Agrobacterium-derived cytokinin influences plastid morphology and starch accumulation in Nicotiana benthamiana during transient assays.
Identifieur interne : 000422 ( Main/Exploration ); précédent : 000421; suivant : 000423Agrobacterium-derived cytokinin influences plastid morphology and starch accumulation in Nicotiana benthamiana during transient assays.
Auteurs : Jessica L. Erickson ; Jörg Ziegler ; David Guevara ; Steffen Abel ; Ralf B. Klösgen ; Jaideep Mathur ; Steven J. Rothstein ; Martin H. Schattat [Allemagne]Source :
- BMC plant biology [ 1471-2229 ] ; 2014.
Descripteurs français
- KwdFr :
- Agrobacterium tumefaciens (génétique), Agrobacterium tumefaciens (métabolisme), Amidon (métabolisme), Cytokinine (pharmacologie), Dosage biologique (méthodes), Expression des gènes (effets des médicaments et des substances chimiques), Gènes bactériens (MeSH), Gènes rapporteurs (MeSH), Phénotype (MeSH), Plastes (effets des médicaments et des substances chimiques), Plastes (métabolisme), Tabac (effets des médicaments et des substances chimiques), Tabac (métabolisme), Transformation génétique (effets des médicaments et des substances chimiques), Vecteurs génétiques (métabolisme).
- MESH :
- effets des médicaments et des substances chimiques : Expression des gènes, Plastes, Tabac, Transformation génétique.
- génétique : Agrobacterium tumefaciens.
- métabolisme : Agrobacterium tumefaciens, Amidon, Plastes, Tabac, Vecteurs génétiques.
- méthodes : Dosage biologique.
- pharmacologie : Cytokinine.
- Gènes bactériens, Gènes rapporteurs, Phénotype.
English descriptors
- KwdEn :
- Agrobacterium tumefaciens (genetics), Agrobacterium tumefaciens (metabolism), Biological Assay (methods), Cytokinins (pharmacology), Gene Expression (drug effects), Genes, Bacterial (MeSH), Genes, Reporter (MeSH), Genetic Vectors (metabolism), Phenotype (MeSH), Plastids (drug effects), Plastids (metabolism), Starch (metabolism), Tobacco (drug effects), Tobacco (metabolism), Transformation, Genetic (drug effects).
- MESH :
- chemical , metabolism : Starch.
- chemical , pharmacology : Cytokinins.
- drug effects : Gene Expression, Plastids, Tobacco, Transformation, Genetic.
- genetics : Agrobacterium tumefaciens.
- metabolism : Agrobacterium tumefaciens, Genetic Vectors, Plastids, Tobacco.
- methods : Biological Assay.
- Genes, Bacterial, Genes, Reporter, Phenotype.
Abstract
BACKGROUND
Agrobacterium tumefaciens-based transient assays have become a common tool for answering questions related to protein localization and gene expression in a cellular context. The use of these assays assumes that the transiently transformed cells are observed under relatively authentic physiological conditions and maintain 'normal' sub-cellular behaviour. Although this premise is widely accepted, the question of whether cellular organization and organelle morphology is altered in Agrobacterium-infiltrated cells has not been examined in detail. The first indications of an altered sub-cellular environment came from our observation that a common laboratory strain, GV3101(pMP90), caused a drastic increase in stromule frequency. Stromules, or 'stroma-filled-tubules' emanate from the surface of plastids and are sensitive to a variety of biotic and abiotic stresses. Starting from this observation, the goal of our experiments was to further characterize the changes to the cell resulting from short-term bacterial infestation, and to identify the factor responsible for eliciting these changes.
