RustEffectorV1, Main, Exploration, bibRecord, 000110

A bacterial type III secretion assay for delivery of fungal effector proteins into wheat.

Identifieur interne : 000110 ( Main/Exploration ); précédent : 000109; suivant : 000111

A bacterial type III secretion assay for delivery of fungal effector proteins into wheat.

Auteurs : Narayana M. Upadhyaya ; Rohit Mago ; Brian J. Staskawicz ; Michael A. Ayliffe ; Jeffrey G. Ellis ; Peter N. Dodds

Source :

Molecular plant-microbe interactions : MPMI [ 0894-0282 ] ; 2014.

RBID : pubmed:24156769

Descripteurs français

English descriptors

KwdEn :
- Adenylyl Cyclases (genetics), Adenylyl Cyclases (metabolism), Bacterial Proteins (genetics), Bacterial Proteins (metabolism), Basidiomycota (pathogenicity), Calmodulin (genetics), Calmodulin (metabolism), Genetic Engineering (MeSH), Hordeum (metabolism), Hordeum (microbiology), Plant Diseases (microbiology), Plant Leaves (metabolism), Plant Leaves (microbiology), Plant Stems (metabolism), Plant Stems (microbiology), Plants, Genetically Modified (MeSH), Protein Transport (MeSH), Pseudomonas fluorescens (genetics), Pseudomonas fluorescens (metabolism), Pseudomonas syringae (genetics), Recombinant Fusion Proteins (MeSH), Triticum (metabolism), Triticum (microbiology), Virulence (MeSH), Xanthomonas (genetics).
MESH :
- chemical , genetics : Adenylyl Cyclases, Bacterial Proteins, Calmodulin.
- chemical , metabolism : Adenylyl Cyclases, Bacterial Proteins, Calmodulin.
- genetics : Pseudomonas fluorescens, Pseudomonas syringae, Xanthomonas.
- metabolism : Hordeum, Plant Leaves, Plant Stems, Pseudomonas fluorescens, Triticum.
- microbiology : Hordeum, Plant Diseases, Plant Leaves, Plant Stems, Triticum.
- pathogenicity : Basidiomycota.
- Genetic Engineering, Plants, Genetically Modified, Protein Transport, Recombinant Fusion Proteins, Virulence.

Abstract

Large numbers of candidate effectors from fungal pathogens are being identified through whole-genome sequencing and in planta expression studies. Although Agrobacterium-mediated transient expression has enabled high-throughput functional analysis of effectors in dicot plants, this assay is not effective in cereal leaves. Here, we show that a nonpathogenic Pseudomonas fluorescens engineered to express the type III secretion system (T3SS) of P. syringae and the wheat pathogen Xanthomonas translucens can deliver fusion proteins containing T3SS signals from P. syringae (AvrRpm1) and X. campestris (AvrBs2) avirulence (Avr) proteins, respectively, into wheat leaf cells. A calmodulin-dependent adenylate cyclase reporter protein was delivered effectively into wheat and barley by both bacteria. Absence of any disease symptoms with P. fluorescens makes it more suitable than X. translucens for detecting a hypersensitive response (HR) induced by an effector protein with avirulence activity. We further modified the delivery system by removal of the myristoylation site from the AvrRpm1 fusion to prevent its localization to the plasma membrane which could inhibit recognition of an Avr protein. Delivery of the flax rust AvrM protein by the modified delivery system into transgenic tobacco leaves expressing the corresponding M resistance protein induced a strong HR, indicating that the system is capable of delivering a functional rust Avr protein. In a preliminary screen of effectors from the stem rust fungus Puccinia graminis f. sp. tritici, we identified one effector that induced a host genotype-specific HR in wheat. Thus, the modified AvrRpm1:effector-Pseudomonas fluorescens system is an effective tool for large-scale screening of pathogen effectors for recognition in wheat.

