Optimization of Agroinfiltration in Pisum sativum Provides a New Tool for Studying the Salivary Protein Functions in the Pea Aphid Complex.
Identifieur interne : 000284 ( Main/Exploration ); précédent : 000283; suivant : 000285Optimization of Agroinfiltration in Pisum sativum Provides a New Tool for Studying the Salivary Protein Functions in the Pea Aphid Complex.
Auteurs : Endrick Guy [France] ; Hélène Boulain [France] ; Yoann Aigu [France] ; Charlotte Le Pennec [France] ; Khaoula Chawki [France] ; Stéphanie Morlière [France] ; Kristina Sch Del [Allemagne] ; Grit Kunert [Allemagne] ; Jean-Christophe Simon [France] ; Akiko Sugio [France]Source :
- Frontiers in plant science [ 1664-462X ] ; 2016.
Abstract
Aphids are piercing-sucking insect pests and feed on phloem sap. During feeding, aphids inject a battery of salivary proteins into host plant. Some of these proteins function like effectors of microbial pathogens and influence the outcome of plant-aphid interactions. The pea aphid (Acyrthosiphon pisum) is the model aphid and encompasses multiple biotypes each specialized to one or a few legume species, providing an opportunity to investigate the underlying mechanisms of the compatibility between plants and aphid biotypes. We aim to identify the aphid factors that determine the compatibility with host plants, hence involved in the host plant specialization process, and hypothesize that salivary proteins are one of those factors. Agrobacterium-mediated transient gene expression is a powerful tool to perform functional analyses of effector (salivary) proteins in plants. However, the tool was not established for the legume species that A. pisum feeds on. Thus, we decided to optimize the method for legume plants to facilitate the functional analyses of A. pisum salivary proteins. We screened a range of cultivars of pea (Pisum sativum) and alfalfa (Medicago sativa). None of the M. sativa cultivars was suitable for agroinfiltration under the tested conditions; however, we established a protocol for efficient transient gene expression in two cultivars of P. sativum, ZP1109 and ZP1130, using A. tumefaciens AGL-1 strain and the pEAQ-HT-DEST1 vector. We confirmed that the genes are expressed from 3 to 10 days post-infiltration and that aphid lines of the pea adapted biotype fed and reproduced on these two cultivars while lines of alfalfa and clover biotypes did not. Thus, the pea biotype recognizes these two cultivars as typical pea plants. By using a combination of ZP1109 and an A. pisum line, we defined an agroinfiltration procedure to examine the effect of in planta expression of selected salivary proteins on A. pisum fitness and demonstrated that transient expression of one candidate salivary gene increased the fecundity of the aphids. This result confirms that the agroinfiltration can be used to perform functional analyses of salivary proteins in P. sativum and consequently to study the molecular mechanisms underlying host specialization in the pea aphid complex.