RESULTS
Using a protocol typical of transient assays we evaluated the impact of GV3101(pMP90) infiltration on chloroplast behaviour and morphology in Nicotiana benthamiana. Our experiments confirmed that GV3101(pMP90) consistently induces stromules and alters plastid position relative to the nucleus. These effects were found to be the result of strain-dependant secretion of cytokinin and its accumulation in the plant tissue. Bacterial production of the hormone was found to be dependant on the presence of a trans-zeatin synthase gene (tzs) located on the Ti plasmid of GV3101(pMP90). Bacteria-derived cytokinins were also correlated with changes to both soluble sugar level and starch accumulation.
CONCLUSION
Although we have chosen to focus on how transient Agrobacterium infestation alters plastid based parameters, these changes to the morphology and position of a single organelle, combined with the measured increases in sugar and starch content, suggest global changes to cell physiology. This indicates that cells visualized during transient assays may not be as 'normal' as was previously assumed. Our results suggest that the impact of the bacteria can be minimized by choosing Agrobacterium strains devoid of the tzs gene, but that the alterations to sub-cellular organization and cell carbohydrate status cannot be completely avoided using this strategy.
DOI: 10.1186/1471-2229-14-127
PubMed: 24886417
PubMed Central: PMC4062310
Affiliations:
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Erickson</name></author><author><name sortKey="Ziegler, Jorg" sort="Ziegler, Jorg" uniqKey="Ziegler J" first="Jörg" last="Ziegler">Jörg Ziegler</name></author><author><name sortKey="Guevara, David" sort="Guevara, David" uniqKey="Guevara D" first="David" last="Guevara">David Guevara</name></author><author><name sortKey="Abel, Steffen" sort="Abel, Steffen" uniqKey="Abel S" first="Steffen" last="Abel">Steffen Abel</name></author><author><name sortKey="Klosgen, Ralf B" sort="Klosgen, Ralf B" uniqKey="Klosgen R" first="Ralf B" last="Klösgen">Ralf B. Klösgen</name></author><author><name sortKey="Mathur, Jaideep" sort="Mathur, Jaideep" uniqKey="Mathur J" first="Jaideep" last="Mathur">Jaideep Mathur</name></author><author><name sortKey="Rothstein, Steven J" sort="Rothstein, Steven J" uniqKey="Rothstein S" first="Steven J" last="Rothstein">Steven J. Rothstein</name></author><author><name sortKey="Schattat, Martin H" sort="Schattat, Martin H" uniqKey="Schattat M" first="Martin H" last="Schattat">Martin H. Schattat</name><affiliation wicri:level="1"><nlm:affiliation>Abteilung Pflanzen Physiologie, Institut für Biologie-Pflanzenphysiologie, Martin-Luther-Universität Halle-Wittenberg, Weinbergweg 10, Halle/Saale 06120, Germany. martin.schattat@pflanzenphys.uni-halle.de.</nlm:affiliation><country xml:lang="fr">Allemagne</country><wicri:regionArea>Abteilung Pflanzen Physiologie, Institut für Biologie-Pflanzenphysiologie, Martin-Luther-Universität Halle-Wittenberg, Weinbergweg 10, Halle/Saale 06120</wicri:regionArea><wicri:noRegion>06120</wicri:noRegion><wicri:noRegion>06120</wicri:noRegion><wicri:noRegion>Halle/Saale 06120</wicri:noRegion></affiliation></author></analytic><series><title level="j">BMC plant biology</title><idno type="eISSN">1471-2229</idno><imprint><date when="2014" type="published">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Agrobacterium tumefaciens (genetics)</term><term>Agrobacterium tumefaciens (metabolism)</term><term>Biological Assay (methods)</term><term>Cytokinins (pharmacology)</term><term>Gene Expression (drug effects)</term><term>Genes, Bacterial (MeSH)</term><term>Genes, Reporter (MeSH)</term><term>Genetic Vectors (metabolism)</term><term>Phenotype (MeSH)</term><term>Plastids (drug effects)</term><term>Plastids (metabolism)</term><term>Starch (metabolism)</term><term>Tobacco (drug effects)</term><term>Tobacco (metabolism)</term><term>Transformation, Genetic (drug effects)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Agrobacterium tumefaciens (génétique)</term><term>Agrobacterium tumefaciens (métabolisme)</term><term>Amidon (métabolisme)</term><term>Cytokinine (pharmacologie)</term><term>Dosage biologique (méthodes)</term><term>Expression