Rapid, in situ detection of Agrobacterium tumefaciens attachment to leaf tissue.
Identifieur interne : 000490 ( Main/Exploration ); précédent : 000489; suivant : 000491Rapid, in situ detection of Agrobacterium tumefaciens attachment to leaf tissue.
Auteurs : Christopher W. Simmons [États-Unis] ; N. Nitin ; Jean S. VandergheynstSource :
- Biotechnology progress [ 1520-6033 ]
Descripteurs français
- KwdFr :
- Adhérence bactérienne (MeSH), Agrobacterium tumefaciens (composition chimique), Agrobacterium tumefaciens (physiologie), Colorants fluorescents (composition chimique), Coloration et marquage (méthodes), Feuilles de plante (composition chimique), Feuilles de plante (microbiologie), Laitue (composition chimique), Laitue (microbiologie).
- MESH :
- composition chimique : Agrobacterium tumefaciens, Colorants fluorescents, Feuilles de plante, Laitue.
- microbiologie : Feuilles de plante, Laitue.
- méthodes : Coloration et marquage.
- physiologie : Agrobacterium tumefaciens.
- Adhérence bactérienne.
English descriptors
- KwdEn :
- MESH :
- chemical , chemistry : Fluorescent Dyes.
- chemistry : Agrobacterium tumefaciens, Lettuce, Plant Leaves.
- methods : Staining and Labeling.
- microbiology : Lettuce, Plant Leaves.
- physiology : Agrobacterium tumefaciens.
- Bacterial Adhesion.
Abstract
Attachment of the plant pathogen Agrobacterium tumefaciens to host plant cells is an early and necessary step in plant transformation and agroinfiltration processes. However, bacterial attachment behavior is not well understood in complex plant tissues. Here we developed an imaging-based method to observe and quantify A. tumefaciens attached to leaf tissue in situ. Fluorescent labeling of bacteria with nucleic acid, protein, and vital dyes was investigated as a rapid alternative to generating recombinant strains expressing fluorescent proteins. Syto 16 green fluorescent nucleic acid stain was found to yield the greatest signal intensity in stained bacteria without affecting viability or infectivity. Stained bacteria retained the stain and were detectable over 72 h. To demonstrate in situ detection of attached bacteria, confocal fluorescent microscopy was used to image A. tumefaciens in sections of lettuce leaf tissue following vacuum-infiltration with labeled bacteria. Bacterial signals were associated with plant cell surfaces, suggesting detection of bacteria attached to plant cells. Bacterial attachment to specific leaf tissues was in agreement with known leaf tissue competencies for transformation with Agrobacterium. Levels of bacteria attached to leaf cells were quantified over time post-infiltration. Signals from stained bacteria were stable over the first 24 h following infiltration but decreased in intensity as bacteria multiplied in planta. Nucleic acid staining of A. tumefaciens followed by confocal microscopy of infected leaf tissue offers a rapid, in situ method for evaluating attachment of A. tumefaciens' to plant expression hosts and a tool to facilitate management of transient expression processes via agroinfiltration.
