Cloning and expression analysis of an endo-1,3-β-D-glucosidase from Phytophthora cinnamomi.
Identifieur interne : 000272 ( Main/Exploration ); précédent : 000271; suivant : 000273Cloning and expression analysis of an endo-1,3-β-D-glucosidase from Phytophthora cinnamomi.
Auteurs : Rodrigo Costa [Portugal, Espagne] ; Angel Domínguez [Espagne] ; Altino Choupina [Portugal]Source :
- Molecular biology reports [ 1573-4978 ] ; 2020.
Descripteurs français
- KwdFr :
- Clonage moléculaire (méthodes), Glucan 1,3-beta-glucosidase (génétique), Glucan endo-1,3-beta-glucosidase (génétique), Glucan endo-1,3-beta-glucosidase (métabolisme), Glucosidases (génétique), Glucosidases (métabolisme), Maladies des plantes (MeSH), Phytophthora (génétique), Phytophthora (métabolisme), Réaction de polymérisation en chaine en temps réel (méthodes).
- MESH :
English descriptors
- KwdEn :
- Cloning, Molecular (methods), Glucan 1,3-beta-Glucosidase (genetics), Glucan Endo-1,3-beta-D-Glucosidase (genetics), Glucan Endo-1,3-beta-D-Glucosidase (metabolism), Glucosidases (genetics), Glucosidases (metabolism), Phytophthora (genetics), Phytophthora (metabolism), Plant Diseases (MeSH), Real-Time Polymerase Chain Reaction (methods).
- MESH :
- chemical , genetics : Glucan 1,3-beta-Glucosidase, Glucan Endo-1,3-beta-D-Glucosidase, Glucosidases.
- chemical , metabolism : Glucan Endo-1,3-beta-D-Glucosidase, Glucosidases.
- genetics : Phytophthora.
- metabolism : Phytophthora.
- methods : Cloning, Molecular, Real-Time Polymerase Chain Reaction.
- Plant Diseases.
Abstract
Phytophthora is considered one of the most destructive genus for many agricultural plant species worldwide, with a strong environmental and economic impact. Phytophthora cinnamomi is a highly aggressive Phytophthora species associated with the forest decline and responsible for the ink disease in chestnut trees (Castanea sativa Miller), a culture which is extremely important in Europe. This pathogenicity occurs due to the action of several enzymes like the hydrolysis of 1,3-β-glucans at specific sites by the enzyme endo-1,3-β-D-glucosidase. The aim of this work to analyze the heterologous expression in two microorganisms, Escherichia coli and Pichia pastoris, of an endo-1,3-β-D-glucosidase encoded by the gene ENDO1 (AM259651) from P. cinnamomi. Different plasmids were used to clone the gene on each organism and the real-time quantitative polymerase chain reaction was used to determine its level of expression. Homologous expression was also analyzed during growth in different carbon sources (glucose, cellulose, and sawdust) and time-course experiments were used for endo-1,3-β-D-glucosidase production. The highest expression of the endo-1,3-β-D-glucosidase gene occurred in glucose after 8 h of induction. In vivo infection of C. sativa by P. cinnamomi revealed an increase in endo-1,3-β-D-glucosidase expression after 12 h. At 24 h its expression decreased and at 48 h there was again a slight increase in expression, and more experiments in order to further explain this fact are underway.