des gènes (effets des médicaments et des substances chimiques)</term><term>Gènes bactériens (MeSH)</term><term>Gènes rapporteurs (MeSH)</term><term>Phénotype (MeSH)</term><term>Plastes (effets des médicaments et des substances chimiques)</term><term>Plastes (métabolisme)</term><term>Tabac (effets des médicaments et des substances chimiques)</term><term>Tabac (métabolisme)</term><term>Transformation génétique (effets des médicaments et des substances chimiques)</term><term>Vecteurs génétiques (métabolisme)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Starch</term></keywords><keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>Cytokinins</term></keywords><keywords scheme="MESH" qualifier="drug effects" xml:lang="en"><term>Gene Expression</term><term>Plastids</term><term>Tobacco</term><term>Transformation, Genetic</term></keywords><keywords scheme="MESH" qualifier="effets des médicaments et des substances chimiques" xml:lang="fr"><term>Expression des gènes</term><term>Plastes</term><term>Tabac</term><term>Transformation génétique</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Agrobacterium tumefaciens</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Agrobacterium tumefaciens</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Agrobacterium tumefaciens</term><term>Genetic Vectors</term><term>Plastids</term><term>Tobacco</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Biological Assay</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Agrobacterium tumefaciens</term><term>Amidon</term><term>Plastes</term><term>Tabac</term><term>Vecteurs génétiques</term></keywords><keywords scheme="MESH" qualifier="méthodes" xml:lang="fr"><term>Dosage biologique</term></keywords><keywords scheme="MESH" qualifier="pharmacologie" xml:lang="fr"><term>Cytokinine</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Genes, Bacterial</term><term>Genes, Reporter</term><term>Phenotype</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Gènes bactériens</term><term>Gènes rapporteurs</term><term>Phénotype</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p><b>BACKGROUND</b></p><p>Agrobacterium tumefaciens-based transient assays have become a common tool for answering questions related to protein localization and gene expression in a cellular context. The use of these assays assumes that the transiently transformed cells are observed under relatively authentic physiological conditions and maintain 'normal' sub-cellular behaviour. Although this premise is widely accepted, the question of whether cellular organization and organelle morphology is altered in Agrobacterium-infiltrated cells has not been examined in detail. The first indications of an altered sub-cellular environment came from our observation that a common laboratory strain, GV3101(pMP90), caused a drastic increase in stromule frequency. Stromules, or 'stroma-filled-tubules' emanate from the surface of plastids and are sensitive to a variety of biotic and abiotic stresses. Starting from this observation, the goal of our experiments was to further characterize the changes to the cell resulting from short-term bacterial infestation, and to identify the factor responsible for eliciting these changes.</p></div><div type="abstract" xml:lang="en"><p><b>RESULTS</b></p><p>Using a protocol typical of transient assays we evaluated the impact of GV3101(pMP90) infiltration on chloroplast behaviour and morphology in Nicotiana benthamiana. Our experiments confirmed that GV3101(pMP90) consistently induces stromules and alters plastid position relative to the nucleus. These effects were found to be the result of strain-dependant secretion of cytokinin and its accumulation in the plant tissue. Bacterial production of the hormone was found to be dependant on the presence of a trans-zeatin synthase gene (tzs) located on the Ti plasmid of GV3101(pMP90). Bacteria-derived cytokinins were also correlated with changes to both soluble sugar level and starch accumulation.