DOI: 10.3389/fpls.2016.01171
PubMed: 27555856
PubMed Central: PMC4977312
Affiliations:
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Rheu</wicri:noRegion></affiliation></author><author><name sortKey="Boulain, Helene" sort="Boulain, Helene" uniqKey="Boulain H" first="Hélène" last="Boulain">Hélène Boulain</name><affiliation wicri:level="1"><nlm:affiliation>INRA, UMR1349, Institute of Genetics, Environment and Plant Protection Le Rheu, France.</nlm:affiliation><country xml:lang="fr">France</country><wicri:regionArea>INRA, UMR1349, Institute of Genetics, Environment and Plant Protection Le Rheu</wicri:regionArea><wicri:noRegion>Environment and Plant Protection Le Rheu</wicri:noRegion><wicri:noRegion>Environment and Plant Protection Le Rheu</wicri:noRegion></affiliation></author><author><name sortKey="Aigu, Yoann" sort="Aigu, Yoann" uniqKey="Aigu Y" first="Yoann" last="Aigu">Yoann Aigu</name><affiliation wicri:level="1"><nlm:affiliation>INRA, UMR1349, Institute of Genetics, Environment and Plant Protection Le Rheu, France.</nlm:affiliation><country xml:lang="fr">France</country><wicri:regionArea>INRA, UMR1349, Institute 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Rheu</wicri:noRegion><wicri:noRegion>Environment and Plant Protection Le Rheu</wicri:noRegion></affiliation></author><author><name sortKey="Sch Del, Kristina" sort="Sch Del, Kristina" uniqKey="Sch Del K" first="Kristina" last="Sch Del">Kristina Sch Del</name><affiliation wicri:level="1"><nlm:affiliation>Department of Biochemistry, Max Planck Institute for Chemical Ecology Jena, Germany.</nlm:affiliation><country xml:lang="fr">Allemagne</country><wicri:regionArea>Department of Biochemistry, Max Planck Institute for Chemical Ecology Jena</wicri:regionArea><wicri:noRegion>Max Planck Institute for Chemical Ecology Jena</wicri:noRegion><wicri:noRegion>Max Planck Institute for Chemical Ecology Jena</wicri:noRegion><wicri:noRegion>Max Planck Institute for Chemical Ecology Jena</wicri:noRegion></affiliation></author><author><name sortKey="Kunert, Grit" sort="Kunert, Grit" uniqKey="Kunert G" first="Grit" last="Kunert">Grit Kunert</name><affiliation wicri:level="1"><nlm:affiliation>Department of Biochemistry, Max Planck Institute for Chemical Ecology Jena, Germany.</nlm:affiliation><country xml:lang="fr">Allemagne</country><wicri:regionArea>Department of Biochemistry, Max Planck Institute for Chemical Ecology Jena</wicri:regionArea><wicri:noRegion>Max Planck Institute for Chemical Ecology Jena</wicri:noRegion><wicri:noRegion>Max Planck Institute for Chemical Ecology Jena</wicri:noRegion><wicri:noRegion>Max Planck Institute for Chemical Ecology Jena</wicri:noRegion></affiliation></author><author><name sortKey="Simon, Jean Christophe" sort="Simon, Jean Christophe" uniqKey="Simon J" first="Jean-Christophe" last="Simon">Jean-Christophe Simon</name><affiliation wicri:level="1"><nlm:affiliation>INRA, UMR1349, Institute of Genetics, Environment and Plant Protection Le Rheu, France.</nlm:affiliation><country xml:lang="fr">France</country><wicri:regionArea>INRA, UMR1349, Institute of Genetics, Environment and Plant Protection Le Rheu</wicri:regionArea><wicri:noRegion>Environment and Plant Protection Le Rheu</wicri:noRegion><wicri:noRegion>Environment and Plant Protection Le Rheu</wicri:noRegion></affiliation></author><author><name sortKey="Sugio, Akiko" sort="Sugio, Akiko" uniqKey="Sugio A" first="Akiko" last="Sugio">Akiko Sugio</name><affiliation wicri:level="1"><nlm:affiliation>INRA, UMR1349, Institute of Genetics, Environment and Plant Protection Le Rheu, France.</nlm:affiliation><country xml:lang="fr">France</country><wicri:regionArea>INRA, UMR1349, Institute of Genetics, Environment and Plant Protection Le Rheu</wicri:regionArea><wicri:noRegion>Environment and Plant Protection Le Rheu</wicri:noRegion><wicri:noRegion>Environment and Plant Protection Le Rheu</wicri:noRegion></affiliation></author></analytic><series><title level="j">Frontiers in plant science</title><idno type="ISSN">1664-462X</idno><imprint><date when="2016" type="published">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Aphids are piercing-sucking insect pests and feed on phloem sap. During feeding, aphids inject a battery of salivary proteins into host plant. Some of these proteins function like effectors of microbial