Low Agrobacterium tumefaciens inoculum levels and a long co-culture period lead to reduced plant defense responses and increase transgenic shoot production of sunflower (Helianthus annuus L.)
Identifieur interne : 002204 ( Ncbi/Merge ); précédent : 002203; suivant : 002205Low Agrobacterium tumefaciens inoculum levels and a long co-culture period lead to reduced plant defense responses and increase transgenic shoot production of sunflower (Helianthus annuus L.)
Auteurs : Zhifen Zhang [États-Unis] ; John J. Finer [États-Unis]Source :
- In Vitro Cellular & Developmental Biology [ 1054-5476 ] ; 2016.
Abstract
Url:
DOI: 10.1007/s11627-016-9774-5
PubMed: 27746666
PubMed Central: 5042984
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Low <italic>Agrobacterium tumefaciens</italic> inoculum levels and a long co-culture period lead to reduced plant defense responses and increase transgenic shoot production of sunflower (<italic>Helianthus annuus</italic> L.)</title><author><name sortKey="Zhang, Zhifen" sort="Zhang, Zhifen" uniqKey="Zhang Z" first="Zhifen" last="Zhang">Zhifen Zhang</name><affiliation wicri:level="2"><nlm:aff id="Aff1">Department of Horticulture and Crop Science, OARDC/The Ohio State University, 1680 Madison Avenue, Wooster, OH 44691 USA</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Ohio</region></placeName><wicri:cityArea>Department of Horticulture and Crop Science, OARDC/The Ohio State University, 1680 Madison Avenue, Wooster</wicri:cityArea></affiliation><affiliation wicri:level="2"><nlm:aff id="Aff2">Department of Horticulture, The University of Georgia Tifton Campus, Tifton, GA 31793 USA</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Géorgie (États-Unis)</region></placeName><wicri:cityArea>Department of Horticulture, The University of Georgia Tifton Campus, Tifton</wicri:cityArea></affiliation></author><author><name sortKey="Finer, John J" sort="Finer, John J" uniqKey="Finer J" first="John J." last="Finer">John J. Finer</name><affiliation wicri:level="2"><nlm:aff id="Aff1">Department of Horticulture and Crop Science, OARDC/The Ohio State University, 1680 Madison Avenue, Wooster, OH 44691 USA</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Ohio</region></placeName><wicri:cityArea>Department of Horticulture and Crop Science, OARDC/The Ohio State University, 1680 Madison Avenue, Wooster</wicri:cityArea></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">27746666</idno><idno type="pmc">5042984</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5042984</idno><idno type="RBID">PMC:5042984</idno><idno type="doi">10.1007/s11627-016-9774-5</idno><date when="2016">2016</date><idno type="wicri:Area/Pmc/Corpus">000670</idno><idno type="wicri:Area/Pmc/Curation">000669</idno><idno type="wicri:Area/Pmc/Checkpoint">000116</idno><idno type="wicri:Area/Ncbi/Merge">002204</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Low <italic>Agrobacterium tumefaciens</italic> inoculum levels and a long co-culture period lead to reduced plant defense responses and increase transgenic shoot production of sunflower (<italic>Helianthus annuus</italic> L.)</title><author><name sortKey="Zhang, Zhifen" sort="Zhang, Zhifen" uniqKey="Zhang Z" first="Zhifen" last="Zhang">Zhifen Zhang</name><affiliation wicri:level="2"><nlm:aff id="Aff1">Department of Horticulture and Crop Science, OARDC/The Ohio State University, 1680 Madison Avenue, Wooster, OH 44691 USA</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Ohio</region></placeName><wicri:cityArea>Department of Horticulture and Crop Science, OARDC/The Ohio State University, 1680 Madison Avenue, Wooster</wicri:cityArea></affiliation><affiliation wicri:level="2"><nlm:aff id="Aff2">Department of Horticulture, The University of Georgia Tifton Campus, Tifton, GA 31793 USA</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Géorgie (États-Unis)</region></placeName><wicri:cityArea>Department of Horticulture, The University of Georgia Tifton Campus, Tifton</wicri:cityArea></affiliation></author><author><name sortKey="Finer, John J" sort="Finer, John J" uniqKey="Finer J" first="John J." last="Finer">John J. Finer</name><affiliation wicri:level="2"><nlm:aff id="Aff1">Department of Horticulture and Crop Science, OARDC/The Ohio State University, 1680 Madison Avenue, Wooster, OH 44691 