DOI: 10.1094/MPMI-07-13-0187-FI
PubMed: 24156769

Affiliations:

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Le document en format XML

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<author><name sortKey="Upadhyaya, Narayana M" sort="Upadhyaya, Narayana M" uniqKey="Upadhyaya N" first="Narayana M" last="Upadhyaya">Narayana M. Upadhyaya</name>
</author>
<author><name sortKey="Mago, Rohit" sort="Mago, Rohit" uniqKey="Mago R" first="Rohit" last="Mago">Rohit Mago</name>
</author>
<author><name sortKey="Staskawicz, Brian J" sort="Staskawicz, Brian J" uniqKey="Staskawicz B" first="Brian J" last="Staskawicz">Brian J. Staskawicz</name>
</author>
<author><name sortKey="Ayliffe, Michael A" sort="Ayliffe, Michael A" uniqKey="Ayliffe M" first="Michael A" last="Ayliffe">Michael A. Ayliffe</name>
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<author><name sortKey="Ellis, Jeffrey G" sort="Ellis, Jeffrey G" uniqKey="Ellis J" first="Jeffrey G" last="Ellis">Jeffrey G. Ellis</name>
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<author><name sortKey="Dodds, Peter N" sort="Dodds, Peter N" uniqKey="Dodds P" first="Peter N" last="Dodds">Peter N. Dodds</name>
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<sourceDesc><biblStruct><analytic><title xml:lang="en">A bacterial type III secretion assay for delivery of fungal effector proteins into wheat.</title>
<author><name sortKey="Upadhyaya, Narayana M" sort="Upadhyaya, Narayana M" uniqKey="Upadhyaya N" first="Narayana M" last="Upadhyaya">Narayana M. Upadhyaya</name>
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<author><name sortKey="Mago, Rohit" sort="Mago, Rohit" uniqKey="Mago R" first="Rohit" last="Mago">Rohit Mago</name>
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<author><name sortKey="Staskawicz, Brian J" sort="Staskawicz, Brian J" uniqKey="Staskawicz B" first="Brian J" last="Staskawicz">Brian J. Staskawicz</name>
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<author><name sortKey="Ayliffe, Michael A" sort="Ayliffe, Michael A" uniqKey="Ayliffe M" first="Michael A" last="Ayliffe">Michael A. Ayliffe</name>
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<author><name sortKey="Ellis, Jeffrey G" sort="Ellis, Jeffrey G" uniqKey="Ellis J" first="Jeffrey G" last="Ellis">Jeffrey G. Ellis</name>
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<author><name sortKey="Dodds, Peter N" sort="Dodds, Peter N" uniqKey="Dodds P" first="Peter N" last="Dodds">Peter N. Dodds</name>
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<series><title level="j">Molecular plant-microbe interactions : MPMI</title>
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<imprint><date when="2014" type="published">2014</date>
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Adenylyl Cyclases (genetics)</term>
<term>Adenylyl Cyclases (metabolism)</term>
<term>Bacterial Proteins (genetics)</term>
<term>Bacterial Proteins (metabolism)</term>
<term>Basidiomycota (pathogenicity)</term>
<term>Calmodulin (genetics)</term>
<term>Calmodulin (metabolism)</term>
<term>Genetic Engineering (MeSH)</term>
<term>Hordeum (metabolism)</term>
<term>Hordeum (microbiology)</term>
<term>Plant Diseases (microbiology)</term>
<term>Plant Leaves (metabolism)</term>
<term>Plant Leaves (microbiology)</term>
<term>Plant Stems (metabolism)</term>
<term>Plant Stems (microbiology)</term>
<term>Plants, Genetically Modified (MeSH)</term>
<term>Protein Transport (MeSH)</term>
<term>Pseudomonas fluorescens (genetics)</term>
<term>Pseudomonas fluorescens (metabolism)</term>
<term>Pseudomonas syringae (genetics)</term>
<term>Recombinant Fusion Proteins (MeSH)</term>
<term>Triticum (metabolism)</term>
<term>Triticum (microbiology)</term>
<term>Virulence (MeSH)</term>
<term>Xanthomonas (genetics)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Adenylate Cyclase (génétique)</term>
<term>Adenylate Cyclase (métabolisme)</term>
<term>Basidiomycota (pathogénicité)</term>
<term>Calmoduline (génétique)</term>
<term>Calmoduline (métabolisme)</term>
<term>Feuilles de plante (microbiologie)</term>
<term>Feuilles de plante (métabolisme)</term>
<term>Génie génétique (MeSH)</term>
<term>Hordeum (microbiologie)</term>
<term>Hordeum (métabolisme)</term>
<term>Maladies des plantes (microbiologie)</term>
<term>Protéines bactériennes (génétique)</term>
