Agrobacterium tumefaciens-mediated genetic transformation of a recalcitrant grain legume, lentil (Lens culinaris Medik).
Identifieur interne : 000660 ( Main/Exploration ); précédent : 000659; suivant : 000661Agrobacterium tumefaciens-mediated genetic transformation of a recalcitrant grain legume, lentil (Lens culinaris Medik).
Auteurs : Ufuk Celikkol Akcay [Turquie] ; M. Mahmoudian ; H. Kamci ; M. Yucel ; H A OktemSource :
- Plant cell reports [ 1432-203X ] ; 2009.
Descripteurs français
- KwdFr :
- MESH :
English descriptors
- KwdEn :
- MESH :
- chemical , genetics : DNA, Plant.
- chemical : DNA, Bacterial.
- genetics : Agrobacterium tumefaciens, Lens Plant, Plants, Genetically Modified.
- Gene Transfer Techniques, Transformation, Genetic, Transgenes.
Abstract
A simple and reproducible Agrobacterium-mediated transformation protocol for a recalcitrant legume plant, lentil (Lens culinaris M.) is reported. Application of wounding treatments and efficiencies of three Agrobacterium tumefaciens strains, EHA105, C58C1, and KYRT1 were compared for T-DNA delivery into lentil cotyledonary node tissues. KYRT1 was found to be on average 2.8-fold more efficient than both EHA105 and C58C1 for producing transient beta-glucuronidase (GUS) gene (gus) expression on cotyledonary petioles. Wounding of the explants, use of an optimized transformation protocol with the application of acetosyringone and vacuum infiltration treatments in addition to the application of a gradually intensifying selection regime played significant roles in enhancing transformation frequency. Lentil explants were transformed by inoculation with Agrobacterium tumefaciens strain, KYRT1 harboring a binary vector pTJK136 that carried neomycin phosphotransferase gene (npt-II) and an intron containing gusA gene on its T-DNA region. GUS-positive shoots were micrografted on lentil rootstocks. Transgenic lentil plants were produced with an overall transformation frequency of 2.3%. The presence of the transgene in the lentil genome was confirmed by GUS assay, PCR, RT-PCR and Southern hybridization. The transgenic shoots grafted on rootstocks were successfully transferred to soil and grown to maturity in the greenhouse. GUS activity was detected in vegetative and reproductive organs of T(0), T(1), T(2) and T(3) plants. PCR assays of T(1), T(2) and T(3) progenies confirmed the stable transmission of the transgene to the next generations.
DOI: 10.1007/s00299-008-0652-4
PubMed: 19083242
Affiliations:
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Mahmoudian</name></author><author><name sortKey="Kamci, H" sort="Kamci, H" uniqKey="Kamci H" first="H" last="Kamci">H. Kamci</name></author><author><name sortKey="Yucel, M" sort="Yucel, M" uniqKey="Yucel M" first="M" last="Yucel">M. Yucel</name></author><author><name sortKey="Oktem, H A" sort="Oktem, H A" uniqKey="Oktem H" first="H A" last="Oktem">H A Oktem</name></author></analytic><series><title level="j">Plant cell reports</title><idno type="eISSN">1432-203X</idno><imprint><date when="2009" type="published">2009</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Agrobacterium tumefaciens (genetics)</term><term>DNA, Bacterial (MeSH)</term><term>DNA, Plant (genetics)</term><term>Gene Transfer Techniques (MeSH)</term><term>Lens Plant (genetics)</term><term>Plants, Genetically Modified (genetics)</term><term>Transformation, Genetic (MeSH)</term><term>Transgenes (MeSH)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ADN bactérien (MeSH)</term><term>ADN des plantes (génétique)</term><term>Agrobacterium tumefaciens (génétique)</term><term>Lens (génétique)</term><term>Techniques de transfert de gènes (MeSH)</term><term>Transformation génétique (MeSH)</term><term>Transgènes (MeSH)</term><term>Végétaux génétiquement modifiés (génétique)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA, Plant</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>DNA, Bacterial</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Agrobacterium tumefaciens</term><term>Lens Plant</term><term>Plants, Genetically Modified</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ADN des plantes</term><term>Agrobacterium tumefaciens</term><term>Lens</term><term>Végétaux génétiquement modifiés</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Gene Transfer Techniques</term><term>Transformation, Genetic</term><term>Transgenes</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>ADN bactérien</term><term>Techniques de transfert de gènes</term><term>Transformation génétique</term><term>Transgènes</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A simple and reproducible Agrobacterium-mediated transformation protocol for a recalcitrant legume plant, lentil (Lens culinaris M.) is reported. Application of wounding treatments and efficiencies of three Agrobacterium tumefaciens strains, EHA105, C58C1, and KYRT1 were compared for T-DNA delivery into lentil cotyledonary node tissues. KYRT1 was found to be on average 2.8-fold more efficient than both EHA105 and C58C1 for producing transient beta-glucuronidase (GUS) gene (gus) expression on cotyledonary petioles. Wounding of the explants, use of an optimized transformation protocol with the application of acetosyringone and vacuum infiltration treatments in addition to the application of a gradually intensifying selection regime played significant roles in enhancing transformation frequency. Lentil explants were transformed by inoculation with Agrobacterium tumefaciens strain, KYRT1 harboring a binary vector pTJK136 that carried neomycin phosphotransferase gene (npt-II) and an intron containing gusA gene on its T-DNA region. GUS-positive shoots were micrografted on lentil rootstocks. Transgenic lentil plants were produced with an overall transformation frequency of 2.3%. The presence of the transgene in the lentil genome was confirmed by GUS assay, PCR, RT-PCR and Southern hybridization. The transgenic shoots grafted on rootstocks were successfully transferred to soil and grown to maturity in the greenhouse. GUS activity was detected in vegetative and reproductive organs of T(0), T(1), T(2) and T(3) plants. PCR assays of T(1), T(2) and T(3) progenies confirmed the stable transmission of the transgene to the next generations.