Efficient Agrobacterium-mediated transformation of commercial hybrid poplar Populus nigra L. x P. maximowiczii A. Henry.
Identifieur interne : 003286 ( Main/Exploration ); précédent : 003285; suivant : 003287Efficient Agrobacterium-mediated transformation of commercial hybrid poplar Populus nigra L. x P. maximowiczii A. Henry.
Auteurs : Dmytro P. Yevtushenko [Canada] ; Santosh MisraSource :
- Plant cell reports [ 1432-203X ] ; 2010.
Descripteurs français
- KwdFr :
- Milieux de culture (MeSH), Phénylurées (MeSH), Populus (génétique), Pousses de plante (génétique), Rhizobium (génétique), Régénération (MeSH), Techniques de transfert de gènes (MeSH), Thiadiazoles (MeSH), Transformation génétique (MeSH), Transgènes (MeSH), Vecteurs génétiques (MeSH), Végétaux génétiquement modifiés (génétique).
- MESH :
English descriptors
- KwdEn :
- Culture Media (MeSH), Gene Transfer Techniques (MeSH), Genetic Vectors (MeSH), Phenylurea Compounds (MeSH), Plant Shoots (genetics), Plants, Genetically Modified (genetics), Populus (genetics), Regeneration (MeSH), Rhizobium (genetics), Thiadiazoles (MeSH), Transformation, Genetic (MeSH), Transgenes (MeSH).
- MESH :
Abstract
Many economically important species of Populus, especially those in sections Aigeiros and Tacamahaca, remain recalcitrant to genetic transformation. In this study, a simple and reliable protocol was developed for the efficient Agrobacterium-mediated transformation of a difficult-to-transform, but commercially viable, hybrid poplar Populus nigra L. x P. maximowiczii A. Henry (NM6). A plant transformation vector designed to express the beta-glucuronidase (GUS) gene was used to detect transformation events at early stages of plant regeneration and to optimize parameters affecting poplar transformation. The use of zeatin riboside in shoot-induction medium, regeneration of shoots via indirect organogenesis, and early selection pressure were the major modifications that drastically improved the efficiency of poplar transformation and minimized the number of untransformed regenerants. Transgenic shoots were routinely obtained 4-10 weeks after co-culture with A. tumefaciens, with a greater than 90% rate of plant recovery. Stable transgene integration, ranging from a single insertion to ten copies per genome, was confirmed by Southern blot analysis. The mean transformation frequency was 36.3% and about two-thirds of the lines had 1-2 transgene copies. Among the explants, petioles and leaves had a higher transformation frequency than did stem segments. Growth characteristics and the morphology of transgenic poplar plants were identical to untransformed controls. These findings will accelerate the development of P. nigra x P. maximowiczii plants with novel traits, and may also be useful to improve transformation procedures for other Populus species.
DOI: 10.1007/s00299-009-0806-z
PubMed: 20087597
Affiliations:
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Yevtushenko</name><affiliation wicri:level="1"><nlm:affiliation>Department of Biochemistry and Microbiology, Centre for Forest Biology, University of Victoria, Victoria, BC, V8W 3P6, Canada. dmytro@uvic.ca</nlm:affiliation><country xml:lang="fr">Canada</country><wicri:regionArea>Department of Biochemistry and Microbiology, Centre for Forest Biology, University of Victoria, Victoria, BC, V8W 3P6</wicri:regionArea><wicri:noRegion>V8W 3P6</wicri:noRegion></affiliation></author><author><name sortKey="Misra, Santosh" sort="Misra, Santosh" uniqKey="Misra S" first="Santosh" last="Misra">Santosh Misra</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2010">2010</date><idno type="RBID">pubmed:20087597</idno><idno type="pmid">20087597</idno><idno type="doi">10.1007/s00299-009-0806-z</idno><idno type="wicri:Area/Main/Corpus">003327</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">003327</idno><idno type="wicri:Area/Main/Curation">003327</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Curation">003327</idno><idno type="wicri:Area/Main/Exploration">003327</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Efficient Agrobacterium-mediated transformation of commercial hybrid poplar Populus nigra L. x P. maximowiczii A. 