<term>Protéines bactériennes (métabolisme)</term>
<term>Protéines de fusion recombinantes (MeSH)</term>
<term>Pseudomonas fluorescens (génétique)</term>
<term>Pseudomonas fluorescens (métabolisme)</term>
<term>Pseudomonas syringae (génétique)</term>
<term>Tiges de plante (microbiologie)</term>
<term>Tiges de plante (métabolisme)</term>
<term>Transport des protéines (MeSH)</term>
<term>Triticum (microbiologie)</term>
<term>Triticum (métabolisme)</term>
<term>Virulence (MeSH)</term>
<term>Végétaux génétiquement modifiés (MeSH)</term>
<term>Xanthomonas (génétique)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Adenylyl Cyclases</term>
<term>Bacterial Proteins</term>
<term>Calmodulin</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Adenylyl Cyclases</term>
<term>Bacterial Proteins</term>
<term>Calmodulin</term>
</keywords>
<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Pseudomonas fluorescens</term>
<term>Pseudomonas syringae</term>
<term>Xanthomonas</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Adenylate Cyclase</term>
<term>Calmoduline</term>
<term>Protéines bactériennes</term>
<term>Pseudomonas fluorescens</term>
<term>Pseudomonas syringae</term>
<term>Xanthomonas</term>
</keywords>
<keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Hordeum</term>
<term>Plant Leaves</term>
<term>Plant Stems</term>
<term>Pseudomonas fluorescens</term>
<term>Triticum</term>
</keywords>
<keywords scheme="MESH" qualifier="microbiologie" xml:lang="fr"><term>Feuilles de plante</term>
<term>Hordeum</term>
<term>Maladies des plantes</term>
<term>Tiges de plante</term>
<term>Triticum</term>
</keywords>
<keywords scheme="MESH" qualifier="microbiology" xml:lang="en"><term>Hordeum</term>
<term>Plant Diseases</term>
<term>Plant Leaves</term>
<term>Plant Stems</term>
<term>Triticum</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Adenylate Cyclase</term>
<term>Calmoduline</term>
<term>Feuilles de plante</term>
<term>Hordeum</term>
<term>Protéines bactériennes</term>
<term>Pseudomonas fluorescens</term>
<term>Tiges de plante</term>
<term>Triticum</term>
</keywords>
<keywords scheme="MESH" qualifier="pathogenicity" xml:lang="en"><term>Basidiomycota</term>
</keywords>
<keywords scheme="MESH" qualifier="pathogénicité" xml:lang="fr"><term>Basidiomycota</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Genetic Engineering</term>
<term>Plants, Genetically Modified</term>
<term>Protein Transport</term>
<term>Recombinant Fusion Proteins</term>
<term>Virulence</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Génie génétique</term>
<term>Protéines de fusion recombinantes</term>
<term>Transport des protéines</term>
<term>Virulence</term>
<term>Végétaux génétiquement modifiés</term>
</keywords>
</textClass>
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</teiHeader>
<front><div type="abstract" xml:lang="en">Large numbers of candidate effectors from fungal pathogens are being identified through whole-genome sequencing and in planta expression studies. Although Agrobacterium-mediated transient expression has enabled high-throughput functional analysis of effectors in dicot plants, this assay is not effective in cereal leaves. Here, we show that a nonpathogenic Pseudomonas fluorescens engineered to express the type III secretion system (T3SS) of P. syringae and the wheat pathogen Xanthomonas translucens can deliver fusion proteins containing T3SS signals from P. syringae (AvrRpm1) and X. campestris (AvrBs2) avirulence (Avr) proteins, respectively, into wheat leaf cells. A calmodulin-dependent adenylate cyclase reporter protein was delivered effectively into wheat and barley by both bacteria. Absence of any disease symptoms with P. fluorescens makes it more suitable than X. translucens for detecting a hypersensitive response (HR) induced by an effector protein with avirulence activity. We further modified the delivery system by removal of the myristoylation site from the AvrRpm1 fusion to prevent its localization to the plasma membrane which could inhibit recognition of an Avr protein. Delivery of the flax rust AvrM protein by the modified delivery system into transgenic tobacco leaves expressing the corresponding M resistance protein induced a strong HR, indicating that the system is capable of delivering a functional rust Avr protein. In a preliminary screen of effectors from the stem rust fungus Puccinia graminis f. sp. tritici, we identified one effector that induced a host genotype-specific HR in wheat. Thus, the modified AvrRpm1:effector-Pseudomonas fluorescens system is an effective tool for large-scale screening of pathogen effectors for recognition in wheat. </div>