</p></div><div type="abstract" xml:lang="en"><p><b>CONCLUSION</b></p><p>Although we have chosen to focus on how transient Agrobacterium infestation alters plastid based parameters, these changes to the morphology and position of a single organelle, combined with the measured increases in sugar and starch content, suggest global changes to cell physiology. This indicates that cells visualized during transient assays may not be as 'normal' as was previously assumed. Our results suggest that the impact of the bacteria can be minimized by choosing Agrobacterium strains devoid of the tzs gene, but that the alterations to sub-cellular organization and cell carbohydrate status cannot be completely avoided using this strategy.</p></div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">24886417</PMID><DateCompleted><Year>2015</Year><Month>01</Month><Day>16</Day></DateCompleted><DateRevised><Year>2018</Year><Month>11</Month><Day>13</Day></DateRevised><Article PubModel="Electronic"><Journal><ISSN IssnType="Electronic">1471-2229</ISSN><JournalIssue CitedMedium="Internet"><Volume>14</Volume><PubDate><Year>2014</Year><Month>May</Month><Day>09</Day></PubDate></JournalIssue><Title>BMC plant biology</Title><ISOAbbreviation>BMC Plant Biol</ISOAbbreviation></Journal><ArticleTitle>Agrobacterium-derived cytokinin influences plastid morphology and starch accumulation in Nicotiana benthamiana during transient assays.</ArticleTitle><Pagination><MedlinePgn>127</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1186/1471-2229-14-127</ELocationID><Abstract><AbstractText Label="BACKGROUND" NlmCategory="BACKGROUND">Agrobacterium tumefaciens-based transient assays have become a common tool for answering questions related to protein localization and gene expression in a cellular context. The use of these assays assumes that the transiently transformed cells are observed under relatively authentic physiological conditions and maintain 'normal' sub-cellular behaviour. Although this premise is widely accepted, the question of whether cellular organization and organelle morphology is altered in Agrobacterium-infiltrated cells has not been examined in detail. The first indications of an altered sub-cellular environment came from our observation that a common laboratory strain, GV3101(pMP90), caused a drastic increase in stromule frequency. Stromules, or 'stroma-filled-tubules' emanate from the surface of plastids and are sensitive to a variety of biotic and abiotic stresses. Starting from this observation, the goal of our experiments was to further characterize the changes to the cell resulting from short-term bacterial infestation, and to identify the factor responsible for eliciting these changes.</AbstractText><AbstractText Label="RESULTS" NlmCategory="RESULTS">Using a protocol typical of transient assays we evaluated the impact of GV3101(pMP90) infiltration on chloroplast behaviour and morphology in Nicotiana benthamiana. Our experiments confirmed that GV3101(pMP90) consistently induces stromules and alters plastid position relative to the nucleus. These effects were found to be the result of strain-dependant secretion of cytokinin and its accumulation in the plant tissue. Bacterial production of the hormone was found to be dependant on the presence of a trans-zeatin synthase gene (tzs) located on the Ti plasmid of GV3101(pMP90). Bacteria-derived cytokinins were also correlated with changes to both soluble sugar level and starch accumulation.</AbstractText><AbstractText Label="CONCLUSION" NlmCategory="CONCLUSIONS">Although we have chosen to focus on how transient Agrobacterium infestation alters plastid based parameters, these changes to the morphology and position of a single organelle, combined with the measured increases in sugar and starch content, suggest global changes to cell physiology. This indicates that cells visualized during transient assays may not be as 'normal' as was previously assumed. Our results suggest that the impact of the bacteria can be minimized by choosing Agrobacterium strains devoid of the tzs gene, but that the alterations to sub-cellular organization and cell carbohydrate status cannot be completely avoided using this strategy.