DOI: 10.1002/btpr.1608
PubMed: 22848046
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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Nitin</name></author><author><name sortKey="Vandergheynst, Jean S" sort="Vandergheynst, Jean S" uniqKey="Vandergheynst J" first="Jean S" last="Vandergheynst">Jean S. Vandergheynst</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2012">2012 Sep-Oct</date><idno type="RBID">pubmed:22848046</idno><idno type="pmid">22848046</idno><idno type="doi">10.1002/btpr.1608</idno><idno type="wicri:Area/Main/Corpus">000503</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">000503</idno><idno type="wicri:Area/Main/Curation">000503</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Curation">000503</idno><idno type="wicri:Area/Main/Exploration">000503</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Rapid, in situ detection of Agrobacterium tumefaciens attachment to leaf tissue.</title><author><name sortKey="Simmons, Christopher W" sort="Simmons, Christopher W" uniqKey="Simmons C" first="Christopher W" last="Simmons">Christopher W. 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Vandergheynst</name></author></analytic><series><title level="j">Biotechnology progress</title><idno type="eISSN">1520-6033</idno></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Agrobacterium tumefaciens (chemistry)</term><term>Agrobacterium tumefaciens (physiology)</term><term>Bacterial Adhesion (MeSH)</term><term>Fluorescent Dyes (chemistry)</term><term>Lettuce (chemistry)</term><term>Lettuce (microbiology)</term><term>Plant Leaves (chemistry)</term><term>Plant Leaves (microbiology)</term><term>Staining and Labeling (methods)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Adhérence bactérienne (MeSH)</term><term>Agrobacterium tumefaciens (composition chimique)</term><term>Agrobacterium tumefaciens (physiologie)</term><term>Colorants fluorescents (composition chimique)</term><term>Coloration et marquage (méthodes)</term><term>Feuilles de plante (composition chimique)</term><term>Feuilles de plante (microbiologie)</term><term>Laitue (composition chimique)</term><term>Laitue (microbiologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Fluorescent Dyes</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Agrobacterium tumefaciens</term><term>Lettuce</term><term>Plant Leaves</term></keywords><keywords scheme="MESH" qualifier="composition chimique" xml:lang="fr"><term>Agrobacterium tumefaciens</term><term>Colorants fluorescents</term><term>Feuilles de plante</term><term>Laitue</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Staining and Labeling</term></keywords><keywords scheme="MESH" qualifier="microbiologie" xml:lang="fr"><term>Feuilles de plante</term><term>Laitue</term></keywords><keywords scheme="MESH" qualifier="microbiology" xml:lang="en"><term>Lettuce</term><term>Plant Leaves</term></keywords><keywords scheme="MESH" qualifier="méthodes" xml:lang="fr"><term>Coloration et marquage</term></keywords><keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Agrobacterium tumefaciens</term></keywords><keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>Agrobacterium tumefaciens</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Bacterial Adhesion</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Adhérence bactérienne</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Attachment of the plant pathogen Agrobacterium tumefaciens to host plant cells is an early and necessary step in plant transformation and agroinfiltration processes. However, bacterial attachment behavior is not well understood in complex plant tissues. Here we developed an imaging-based method to observe and quantify A. tumefaciens attached to leaf tissue in situ. Fluorescent labeling of bacteria with nucleic acid, protein, and vital dyes was investigated as a rapid alternative to generating recombinant strains expressing fluorescent proteins. Syto 16 green fluorescent nucleic acid stain was found to yield the greatest signal intensity in stained bacteria without affecting viability or infectivity. Stained bacteria retained the stain and were detectable over 72 h. To demonstrate in situ detection of attached bacteria, confocal fluorescent microscopy was used to image A. tumefaciens in sections of lettuce leaf tissue following vacuum-infiltration with labeled bacteria. Bacterial signals were associated with plant cell surfaces, suggesting detection of bacteria attached to plant cells. Bacterial attachment to specific leaf tissues was in agreement with known leaf tissue competencies for transformation with Agrobacterium. Levels of bacteria attached to leaf cells were quantified over time post-infiltration. Signals from stained bacteria were stable over the first 24 h following infiltration but decreased in intensity as bacteria multiplied in planta. Nucleic acid staining of A. tumefaciens followed by confocal microscopy of infected leaf tissue offers a rapid, in situ method for evaluating attachment of A. tumefaciens' to plant expression hosts and a tool to facilitate management of transient expression processes via agroinfiltration.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">22848046</PMID><DateCompleted><Year>2013</Year><Month>03</Month><Day>01</Day></DateCompleted><DateRevised><Year>2019</Year><Month>12</Month><Day>10</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1520-6033</ISSN><JournalIssue CitedMedium="Internet"><Volume>28</Volume><Issue>5</Issue><PubDate><MedlineDate>2012 Sep-Oct</MedlineDate></PubDate></JournalIssue><Title>Biotechnology progress</Title><ISOAbbreviation>Biotechnol Prog</ISOAbbreviation></Journal><ArticleTitle>Rapid, in situ detection of Agrobacterium tumefaciens attachment to leaf tissue.