</front>
</TEI>
<pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">24156769</PMID>
<DateCompleted><Year>2014</Year>
<Month>04</Month>
<Day>24</Day>
</DateCompleted>
<DateRevised><Year>2015</Year>
<Month>11</Month>
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<Issue>3</Issue>
<PubDate><Year>2014</Year>
<Month>Mar</Month>
</PubDate>
</JournalIssue>
<Title>Molecular plant-microbe interactions : MPMI</Title>
<ISOAbbreviation>Mol Plant Microbe Interact</ISOAbbreviation>
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<ArticleTitle>A bacterial type III secretion assay for delivery of fungal effector proteins into wheat.</ArticleTitle>
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<ELocationID EIdType="doi" ValidYN="Y">10.1094/MPMI-07-13-0187-FI</ELocationID>
<Abstract><AbstractText>Large numbers of candidate effectors from fungal pathogens are being identified through whole-genome sequencing and in planta expression studies. Although Agrobacterium-mediated transient expression has enabled high-throughput functional analysis of effectors in dicot plants, this assay is not effective in cereal leaves. Here, we show that a nonpathogenic Pseudomonas fluorescens engineered to express the type III secretion system (T3SS) of P. syringae and the wheat pathogen Xanthomonas translucens can deliver fusion proteins containing T3SS signals from P. syringae (AvrRpm1) and X. campestris (AvrBs2) avirulence (Avr) proteins, respectively, into wheat leaf cells. A calmodulin-dependent adenylate cyclase reporter protein was delivered effectively into wheat and barley by both bacteria. Absence of any disease symptoms with P. fluorescens makes it more suitable than X. translucens for detecting a hypersensitive response (HR) induced by an effector protein with avirulence activity. We further modified the delivery system by removal of the myristoylation site from the AvrRpm1 fusion to prevent its localization to the plasma membrane which could inhibit recognition of an Avr protein. Delivery of the flax rust AvrM protein by the modified delivery system into transgenic tobacco leaves expressing the corresponding M resistance protein induced a strong HR, indicating that the system is capable of delivering a functional rust Avr protein. In a preliminary screen of effectors from the stem rust fungus Puccinia graminis f. sp. tritici, we identified one effector that induced a host genotype-specific HR in wheat. Thus, the modified AvrRpm1:effector-Pseudomonas fluorescens system is an effective tool for large-scale screening of pathogen effectors for recognition in wheat. </AbstractText>
</Abstract>
<AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Upadhyaya</LastName>
<ForeName>Narayana M</ForeName>
<Initials>NM</Initials>
</Author>
<Author ValidYN="Y"><LastName>Mago</LastName>
<ForeName>Rohit</ForeName>
<Initials>R</Initials>
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<Author ValidYN="Y"><LastName>Staskawicz</LastName>
<ForeName>Brian J</ForeName>
<Initials>BJ</Initials>
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<Author ValidYN="Y"><LastName>Ayliffe</LastName>
<ForeName>Michael A</ForeName>
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</Author>
<Author ValidYN="Y"><LastName>Ellis</LastName>
<ForeName>Jeffrey G</ForeName>
<Initials>JG</Initials>
</Author>
<Author ValidYN="Y"><LastName>Dodds</LastName>
<ForeName>Peter N</ForeName>
<Initials>PN</Initials>
</Author>
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<Language>eng</Language>
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<ChemicalList><Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="C417273">AvrBs2 protein, Xanthomonas campestris</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="C502082">AvrRpm1 protein, Pseudomonas syringae</NameOfSubstance>
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<Chemical><RegistryNumber>EC 4.6.1.1</RegistryNumber>
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<CitationSubset>IM</CitationSubset>