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Erickson</LastName><ForeName>Jessica L</ForeName><Initials>JL</Initials></Author><Author ValidYN="Y"><LastName>Ziegler</LastName><ForeName>Jörg</ForeName><Initials>J</Initials></Author><Author ValidYN="Y"><LastName>Guevara</LastName><ForeName>David</ForeName><Initials>D</Initials></Author><Author ValidYN="Y"><LastName>Abel</LastName><ForeName>Steffen</ForeName><Initials>S</Initials></Author><Author ValidYN="Y"><LastName>Klösgen</LastName><ForeName>Ralf B</ForeName><Initials>RB</Initials></Author><Author ValidYN="Y"><LastName>Mathur</LastName><ForeName>Jaideep</ForeName><Initials>J</Initials></Author><Author ValidYN="Y"><LastName>Rothstein</LastName><ForeName>Steven J</ForeName><Initials>SJ</Initials></Author><Author ValidYN="Y"><LastName>Schattat</LastName><ForeName>Martin H</ForeName><Initials>MH</Initials><AffiliationInfo><Affiliation>Abteilung Pflanzen Physiologie, Institut für Biologie-Pflanzenphysiologie, Martin-Luther-Universität Halle-Wittenberg, Weinbergweg 10, Halle/Saale 06120, Germany. martin.schattat@pflanzenphys.uni-halle.de.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2014</Year><Month>05</Month><Day>09</Day></ArticleDate></Article><MedlineJournalInfo><Country>England</Country><MedlineTA>BMC Plant Biol</MedlineTA><NlmUniqueID>100967807</NlmUniqueID><ISSNLinking>1471-2229</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D003583">Cytokinins</NameOfSubstance></Chemical><Chemical><RegistryNumber>9005-25-8</RegistryNumber><NameOfSubstance UI="D013213">Starch</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D016960" MajorTopicYN="N">Agrobacterium tumefaciens</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D001681" MajorTopicYN="N">Biological Assay</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D003583" MajorTopicYN="N">Cytokinins</DescriptorName><QualifierName UI="Q000494" MajorTopicYN="Y">pharmacology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D015870" MajorTopicYN="N">Gene Expression</DescriptorName><QualifierName UI="Q000187" MajorTopicYN="N">drug effects</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D005798" MajorTopicYN="N">Genes, Bacterial</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D017930" MajorTopicYN="N">Genes, Reporter</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D005822" MajorTopicYN="N">Genetic Vectors</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010641" MajorTopicYN="N">Phenotype</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D018087" MajorTopicYN="N">Plastids</DescriptorName><QualifierName UI="Q000187" MajorTopicYN="N">drug effects</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D013213" MajorTopicYN="N">Starch</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D014026" MajorTopicYN="N">Tobacco</DescriptorName><QualifierName UI="Q000187" MajorTopicYN="N">drug effects</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D014170" MajorTopicYN="N">Transformation, Genetic</DescriptorName><QualifierName UI="Q000187" MajorTopicYN="N">drug effects</QualifierName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2013</Year><Month>11</Month><Day>26</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2014</Year><Month>04</Month><Day>24</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2014</Year><Month>6</Month><Day>3</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate 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uniqKey="Erickson J" first="Jessica L" last="Erickson">Jessica L. Erickson</name><name sortKey="Guevara, David" sort="Guevara, David" uniqKey="Guevara D" first="David" last="Guevara">David Guevara</name><name sortKey="Klosgen, Ralf B" sort="Klosgen, Ralf B" uniqKey="Klosgen R" first="Ralf B" last="Klösgen">Ralf B. Klösgen</name><name sortKey="Mathur, Jaideep" sort="Mathur, Jaideep" uniqKey="Mathur J" first="Jaideep" last="Mathur">Jaideep Mathur</name><name sortKey="Rothstein, Steven J" sort="Rothstein, Steven J" uniqKey="Rothstein S" first="Steven J" last="Rothstein">Steven J. Rothstein</name><name sortKey="Ziegler, Jorg" sort="Ziegler, Jorg" uniqKey="Ziegler J" first="Jörg" last="Ziegler">Jörg Ziegler</name></noCountry><country name="Allemagne"><noRegion><name sortKey="Schattat, Martin H" sort="Schattat, Martin H" uniqKey="Schattat M" first="Martin H" last="Schattat">Martin H. Schattat</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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