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">19083242</PMID><DateCompleted><Year>2009</Year><Month>06</Month><Day>11</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>28</Volume><Issue>3</Issue><PubDate><Year>2009</Year><Month>Mar</Month></PubDate></JournalIssue><Title>Plant cell reports</Title><ISOAbbreviation>Plant Cell Rep</ISOAbbreviation></Journal><ArticleTitle>Agrobacterium tumefaciens-mediated genetic transformation of a recalcitrant grain legume, lentil (Lens culinaris Medik).</ArticleTitle><Pagination><MedlinePgn>407-17</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1007/s00299-008-0652-4</ELocationID><Abstract><AbstractText>A simple and reproducible Agrobacterium-mediated transformation protocol for a recalcitrant legume plant, lentil (Lens culinaris M.) is reported. Application of wounding treatments and efficiencies of three Agrobacterium tumefaciens strains, EHA105, C58C1, and KYRT1 were compared for T-DNA delivery into lentil cotyledonary node tissues. KYRT1 was found to be on average 2.8-fold more efficient than both EHA105 and C58C1 for producing transient beta-glucuronidase (GUS) gene (gus) expression on cotyledonary petioles. Wounding of the explants, use of an optimized transformation protocol with the application of acetosyringone and vacuum infiltration treatments in addition to the application of a gradually intensifying selection regime played significant roles in enhancing transformation frequency. Lentil explants were transformed by inoculation with Agrobacterium tumefaciens strain, KYRT1 harboring a binary vector pTJK136 that carried neomycin phosphotransferase gene (npt-II) and an intron containing gusA gene on its T-DNA region. GUS-positive shoots were micrografted on lentil rootstocks. Transgenic lentil plants were produced with an overall transformation frequency of 2.3%. The presence of the transgene in the lentil genome was confirmed by GUS assay, PCR, RT-PCR and Southern hybridization. The transgenic shoots grafted on rootstocks were successfully transferred to soil and grown to maturity in the greenhouse. GUS activity was detected in vegetative and reproductive organs of T(0), T(1), T(2) and T(3) plants. PCR assays of T(1), T(2) and T(3) progenies confirmed the stable transmission of the transgene to the next generations.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Akcay</LastName><ForeName>Ufuk Celikkol</ForeName><Initials>UC</Initials><AffiliationInfo><Affiliation>Department of Food Engineering, Suleyman Demirel University, Isparta, Turkey. ufukca@mmf.sdu.edu.tr</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Mahmoudian</LastName><ForeName>M</ForeName><Initials>M</Initials></Author><Author ValidYN="Y"><LastName>Kamci</LastName><ForeName>H</ForeName><Initials>H</Initials></Author><Author ValidYN="Y"><LastName>Yucel</LastName><ForeName>M</ForeName><Initials>M</Initials></Author><Author ValidYN="Y"><LastName>Oktem</LastName><ForeName>H A</ForeName><Initials>HA</Initials></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>2008</Year><Month>12</Month><Day>16</Day></ArticleDate></Article><MedlineJournalInfo><Country>Germany</Country><MedlineTA>Plant Cell Rep</MedlineTA><NlmUniqueID>9880970</NlmUniqueID><ISSNLinking>0721-7714</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D004269">DNA, Bacterial</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D018744">DNA, Plant</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="C036483">T-DNA</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D016960" MajorTopicYN="N">Agrobacterium tumefaciens</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName 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PubStatus="received"><Year>2008</Year><Month>09</Month><Day>04</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2008</Year><Month>11</Month><Day>23</Day></PubMedPubDate><PubMedPubDate PubStatus="revised"><Year>2008</Year><Month>10</Month><Day>16</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2008</Year><Month>12</Month><Day>17</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2008</Year><Month>12</Month><Day>17</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2009</Year><Month>6</Month><Day>12</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">19083242</ArticleId><ArticleId IdType="doi">10.1007/s00299-008-0652-4</ArticleId></ArticleIdList><ReferenceList><Reference><Citation>Plant Cell Rep. 2005 Jun;24(3):164-71</Citation><ArticleIdList><ArticleId 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Kamci</name><name sortKey="Mahmoudian, M" sort="Mahmoudian, M" uniqKey="Mahmoudian M" first="M" last="Mahmoudian">M. Mahmoudian</name><name sortKey="Oktem, H A" sort="Oktem, H A" uniqKey="Oktem H" first="H A" last="Oktem">H A Oktem</name><name sortKey="Yucel, M" sort="Yucel, M" uniqKey="Yucel M" first="M" last="Yucel">M. Yucel</name></noCountry><country name="Turquie"><noRegion><name sortKey="Akcay, Ufuk Celikkol" sort="Akcay, Ufuk Celikkol" uniqKey="Akcay U" first="Ufuk Celikkol" last="Akcay">Ufuk Celikkol Akcay</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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