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In this study, a simple and reliable protocol was developed for the efficient Agrobacterium-mediated transformation of a difficult-to-transform, but commercially viable, hybrid poplar Populus nigra L. x P. maximowiczii A. Henry (NM6). A plant transformation vector designed to express the beta-glucuronidase (GUS) gene was used to detect transformation events at early stages of plant regeneration and to optimize parameters affecting poplar transformation. The use of zeatin riboside in shoot-induction medium, regeneration of shoots via indirect organogenesis, and early selection pressure were the major modifications that drastically improved the efficiency of poplar transformation and minimized the number of untransformed regenerants. Transgenic shoots were routinely obtained 4-10 weeks after co-culture with A. tumefaciens, with a greater than 90% rate of plant recovery. Stable transgene integration, ranging from a single insertion to ten copies per genome, was confirmed by Southern blot analysis. The mean transformation frequency was 36.3% and about two-thirds of the lines had 1-2 transgene copies. Among the explants, petioles and leaves had a higher transformation frequency than did stem segments. Growth characteristics and the morphology of transgenic poplar plants were identical to untransformed controls. These findings will accelerate the development of P. nigra x P. maximowiczii plants with novel traits, and may also be useful to improve transformation procedures for other Populus species.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">20087597</PMID><DateCompleted><Year>2010</Year><Month>04</Month><Day>19</Day></DateCompleted><DateRevised><Year>2020</Year><Month>09</Month><Day>30</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1432-203X</ISSN><JournalIssue CitedMedium="Internet"><Volume>29</Volume><Issue>3</Issue><PubDate><Year>2010</Year><Month>Mar</Month></PubDate></JournalIssue><Title>Plant cell reports</Title><ISOAbbreviation>Plant Cell Rep</ISOAbbreviation></Journal><ArticleTitle>Efficient Agrobacterium-mediated transformation of commercial hybrid poplar Populus nigra L. x P. maximowiczii A. Henry.</ArticleTitle><Pagination><MedlinePgn>211-21</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1007/s00299-009-0806-z</ELocationID><Abstract><AbstractText>Many economically important species of Populus, especially those in sections Aigeiros and Tacamahaca, remain recalcitrant to genetic transformation. In this study, a simple and reliable protocol was developed for the efficient Agrobacterium-mediated transformation of a difficult-to-transform, but commercially viable, hybrid poplar Populus nigra L. x P. maximowiczii A. Henry (NM6). A plant transformation vector designed to express the beta-glucuronidase (GUS) gene was used to detect transformation events at early stages of plant regeneration and to optimize parameters affecting poplar transformation. The use of zeatin riboside in shoot-induction medium, regeneration of shoots via indirect organogenesis, and early selection pressure were the major modifications that drastically improved the efficiency of poplar transformation and minimized the number of untransformed regenerants. Transgenic shoots were routinely obtained 4-10 weeks after co-culture with A. tumefaciens, with a greater than 90% rate of plant recovery. Stable transgene integration, ranging from a single insertion to ten copies per genome, was confirmed by Southern blot analysis. The mean transformation frequency was 36.3% and about two-thirds of the lines had 1-2 transgene copies. Among the explants, petioles and leaves had a higher transformation frequency than did stem segments. Growth characteristics and the morphology of transgenic poplar plants were identical to untransformed controls. 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IdType="pubmed">17492451</ArticleId></ArticleIdList></Reference></ReferenceList></PubmedData></pubmed><affiliations><list><country><li>Canada</li></country></list><tree><noCountry><name sortKey="Misra, Santosh" sort="Misra, Santosh" uniqKey="Misra S" first="Santosh" last="Misra">Santosh Misra</name></noCountry><country name="Canada"><noRegion><name sortKey="Yevtushenko, Dmytro P" sort="Yevtushenko, Dmytro P" uniqKey="Yevtushenko D" first="Dmytro P" last="Yevtushenko">Dmytro P. 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