DOI: 10.1007/s11033-019-05185-9
PubMed: 31741259
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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wicri:level="1"><nlm:affiliation>Centro de Investigação de Montanha (CIMO), Instituto Politécnico de Bragança, Campus de Santa Apolónia, 5300-253, Bragança, Portugal. albracho@ipb.pt.</nlm:affiliation><country xml:lang="fr">Portugal</country><wicri:regionArea>Centro de Investigação de Montanha (CIMO), Instituto Politécnico de Bragança, Campus de Santa Apolónia, 5300-253, Bragança</wicri:regionArea><wicri:noRegion>Bragança</wicri:noRegion></affiliation></author></analytic><series><title level="j">Molecular biology reports</title><idno type="eISSN">1573-4978</idno><imprint><date when="2020" type="published">2020</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Cloning, Molecular (methods)</term><term>Glucan 1,3-beta-Glucosidase (genetics)</term><term>Glucan Endo-1,3-beta-D-Glucosidase (genetics)</term><term>Glucan Endo-1,3-beta-D-Glucosidase (metabolism)</term><term>Glucosidases (genetics)</term><term>Glucosidases (metabolism)</term><term>Phytophthora (genetics)</term><term>Phytophthora (metabolism)</term><term>Plant Diseases (MeSH)</term><term>Real-Time Polymerase Chain Reaction (methods)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Clonage moléculaire (méthodes)</term><term>Glucan 1,3-beta-glucosidase (génétique)</term><term>Glucan endo-1,3-beta-glucosidase (génétique)</term><term>Glucan endo-1,3-beta-glucosidase (métabolisme)</term><term>Glucosidases (génétique)</term><term>Glucosidases (métabolisme)</term><term>Maladies des plantes (MeSH)</term><term>Phytophthora (génétique)</term><term>Phytophthora (métabolisme)</term><term>Réaction de polymérisation en chaine en temps réel (méthodes)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Glucan 1,3-beta-Glucosidase</term><term>Glucan Endo-1,3-beta-D-Glucosidase</term><term>Glucosidases</term></keywords><keywords scheme="MESH" type="chemical" 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réel</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Plant Diseases</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Maladies des plantes</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Phytophthora is considered one of the most destructive genus for many agricultural plant species worldwide, with a strong environmental and economic impact. Phytophthora cinnamomi is a highly aggressive Phytophthora species associated with the forest decline and responsible for the ink disease in chestnut trees (Castanea sativa Miller), a culture which is extremely important in Europe. This pathogenicity occurs due to the action of several enzymes like the hydrolysis of 1,3-β-glucans at specific sites by the enzyme endo-1,3-β-D-glucosidase. The aim of this work to analyze the heterologous expression in two microorganisms, Escherichia coli and Pichia pastoris, of an endo-1,3-β-D-glucosidase encoded by the gene ENDO1 (AM259651) from P. cinnamomi. Different plasmids were used to clone the gene on each organism and the real-time quantitative polymerase chain reaction was used to determine its level of expression. Homologous expression was also analyzed during growth in different carbon sources (glucose, cellulose, and sawdust) and time-course experiments were used for endo-1,3-β-D-glucosidase production. The highest expression of the endo-1,3-β-D-glucosidase gene occurred in glucose after 8 h of induction. In vivo infection of C. sativa by P. cinnamomi revealed an increase in endo-1,3-β-D-glucosidase expression after 12 h. At 24 h its expression decreased and at 48 h there was again a slight increase in expression, and more experiments in order to further explain this fact are underway.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">31741259</PMID><DateCompleted><Year>2020</Year><Month>06</Month><Day>23</Day></DateCompleted><DateRevised><Year>2020</Year><Month>10</Month><Day>27</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1573-4978</ISSN><JournalIssue CitedMedium="Internet"><Volume>47</Volume><Issue>2</Issue><PubDate><Year>2020</Year><Month>Feb</Month></PubDate></JournalIssue><Title>Molecular biology reports</Title><ISOAbbreviation>Mol Biol Rep</ISOAbbreviation></Journal><ArticleTitle>Cloning and expression analysis of an endo-1,3-β-D-glucosidase from Phytophthora cinnamomi.</ArticleTitle><Pagination><MedlinePgn>935-942</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1007/s11033-019-05185-9</ELocationID><Abstract><AbstractText>Phytophthora is considered one of the most destructive genus for many agricultural plant species worldwide, with a strong environmental and economic impact. Phytophthora cinnamomi is a highly aggressive Phytophthora species associated with the forest decline and responsible for the ink disease in chestnut trees (Castanea sativa Miller), a culture which is extremely important in Europe. This pathogenicity occurs due to the action of several enzymes like the hydrolysis of 1,3-β-glucans at specific sites by the enzyme endo-1,3-β-D-glucosidase. The aim of this work to analyze the heterologous expression in two microorganisms, Escherichia coli and Pichia pastoris, of an endo-1,3-β-D-glucosidase encoded by the gene ENDO1 (AM259651) from P. cinnamomi. Different plasmids were used to clone the gene on each organism and the real-time quantitative polymerase chain reaction was used to determine its level of expression. Homologous expression was also analyzed during growth in different carbon sources (glucose, cellulose, and sawdust) and time-course experiments were used for endo-1,3-β-D-glucosidase production. The highest expression of the endo-1,3-β-D-glucosidase gene occurred in glucose after 8 h of induction. In vivo infection of C. sativa by P. cinnamomi revealed an increase in endo-1,3-β-D-glucosidase expression after 12 h. At 24 h its expression decreased and at 48 h there was again a slight increase in expression, and more experiments in order to further explain this fact are underway.