pathogens and influence the outcome of plant-aphid interactions. The pea aphid (Acyrthosiphon pisum) is the model aphid and encompasses multiple biotypes each specialized to one or a few legume species, providing an opportunity to investigate the underlying mechanisms of the compatibility between plants and aphid biotypes. We aim to identify the aphid factors that determine the compatibility with host plants, hence involved in the host plant specialization process, and hypothesize that salivary proteins are one of those factors. Agrobacterium-mediated transient gene expression is a powerful tool to perform functional analyses of effector (salivary) proteins in plants. However, the tool was not established for the legume species that A. pisum feeds on. Thus, we decided to optimize the method for legume plants to facilitate the functional analyses of A. pisum salivary proteins. We screened a range of cultivars of pea (Pisum sativum) and alfalfa (Medicago sativa). None of the M. sativa cultivars was suitable for agroinfiltration under the tested conditions; however, we established a protocol for efficient transient gene expression in two cultivars of P. sativum, ZP1109 and ZP1130, using A. tumefaciens AGL-1 strain and the pEAQ-HT-DEST1 vector. We confirmed that the genes are expressed from 3 to 10 days post-infiltration and that aphid lines of the pea adapted biotype fed and reproduced on these two cultivars while lines of alfalfa and clover biotypes did not. Thus, the pea biotype recognizes these two cultivars as typical pea plants. By using a combination of ZP1109 and an A. pisum line, we defined an agroinfiltration procedure to examine the effect of in planta expression of selected salivary proteins on A. pisum fitness and demonstrated that transient expression of one candidate salivary gene increased the fecundity of the aphids. This result confirms that the agroinfiltration can be used to perform functional analyses of salivary proteins in P. sativum and consequently to study the molecular mechanisms underlying host specialization in the pea aphid complex. </div></front></TEI><pubmed><MedlineCitation Status="PubMed-not-MEDLINE" Owner="NLM"><PMID Version="1">27555856</PMID><DateCompleted><Year>2016</Year><Month>08</Month><Day>24</Day></DateCompleted><DateRevised><Year>2020</Year><Month>09</Month><Day>30</Day></DateRevised><Article PubModel="Electronic-eCollection"><Journal><ISSN IssnType="Print">1664-462X</ISSN><JournalIssue CitedMedium="Print"><Volume>7</Volume><PubDate><Year>2016</Year></PubDate></JournalIssue><Title>Frontiers in plant science</Title><ISOAbbreviation>Front Plant Sci</ISOAbbreviation></Journal><ArticleTitle>Optimization of Agroinfiltration in Pisum sativum Provides a New Tool for Studying the Salivary Protein Functions in the Pea Aphid Complex.</ArticleTitle><Pagination><MedlinePgn>1171</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.3389/fpls.2016.01171</ELocationID><Abstract><AbstractText>Aphids are piercing-sucking insect pests and feed on phloem sap. During feeding, aphids inject a battery of salivary proteins into host plant. Some of these proteins function like effectors of microbial pathogens and influence the outcome of plant-aphid interactions. The pea aphid (Acyrthosiphon pisum) is the model aphid and encompasses multiple biotypes each specialized to one or a few legume species, providing an opportunity to investigate the underlying mechanisms of the compatibility between plants and aphid biotypes. We aim to identify the aphid factors that determine the compatibility with host plants, hence involved in the host plant specialization process, and hypothesize that salivary proteins are one of those factors. Agrobacterium-mediated transient gene expression is a powerful tool to perform functional analyses of effector (salivary) proteins in plants. However, the tool was not established for the legume species that A. pisum feeds on. Thus, we decided to optimize the method for legume plants to facilitate the functional analyses of A. pisum salivary proteins. We screened a range of cultivars of pea (Pisum sativum) and alfalfa (Medicago sativa). None of the M. sativa cultivars was suitable for agroinfiltration under the tested conditions; however, we established a protocol for efficient transient gene expression in two