Agromonas: a rapid disease assay for Pseudomonas syringae growth in agroinfiltrated leaves.
Identifieur interne : 000220 ( Main/Exploration ); précédent : 000219; suivant : 000221Agromonas: a rapid disease assay for Pseudomonas syringae growth in agroinfiltrated leaves.
Auteurs : Pierre Buscaill [Royaume-Uni] ; Nattapong Sanguankiattichai [Royaume-Uni] ; Yoon Joo Lee [Royaume-Uni] ; Jiorgos Kourelis [Royaume-Uni] ; Gail Preston [Royaume-Uni] ; Renier A L. Van Der Hoorn [Royaume-Uni]Source :
- The Plant journal : for cell and molecular biology [ 1365-313X ] ; 2020.
Abstract
The lengthy process to generate transformed plants is a limitation in current research on the interactions of the model plant pathogen Pseudomonas syringae with plant hosts. Here we present an easy method called agromonas, where we quantify P. syringae growth in agroinfiltrated leaves of Nicotiana benthamiana using a cocktail of antibiotics to select P. syringae on plates. As a proof of concept, we demonstrate that transient expression of PAMP receptors reduces bacterial growth and that transient depletion of a host immune gene and transient expression of a type-III effector increase P. syringae growth in agromonas assays. We show that we can rapidly achieve structure-function analysis of immune components and test the function of immune hydrolases. The agromonas method is easy, fast and robust for routine disease assays with various Pseudomonas strains without transforming plants or bacteria. The agromonas assay offers a reliable approach for further comprehensive analysis of plant immunity. Supporting Information Table S1 Selection of bacterial strains on medium supplemented with CFC or gentamicin. Table S2 Binary vectors generated in this study. Table S3 Primers and nucleic acid sequences used in this study. Table S4 Bacterial strains used in this study.
DOI: 10.1111/tpj.15056
PubMed: 33124734
Affiliations:
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Van Der Hoorn</name><affiliation wicri:level="4"><nlm:affiliation>Plant Chemetics lab, Department of Plant Sciences, University of Oxford, OX1 3RB, Oxford, UK.</nlm:affiliation><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Plant Chemetics lab, Department of Plant Sciences, University of Oxford, OX1 3RB, Oxford</wicri:regionArea><placeName><settlement type="city">Oxford</settlement><region type="country">Angleterre</region><region type="comté" nuts="2">Oxfordshire</region></placeName><orgName type="university">Université d'Oxford</orgName></affiliation></author></analytic><series><title level="j">The Plant journal : for cell and molecular biology</title><idno type="eISSN">1365-313X</idno><imprint><date when="2020" type="published">2020</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The lengthy process to generate transformed plants is a limitation in current research on the interactions of the model plant pathogen Pseudomonas syringae with plant hosts. Here we present an easy method called agromonas, where we quantify P. syringae growth in agroinfiltrated leaves of Nicotiana benthamiana using a cocktail of antibiotics to select P. syringae on plates. As a proof of concept, we demonstrate that transient expression of PAMP receptors reduces bacterial growth and that transient depletion of a host immune gene and transient expression of a type-III effector increase P. syringae growth in agromonas assays. We show that we can rapidly achieve structure-function analysis of immune components and test the function of immune hydrolases. The agromonas method is easy, fast and robust for routine disease assays with various Pseudomonas strains without transforming plants or bacteria. The agromonas assay offers a reliable approach for further comprehensive analysis of plant immunity. Supporting Information Table S1 Selection of bacterial strains on medium supplemented with CFC or gentamicin. Table S2 Binary vectors generated in this study. Table S3 Primers and nucleic acid sequences used in this study. Table S4 Bacterial strains used in this study.</div></front></TEI><pubmed><MedlineCitation Status="Publisher" Owner="NLM"><PMID Version="1">33124734</PMID><DateRevised><Year>2020</Year><Month>10</Month><Day>30</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1365-313X</ISSN><JournalIssue CitedMedium="Internet"><PubDate><Year>2020</Year><Month>Oct</Month><Day>30</Day></PubDate></JournalIssue><Title>The Plant journal : for cell and molecular biology</Title><ISOAbbreviation>Plant J</ISOAbbreviation></Journal><ArticleTitle>Agromonas: a rapid disease assay for Pseudomonas syringae growth in agroinfiltrated leaves.