</ArticleTitle><Pagination><MedlinePgn>1321-8</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1002/btpr.1608</ELocationID><Abstract><AbstractText>Attachment of the plant pathogen Agrobacterium tumefaciens to host plant cells is an early and necessary step in plant transformation and agroinfiltration processes. However, bacterial attachment behavior is not well understood in complex plant tissues. Here we developed an imaging-based method to observe and quantify A. tumefaciens attached to leaf tissue in situ. Fluorescent labeling of bacteria with nucleic acid, protein, and vital dyes was investigated as a rapid alternative to generating recombinant strains expressing fluorescent proteins. Syto 16 green fluorescent nucleic acid stain was found to yield the greatest signal intensity in stained bacteria without affecting viability or infectivity. Stained bacteria retained the stain and were detectable over 72 h. To demonstrate in situ detection of attached bacteria, confocal fluorescent microscopy was used to image A. tumefaciens in sections of lettuce leaf tissue following vacuum-infiltration with labeled bacteria. Bacterial signals were associated with plant cell surfaces, suggesting detection of bacteria attached to plant cells. Bacterial attachment to specific leaf tissues was in agreement with known leaf tissue competencies for transformation with Agrobacterium. Levels of bacteria attached to leaf cells were quantified over time post-infiltration. Signals from stained bacteria were stable over the first 24 h following infiltration but decreased in intensity as bacteria multiplied in planta. Nucleic acid staining of A. tumefaciens followed by confocal microscopy of infected leaf tissue offers a rapid, in situ method for evaluating attachment of A. tumefaciens' to plant expression hosts and a tool to facilitate management of transient expression processes via agroinfiltration.</AbstractText><CopyrightInformation>Copyright © 2012 American Institute of Chemical Engineers (AIChE).</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Simmons</LastName><ForeName>Christopher W</ForeName><Initials>CW</Initials><AffiliationInfo><Affiliation>Dept of Biological and Agricultural Engineering, University of California, Davis, One Shields Avenue, Davis, CA, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Nitin</LastName><ForeName>N</ForeName><Initials>N</Initials></Author><Author ValidYN="Y"><LastName>Vandergheynst</LastName><ForeName>Jean S</ForeName><Initials>JS</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D023362">Evaluation Study</PublicationType><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013486">Research Support, U.S. Gov't, Non-P.H.S.</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2012</Year><Month>08</Month><Day>28</Day></ArticleDate></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>Biotechnol Prog</MedlineTA><NlmUniqueID>8506292</NlmUniqueID><ISSNLinking>1520-6033</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D005456">Fluorescent Dyes</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D016960" MajorTopicYN="N">Agrobacterium tumefaciens</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="Y">chemistry</QualifierName><QualifierName UI="Q000502" MajorTopicYN="Y">physiology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D001422" MajorTopicYN="Y">Bacterial Adhesion</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D005456" MajorTopicYN="N">Fluorescent Dyes</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D018545" MajorTopicYN="N">Lettuce</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000382" MajorTopicYN="Y">microbiology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D018515" MajorTopicYN="N">Plant Leaves</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000382" MajorTopicYN="Y">microbiology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D013194" MajorTopicYN="N">Staining and Labeling</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2012</Year><Month>05</Month><Day>18</Day></PubMedPubDate><PubMedPubDate PubStatus="revised"><Year>2012</Year><Month>07</Month><Day>06</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2012</Year><Month>8</Month><Day>1</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2012</Year><Month>8</Month><Day>1</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2013</Year><Month>3</Month><Day>2</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">22848046</ArticleId><ArticleId IdType="doi">10.1002/btpr.1608</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>États-Unis</li></country><region><li>Californie</li></region></list><tree><noCountry><name sortKey="Nitin, N" sort="Nitin, N" uniqKey="Nitin N" first="N" last="Nitin">N. Nitin</name><name sortKey="Vandergheynst, Jean S" sort="Vandergheynst, Jean S" uniqKey="Vandergheynst J" first="Jean S" last="Vandergheynst">Jean S. Vandergheynst</name></noCountry><country name="États-Unis"><region name="Californie"><name sortKey="Simmons, Christopher W" sort="Simmons, Christopher W" uniqKey="Simmons C" first="Christopher W" last="Simmons">Christopher W. Simmons</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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