USA</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Ohio</region></placeName><wicri:cityArea>Department of Horticulture and Crop Science, OARDC/The Ohio State University, 1680 Madison Avenue, Wooster</wicri:cityArea></affiliation></author></analytic><series><title level="j">In Vitro Cellular & Developmental Biology</title><idno type="ISSN">1054-5476</idno><imprint><date when="2016">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p><italic>Agrobacterium</italic>-mediated plant transformation is typically conducted by inoculating plant tissues with an <italic>Agrobacterium</italic> suspension containing approximately 10<sup>8</sup>–10<sup>9</sup> bacteria mL<sup>−1</sup>, followed by a 2–3-d co-culture period. Use of longer co-culture periods could potentially increase transformation efficiencies by allowing more time for <italic>Agrobacterium</italic> to interact with plant cells, but bacterial overgrowth is likely to occur, leading to severe tissue browning and reduced transformation and regeneration. Low bacterial inoculum levels were therefore evaluated as a means to reduce the negative outcomes associated with long co-culture. The use of low inoculum bacterial suspensions (approximately 6 × 10<sup>2</sup> bacteria mL<sup>−1</sup>) followed by long co-culture (15 d) led to the production of an average of three transformed sunflower shoots per explant while the use of high inoculum (approximately 6 × 10<sup>8</sup> bacteria mL<sup>−1</sup>) followed by short co-culture (3 d) led to no transformed shoots. Low inoculum and long co-culture acted synergistically, and both were required for the improvement of sunflower transformation. Gene expression analysis via qRT-PCR showed that genes related to plant defense response were generally expressed at lower levels in the explants treated with low inoculum than those treated with high inoculum during 15 d of co-culture, suggesting that low inoculum reduced the induction of plant defense responses. The use of low inoculum with long co-culture (LI/LC) led to large increases in sunflower transformation efficiency. This method has great potential for improving transformation efficiencies and expanding the types of target tissues amenable for transformation of different plant species.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Alt Morbe, J" uniqKey="Alt Morbe J">J Alt-Mörbe</name></author><author><name sortKey="Kuhlmann, H" uniqKey="Kuhlmann H">H Kühlmann</name></author><author><name sortKey="Schroder, J" uniqKey="Schroder J">J Schröder</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Benzle, Ka" uniqKey="Benzle K">KA Benzle</name></author><author><name sortKey="Finer, Kr" uniqKey="Finer K">KR Finer</name></author><author><name sortKey="Marty, D" uniqKey="Marty D">D Marty</name></author><author><name sortKey="Mchale, Lk" uniqKey="Mchale L">LK McHale</name></author><author><name sortKey="Goodner, Bw" uniqKey="Goodner B">BW Goodner</name></author><author><name sortKey="Taylor, Cg" uniqKey="Taylor C">CG 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<front><journal-meta><journal-id journal-id-type="nlm-ta">In Vitro Cell Dev Biol Plant</journal-id><journal-id journal-id-type="iso-abbrev">In Vitro Cell. Dev. Biol., Plant</journal-id><journal-title-group><journal-title>In Vitro Cellular & Developmental Biology</journal-title></journal-title-group><issn pub-type="ppub">1054-5476</issn><publisher><publisher-name>Springer US</publisher-name><publisher-loc>New York</publisher-loc></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">27746666</article-id><article-id pub-id-type="pmc">5042984</article-id><article-id pub-id-type="publisher-id">9774</article-id><article-id pub-id-type="doi">10.1007/s11627-016-9774-5</article-id><article-categories><subj-group subj-group-type="heading"><subject>Genetic Transformation</subject></subj-group></article-categories><title-group><article-title>Low <italic>Agrobacterium tumefaciens</italic> inoculum levels and a long co-culture period lead to reduced plant defense responses and increase transgenic shoot production of sunflower (<italic>Helianthus annuus</italic> L.)</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Zhang</surname><given-names>Zhifen</given-names></name><xref ref-type="aff" rid="Aff1">1</xref><xref ref-type="aff" rid="Aff2">2</xref></contrib><contrib contrib-type="author" corresp="yes"><name><surname>Finer</surname><given-names>John J.