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Costa</LastName><ForeName>Rodrigo</ForeName><Initials>R</Initials><AffiliationInfo><Affiliation>Centro de Investigação de Montanha (CIMO), Instituto Politécnico de Bragança, Campus de Santa Apolónia, 5300-253, Bragança, Portugal.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Departamento de Microbiología y Genética, Edificio Departamental, Plaza de los Drs. de la Reina s/n, 37007, Salamanca, Spain.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Domínguez</LastName><ForeName>Angel</ForeName><Initials>A</Initials><AffiliationInfo><Affiliation>Departamento de Microbiología y Genética, Edificio Departamental, Plaza de los Drs. de la Reina s/n, 37007, Salamanca, Spain.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Choupina</LastName><ForeName>Altino</ForeName><Initials>A</Initials><Identifier Source="ORCID">http://orcid.org/0000-0002-3956-9398</Identifier><AffiliationInfo><Affiliation>Centro de Investigação de Montanha (CIMO), Instituto Politécnico de Bragança, Campus de Santa Apolónia, 5300-253, Bragança, Portugal. albracho@ipb.pt.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2019</Year><Month>11</Month><Day>18</Day></ArticleDate></Article><MedlineJournalInfo><Country>Netherlands</Country><MedlineTA>Mol Biol Rep</MedlineTA><NlmUniqueID>0403234</NlmUniqueID><ISSNLinking>0301-4851</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>EC 3.2.1.-</RegistryNumber><NameOfSubstance UI="D005959">Glucosidases</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 3.2.1.39</RegistryNumber><NameOfSubstance UI="D004693">Glucan Endo-1,3-beta-D-Glucosidase</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 3.2.1.58</RegistryNumber><NameOfSubstance UI="D043326">Glucan 1,3-beta-Glucosidase</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D003001" MajorTopicYN="N">Cloning, Molecular</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="N">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D043326" MajorTopicYN="N">Glucan 1,3-beta-Glucosidase</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D004693" MajorTopicYN="N">Glucan Endo-1,3-beta-D-Glucosidase</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D005959" MajorTopicYN="N">Glucosidases</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010838" MajorTopicYN="N">Phytophthora</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010935" MajorTopicYN="N">Plant Diseases</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D060888" MajorTopicYN="N">Real-Time Polymerase Chain Reaction</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="N">methods</QualifierName></MeshHeading></MeshHeadingList><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">Castanea sativa</Keyword><Keyword MajorTopicYN="N">Endo-1,3-β-D-glucosidase</Keyword><Keyword MajorTopicYN="N">Heterologous expression</Keyword><Keyword MajorTopicYN="N">Homologous expression</Keyword><Keyword MajorTopicYN="N">Phytophthora cinnamomi</Keyword><Keyword MajorTopicYN="N">RT-qPCR</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2019</Year><Month>03</Month><Day>26</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2019</Year><Month>11</Month><Day>07</Day></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2019</Year><Month>11</Month><Day>20</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2020</Year><Month>6</Month><Day>24</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2019</Year><Month>11</Month><Day>20</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">31741259</ArticleId><ArticleId IdType="doi">10.1007/s11033-019-05185-9</ArticleId><ArticleId IdType="pii">10.1007/s11033-019-05185-9</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Espagne</li><li>Portugal</li></country></list><tree><country name="Portugal"><noRegion><name sortKey="Costa, Rodrigo" sort="Costa, Rodrigo" uniqKey="Costa R" first="Rodrigo" last="Costa">Rodrigo Costa</name></noRegion><name sortKey="Choupina, Altino" sort="Choupina, Altino" uniqKey="Choupina A" first="Altino" last="Choupina">Altino Choupina</name></country><country name="Espagne"><noRegion><name sortKey="Costa, Rodrigo" sort="Costa, Rodrigo" uniqKey="Costa R" first="Rodrigo" last="Costa">Rodrigo Costa</name></noRegion><name sortKey="Dominguez, Angel" sort="Dominguez, Angel" uniqKey="Dominguez A" first="Angel" last="Domínguez">Angel Domínguez</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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