<MeshHeadingList><MeshHeading><DescriptorName UI="D000262" MajorTopicYN="N">Adenylyl Cyclases</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName>
<QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName>
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<MeshHeading><DescriptorName UI="D001426" MajorTopicYN="N">Bacterial Proteins</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName>
<QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName>
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<MeshHeading><DescriptorName UI="D001487" MajorTopicYN="N">Basidiomycota</DescriptorName>
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<QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName>
<QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName>
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<MeshHeading><DescriptorName UI="D005818" MajorTopicYN="N">Genetic Engineering</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D001467" MajorTopicYN="N">Hordeum</DescriptorName>
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<QualifierName UI="Q000382" MajorTopicYN="N">microbiology</QualifierName>
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<MeshHeading><DescriptorName UI="D010935" MajorTopicYN="N">Plant Diseases</DescriptorName>
<QualifierName UI="Q000382" MajorTopicYN="Y">microbiology</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D018515" MajorTopicYN="N">Plant Leaves</DescriptorName>
<QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName>
<QualifierName UI="Q000382" MajorTopicYN="N">microbiology</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D018547" MajorTopicYN="N">Plant Stems</DescriptorName>
<QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName>
<QualifierName UI="Q000382" MajorTopicYN="N">microbiology</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D030821" MajorTopicYN="N">Plants, Genetically Modified</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D021381" MajorTopicYN="N">Protein Transport</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D011551" MajorTopicYN="N">Pseudomonas fluorescens</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName>
<QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D044224" MajorTopicYN="N">Pseudomonas syringae</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D011993" MajorTopicYN="N">Recombinant Fusion Proteins</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D014908" MajorTopicYN="N">Triticum</DescriptorName>
<QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName>
<QualifierName UI="Q000382" MajorTopicYN="N">microbiology</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D014774" MajorTopicYN="N">Virulence</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D014974" MajorTopicYN="N">Xanthomonas</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName>
</MeshHeading>
</MeshHeadingList>
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<PubmedData><History><PubMedPubDate PubStatus="entrez"><Year>2013</Year>
<Month>10</Month>
<Day>26</Day>
<Hour>6</Hour>
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<tree><noCountry><name sortKey="Ayliffe, Michael A" sort="Ayliffe, Michael A" uniqKey="Ayliffe M" first="Michael A" last="Ayliffe">Michael A. Ayliffe</name>
<name sortKey="Dodds, Peter N" sort="Dodds, Peter N" uniqKey="Dodds P" first="Peter N" last="Dodds">Peter N. Dodds</name>
<name sortKey="Ellis, Jeffrey G" sort="Ellis, Jeffrey G" uniqKey="Ellis J" first="Jeffrey G" last="Ellis">Jeffrey G. Ellis</name>
<name sortKey="Mago, Rohit" sort="Mago, Rohit" uniqKey="Mago R" first="Rohit" last="Mago">Rohit Mago</name>
<name sortKey="Staskawicz, Brian J" sort="Staskawicz, Brian J" uniqKey="Staskawicz B" first="Brian J" last="Staskawicz">Brian J. Staskawicz</name>
<name sortKey="Upadhyaya, Narayana M" sort="Upadhyaya, Narayana M" uniqKey="Upadhyaya N" first="Narayana M" last="Upadhyaya">Narayana M. Upadhyaya</name>
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A bacterial type III secretion assay for delivery of fungal effector proteins into wheat.

A bacterial type III secretion assay for delivery of fungal effector proteins into wheat.

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Descripteurs français

English descriptors

Abstract

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