cultivars of P. sativum, ZP1109 and ZP1130, using A. tumefaciens AGL-1 strain and the pEAQ-HT-DEST1 vector. We confirmed that the genes are expressed from 3 to 10 days post-infiltration and that aphid lines of the pea adapted biotype fed and reproduced on these two cultivars while lines of alfalfa and clover biotypes did not. Thus, the pea biotype recognizes these two cultivars as typical pea plants. By using a combination of ZP1109 and an A. pisum line, we defined an agroinfiltration procedure to examine the effect of in planta expression of selected salivary proteins on A. pisum fitness and demonstrated that transient expression of one candidate salivary gene increased the fecundity of the aphids. This result confirms that the agroinfiltration can be used to perform functional analyses of salivary proteins in P. sativum and consequently to study the molecular mechanisms underlying host specialization in the pea aphid complex. </AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Guy</LastName><ForeName>Endrick</ForeName><Initials>E</Initials><AffiliationInfo><Affiliation>INRA, UMR1349, Institute of Genetics, Environment and Plant Protection Le Rheu, France.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Boulain</LastName><ForeName>Hélène</ForeName><Initials>H</Initials><AffiliationInfo><Affiliation>INRA, UMR1349, Institute of Genetics, Environment and Plant Protection Le Rheu, France.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Aigu</LastName><ForeName>Yoann</ForeName><Initials>Y</Initials><AffiliationInfo><Affiliation>INRA, UMR1349, Institute of Genetics, Environment and Plant Protection Le Rheu, France.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Le Pennec</LastName><ForeName>Charlotte</ForeName><Initials>C</Initials><AffiliationInfo><Affiliation>INRA, UMR1349, Institute of Genetics, Environment and Plant Protection Le Rheu, France.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Chawki</LastName><ForeName>Khaoula</ForeName><Initials>K</Initials><AffiliationInfo><Affiliation>INRA, UMR1349, Institute of Genetics, Environment and Plant Protection Le Rheu, France.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Morlière</LastName><ForeName>Stéphanie</ForeName><Initials>S</Initials><AffiliationInfo><Affiliation>INRA, UMR1349, Institute of Genetics, Environment and Plant Protection Le Rheu, France.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Schädel</LastName><ForeName>Kristina</ForeName><Initials>K</Initials><AffiliationInfo><Affiliation>Department of Biochemistry, Max Planck Institute for Chemical Ecology Jena, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Kunert</LastName><ForeName>Grit</ForeName><Initials>G</Initials><AffiliationInfo><Affiliation>Department of Biochemistry, Max Planck Institute for Chemical Ecology Jena, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Simon</LastName><ForeName>Jean-Christophe</ForeName><Initials>JC</Initials><AffiliationInfo><Affiliation>INRA, UMR1349, Institute of Genetics, Environment and Plant Protection Le Rheu, France.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Sugio</LastName><ForeName>Akiko</ForeName><Initials>A</Initials><AffiliationInfo><Affiliation>INRA, UMR1349, Institute of Genetics, Environment and Plant Protection Le Rheu, France.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2016</Year><Month>08</Month><Day>09</Day></ArticleDate></Article><MedlineJournalInfo><Country>Switzerland</Country><MedlineTA>Front Plant Sci</MedlineTA><NlmUniqueID>101568200</NlmUniqueID><ISSNLinking>1664-462X</ISSNLinking></MedlineJournalInfo><CommentsCorrectionsList><CommentsCorrections RefType="ErratumIn"><RefSource>Front Plant Sci. 2017 Jan 10;7:2046</RefSource><PMID Version="1">28077941</PMID></CommentsCorrections></CommentsCorrectionsList><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">Acyrthosiphon pisum</Keyword><Keyword MajorTopicYN="N">Leguminosae</Keyword><Keyword MajorTopicYN="N">agroinfiltration</Keyword><Keyword MajorTopicYN="N">biotypes</Keyword><Keyword MajorTopicYN="N">effector</Keyword><Keyword MajorTopicYN="N">host specialization</Keyword><Keyword MajorTopicYN="N">pea aphid</Keyword><Keyword MajorTopicYN="N">salivary proteins</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate 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