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Cloning, characterization, in vitro and in planta expression of a necrosis-inducing Phytophthora protein 1 gene npp1 from Phytophthora cinnamomi

2019Journal article Restricted access
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dc.contributor.authorMartins, Ivone M.
dc.contributor.authorMeirinho, Sofia
dc.contributor.authorCosta, Rodrigo
dc.contributor.authorCravador, Alfredo
dc.contributor.authorChoupina, Altino
dc.date.accessioned2020-02-06T15:23:26Z
dc.date.available2020-02-06T15:23:26Z
dc.date.issued2019
dc.description.abstractThe soil-borne oomycete Phytophthora cinnamomi is a highly destructive Phytophthora species associated with the decline of forest. This pathogen secretes a novel class of necrosis-inducing proteins known as Nep1-like proteins (NLPs). In this work, we report the sequencing and molecular characterization of one of these proteins, more specifically the necrosis-inducing Phytophthora protein 1 (NPP1). The ORF of the npp1 gene (EMBL database AM403130) has 768 bp encoding a putative peptide of 256 amino acids with a molecular weight of approximately 25 kD. In order to understand its function, in vitro gene expression was studied during growth in different carbon sources (glucose, cellulose, and sawdust), and at different times of infection, in vivo by RT-qPCR. The highest expression of the npp1 gene occurred in glucose medium followed by sawdust. In vivo infection of Castanea sativa roots with P. cinnamomi revealed a decrease in npp1 expression from 12 to 24 h; at 36 h its expression increased suggesting the existence of a complex mechanism of defense/attack interaction between the pathogen and the host. Expression of recombinant npp1 gene was achieved in Pichia pastoris and assessed by SDS-PAGE analysis of the protein secreted into the culture supernatant, revealing the presence of the NPP1 protein.pt_PT
dc.description.sponsorshipFunding Agency European Union (EU); COMBATINTA/SP2; P11/02pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.doi10.1007/s11033-019-05091-0pt_PT
dc.identifier.issn0301-4851
dc.identifier.urihttp://hdl.handle.net/10400.1/13467
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherSpringer Naturept_PT
dc.subjectCastanea sativapt_PT
dc.subjectHE-TAIL PCRpt_PT
dc.subjectnpp1pt_PT
dc.subjectPhytophthora cinnamomipt_PT
dc.subjectPichia pastorispt_PT
dc.subjectRT-qPCRpt_PT
dc.titleCloning, characterization, in vitro and in planta expression of a necrosis-inducing Phytophthora protein 1 gene npp1 from Phytophthora cinnamomipt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/3599-PPCDT/PTDC%2FAGR-AAM%2F67628%2F2006/PT
oaire.citation.endPage6462pt_PT
oaire.citation.issue6pt_PT
oaire.citation.startPage6453pt_PT
oaire.citation.titleMolecular Biology Reportspt_PT
oaire.citation.volume46pt_PT
oaire.fundingStream3599-PPCDT
person.familyNameCravador
person.givenNameAlfredo
person.identifier.orcid0000-0002-9831-9815
person.identifier.ridK-7247-2012
person.identifier.scopus-author-id6602537257
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsrestrictedAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublication8c84a9e6-6b7f-4b50-93a2-152c85901722
relation.isAuthorOfPublication.latestForDiscovery8c84a9e6-6b7f-4b50-93a2-152c85901722
relation.isProjectOfPublication9b7d24b8-1f01-4de4-ad2e-590347acee67
relation.isProjectOfPublication.latestForDiscovery9b7d24b8-1f01-4de4-ad2e-590347acee67

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