bueno Wang, Nan-Yi Rebello, Carolina Suguinoshita Dewdney, Megan M. Peres, Natalia A. |
author2_role |
author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
Gasparoto, Maria Cândida de Godoy [UNESP] Gama, Andre Bueno Wang, Nan-Yi Rebello, Carolina Suguinoshita Dewdney, Megan M. Peres, Natalia A. |
dc.subject.por.fl_str_mv |
Colletotrichum acutatum Appressoria Asymptomatic detection Colletotrichum gloeosporioides Inoculum detection |
topic |
Colletotrichum acutatum Appressoria Asymptomatic detection Colletotrichum gloeosporioides Inoculum detection |
description |
High-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. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-12-22T19:46:08Z 2022-12-22T19:46:08Z 2022 |
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 |
1365-3059 http://hdl.handle.net/11449/238461 10.1111/PPA.13688 8048321965519076 0000-0001-5868-9670 0000-0002-0808-5294 0000-0001-5990-3202 0000-0001-7297-8030 0000-0003-1905-7616 0000-0001-5862-7963 |
identifier_str_mv |
1365-3059 10.1111/PPA.13688 8048321965519076 0000-0001-5868-9670 0000-0002-0808-5294 0000-0001-5990-3202 0000-0001-7297-8030 0000-0003-1905-7616 0000-0001-5862-7963 |
url |
http://hdl.handle.net/11449/238461 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Plant Pathology |
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 |
Wiley |
publisher.none.fl_str_mv |
Wiley |
dc.source.none.fl_str_mv |
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_ |
1799964910433599488 |