</ArticleTitle><ELocationID EIdType="doi" ValidYN="Y">10.1111/tpj.15056</ELocationID><Abstract><AbstractText>The lengthy process to generate transformed plants is a limitation in current research on the interactions of the model plant pathogen Pseudomonas syringae with plant hosts. Here we present an easy method called agromonas, where we quantify P. syringae growth in agroinfiltrated leaves of Nicotiana benthamiana using a cocktail of antibiotics to select P. syringae on plates. As a proof of concept, we demonstrate that transient expression of PAMP receptors reduces bacterial growth and that transient depletion of a host immune gene and transient expression of a type-III effector increase P. syringae growth in agromonas assays. We show that we can rapidly achieve structure-function analysis of immune components and test the function of immune hydrolases. The agromonas method is easy, fast and robust for routine disease assays with various Pseudomonas strains without transforming plants or bacteria. The agromonas assay offers a reliable approach for further comprehensive analysis of plant immunity. Supporting Information Table S1 Selection of bacterial strains on medium supplemented with CFC or gentamicin. Table S2 Binary vectors generated in this study. Table S3 Primers and nucleic acid sequences used in this study. Table S4 Bacterial strains used in this study.</AbstractText><CopyrightInformation>This article is protected by copyright. All rights reserved.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Buscaill</LastName><ForeName>Pierre</ForeName><Initials>P</Initials><AffiliationInfo><Affiliation>Plant Chemetics lab, Department of Plant Sciences, University of Oxford, OX1 3RB, Oxford, UK.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Sanguankiattichai</LastName><ForeName>Nattapong</ForeName><Initials>N</Initials><AffiliationInfo><Affiliation>Plant Chemetics lab, Department of Plant Sciences, University of Oxford, OX1 3RB, Oxford, UK.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Department of Plant Sciences, University of Oxford, OX1 3RB, Oxford, UK.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Joo Lee</LastName><ForeName>Yoon</ForeName><Initials>Y</Initials><AffiliationInfo><Affiliation>Plant Chemetics lab, Department of Plant Sciences, University of Oxford, OX1 3RB, Oxford, UK.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Kourelis</LastName><ForeName>Jiorgos</ForeName><Initials>J</Initials><Identifier Source="ORCID">https://orcid.org/0000-0002-9007-1333</Identifier><AffiliationInfo><Affiliation>Plant Chemetics lab, Department of Plant Sciences, University of Oxford, OX1 3RB, Oxford, UK.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Preston</LastName><ForeName>Gail</ForeName><Initials>G</Initials><AffiliationInfo><Affiliation>Department of Plant Sciences, University of Oxford, OX1 3RB, Oxford, UK.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>van der Hoorn</LastName><ForeName>Renier A L</ForeName><Initials>RAL</Initials><Identifier Source="ORCID">https://orcid.org/0000-0002-3692-7487</Identifier><AffiliationInfo><Affiliation>Plant Chemetics lab, Department of Plant Sciences, University of Oxford, OX1 3RB, Oxford, UK.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2020</Year><Month>10</Month><Day>30</Day></ArticleDate></Article><MedlineJournalInfo><Country>England</Country><MedlineTA>Plant J</MedlineTA><NlmUniqueID>9207397</NlmUniqueID><ISSNLinking>0960-7412</ISSNLinking></MedlineJournalInfo><CitationSubset>IM</CitationSubset><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">Agrobacterium</Keyword><Keyword MajorTopicYN="N">Nicotiana benthamiana</Keyword><Keyword MajorTopicYN="N">Pseudomonas syringae</Keyword><Keyword MajorTopicYN="N">disease assay</Keyword><Keyword MajorTopicYN="N">plant immunity</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="entrez"><Year>2020</Year><Month>10</Month><Day>30</Day><Hour>8</Hour><Minute>39</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2020</Year><Month>10</Month><Day>31</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2020</Year><Month>10</Month><Day>31</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>aheadofprint</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">33124734</ArticleId><ArticleId IdType="doi">10.1111/tpj.15056</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Royaume-Uni</li></country><region><li>Angleterre</li><li>Oxfordshire</li></region><settlement><li>Oxford</li></settlement><orgName><li>Université d'Oxford</li></orgName></list><tree><country name="Royaume-Uni"><region name="Angleterre"><name sortKey="Buscaill, Pierre" sort="Buscaill, Pierre" uniqKey="Buscaill P" first="Pierre" last="Buscaill">Pierre Buscaill</name></region><name sortKey="Joo Lee, Yoon" sort="Joo Lee, Yoon" uniqKey="Joo Lee Y" first="Yoon" last="Joo Lee">Yoon Joo Lee</name><name sortKey="Kourelis, Jiorgos" sort="Kourelis, Jiorgos" uniqKey="Kourelis J" first="Jiorgos" last="Kourelis">Jiorgos Kourelis</name><name sortKey="Preston, Gail" sort="Preston, Gail" uniqKey="Preston G" first="Gail" last="Preston">Gail Preston</name><name sortKey="Sanguankiattichai, Nattapong" sort="Sanguankiattichai, Nattapong" uniqKey="Sanguankiattichai N" first="Nattapong" last="Sanguankiattichai">Nattapong Sanguankiattichai</name><name sortKey="Sanguankiattichai, Nattapong" sort="Sanguankiattichai, Nattapong" uniqKey="Sanguankiattichai N" first="Nattapong" last="Sanguankiattichai">Nattapong Sanguankiattichai</name><name sortKey="Van Der Hoorn, Renier A L" sort="Van Der Hoorn, Renier A L" uniqKey="Van Der Hoorn R" first="Renier A L" last="Van Der Hoorn">Renier A L. 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