</given-names></name><address><email>finer.1@osu.edu</email></address><xref ref-type="aff" rid="Aff1">1</xref></contrib><aff id="Aff1"><label>1</label>Department of Horticulture and Crop Science, OARDC/The Ohio State University, 1680 Madison Avenue, Wooster, OH 44691 USA</aff><aff id="Aff2"><label>2</label>Department of Horticulture, The University of Georgia Tifton Campus, Tifton, GA 31793 USA</aff></contrib-group><author-notes><fn fn-type="com"><p>Editor: David Duncan</p></fn></author-notes><pub-date pub-type="epub"><day>12</day><month>7</month><year>2016</year></pub-date><pub-date pub-type="pmc-release"><day>12</day><month>7</month><year>2016</year></pub-date><pub-date pub-type="ppub"><year>2016</year></pub-date><volume>52</volume><issue>4</issue><fpage>354</fpage><lpage>366</lpage><history><date date-type="received"><day>29</day><month>4</month><year>2016</year></date><date date-type="accepted"><day>28</day><month>6</month><year>2016</year></date></history><permissions><copyright-statement>© The Author(s) 2016</copyright-statement><license license-type="OpenAccess"><license-p><bold>Open Access</bold> This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.</license-p></license></permissions><abstract id="Abs1"><p><italic>Agrobacterium</italic>-mediated plant transformation is typically conducted by inoculating plant tissues with an <italic>Agrobacterium</italic> suspension containing approximately 10<sup>8</sup>–10<sup>9</sup> bacteria mL<sup>−1</sup>, followed by a 2–3-d co-culture period. Use of longer co-culture periods could potentially increase transformation efficiencies by allowing more time for <italic>Agrobacterium</italic> to interact with plant cells, but bacterial overgrowth is likely to occur, leading to severe tissue browning and reduced transformation and regeneration. Low bacterial inoculum levels were therefore evaluated as a means to reduce the negative outcomes associated with long co-culture. The use of low inoculum bacterial suspensions (approximately 6 × 10<sup>2</sup> bacteria mL<sup>−1</sup>) followed by long co-culture (15 d) led to the production of an average of three transformed sunflower shoots per explant while the use of high inoculum (approximately 6 × 10<sup>8</sup> bacteria mL<sup>−1</sup>) followed by short co-culture (3 d) led to no transformed shoots. Low inoculum and long co-culture acted synergistically, and both were required for the improvement of sunflower transformation. Gene expression analysis via qRT-PCR showed that genes related to plant defense response were generally expressed at lower levels in the explants treated with low inoculum than those treated with high inoculum during 15 d of co-culture, suggesting that low inoculum reduced the induction of plant defense responses. The use of low inoculum with long co-culture (LI/LC) led to large increases in sunflower transformation efficiency. This method has great potential for improving transformation efficiencies and expanding the types of target tissues amenable for transformation of different plant species.</p></abstract><kwd-group xml:lang="en"><title>Keywords</title><kwd><italic>Agrobacterium</italic></kwd><kwd>Inoculum density</kwd><kwd>Co-culture time</kwd><kwd>Shoot organogenesis</kwd><kwd>Plant defense</kwd></kwd-group><funding-group><award-group><funding-source><institution>USDA/Agricultural Research Service</institution></funding-source><award-id>58-5442-3-026</award-id></award-group></funding-group><custom-meta-group><custom-meta><meta-name>issue-copyright-statement</meta-name><meta-value>© The Society for In Vitro Biology 2016</meta-value></custom-meta></custom-meta-group></article-meta></front></pmc><affiliations><list><country><li>États-Unis</li></country><region><li>Géorgie (États-Unis)</li><li>Ohio</li></region></list><tree><country name="États-Unis"><region name="Ohio"><name sortKey="Zhang, Zhifen" sort="Zhang, Zhifen" uniqKey="Zhang Z" first="Zhifen" last="Zhang">Zhifen Zhang</name></region><name sortKey="Finer, John J" sort="Finer, John J" uniqKey="Finer J" first="John J." last="Finer">John J. Finer</name><name sortKey="Zhang, Zhifen" sort="Zhang, Zhifen" uniqKey="Zhang Z" first="Zhifen" last="Zhang">Zhifen Zhang</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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