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CRISPR genotyping as complementary tool for epidemiological surveillance of Erwinia amylovora outbreaks

2021Journal article Open access
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dc.contributor.authorMendes, Rafael
dc.contributor.authorLuz, J.P.
dc.contributor.authorSantos, Conceição
dc.contributor.authorTavares, Fernando
dc.date.accessioned2022-01-04T16:58:50Z
dc.date.available2022-01-04T16:58:50Z
dc.date.issued2021
dc.description.abstractFire blight is a destructive plant disease caused by Erwinia amylovora affecting pome fruit trees, and responsible for large yield declines, long phytosanitary confinements, and high economic losses. In Portugal, the first major fire blight outbreaks occurred in 2010 and 2011, and although later considered eradicated, the emergence of other outbreaks in recent years stressed the need to characterize the E. amylovora populations associated with these outbreaks. In this regard, CRISPR genotyping, assessment of three virulence markers, and semi-quantitative virulence bioassays, were carried out to determine the genotype, and assess the virulence of thirty-six E. amylovora isolates associated with outbreaks occurring between 2010 and 2017 and affecting apple and pear orchards located in the country central-west, known as the main producing region of pome fruits in Portugal. The data gathered reveal that 35 E. amylovora isolates belong to one of the widely-distributed CRISPR genotypes (5-24-38 / D-a-α) regardless the host species, year and region. Ea 680 was the single isolate revealing a new CRISPR genotype due to a novel CR2 spacer located closer to the leader sequence and therefore thought to be recently acquired. Regarding pathogenicity, although dot-blot hybridization assays showed the presence of key virulence factors, namely hrpL (T3SS), hrpN (T3E) and amsG from the amylovoran biosynthesis operon in all E. amylovora isolates studied, pathogenicity bioassays on immature pear slices allowed to distinguish four virulence levels, with most of the isolates revealing an intermediate to severe virulence phenotype. Regardless the clonal population structure of the E. amylovora associated to the outbreaks occurring in Portugal between 2010 and 2017, the different virulence phenotypes, suggests that E. amylovora may have been introduced at different instances into the country. This is the first study regarding E. amylovora in Portugal, and it discloses a novel CRISPR genotype for this bacterium.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationMendes RJ, Luz JP, Santos C, Tavares F (2021) CRISPR genotyping as complementary tool for epidemiological surveillance of Erwinia amylovora outbreaks. PLoS ONE 16(4): e0250280pt_PT
dc.identifier.doi10.1371/journal.pone.0250280pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.11/7787
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.relationRM received support for this study from the Fundação para a Ciência e Tecnologia, grant number SFRH/BD/133519/2017, [https://www.fct.pt/index.phtml.en]pt_PT
dc.relationFT received support from the European Union's Horizon 2020 Research and Innovation Programme, grant number 857251, [https://ec.europa.eu/programmes/horizon2020/en].pt_PT
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.subjectErwinia amylovorapt_PT
dc.subjectGenotypingpt_PT
dc.titleCRISPR genotyping as complementary tool for epidemiological surveillance of Erwinia amylovora outbreakspt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.issue4pt_PT
oaire.citation.startPagee0250280pt_PT
oaire.citation.titlePLOS ONEpt_PT
oaire.citation.volume16pt_PT
person.familyNameMendes
person.familyNameLuz
person.familyNameDos Santos
person.familyNameTavares
person.givenNameRafael
person.givenNameJoão Pedro
person.givenNameConceição Domingos José
person.givenNameFernando
person.identifier.ciencia-idA712-701F-0EEC
person.identifier.ciencia-idCF19-F43F-86DE
person.identifier.ciencia-idA61A-A0F5-77F8
person.identifier.orcid0000-0001-5212-6172
person.identifier.orcid0000-0002-3494-737X
person.identifier.orcid0000-0001-9913-1155
person.identifier.ridM-7245-2013
person.identifier.ridC-3375-2013
person.identifier.scopus-author-id57191629737
person.identifier.scopus-author-id36504399900
person.identifier.scopus-author-id7005803189
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
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relation.isAuthorOfPublication.latestForDiscovery7cb85c28-f773-48e3-9e1c-6ffd0f310aef

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