Protoplast transformation and regeneration of transgenic valencia sweet orange plants containing a juice quality-related pectin methylesterase gene
Identifieur interne : 002822 ( Main/Exploration ); précédent : 002821; suivant : 002823Protoplast transformation and regeneration of transgenic valencia sweet orange plants containing a juice quality-related pectin methylesterase gene
Auteurs : WENWU GUO [République populaire de Chine] ; YANXIN DUAN [République populaire de Chine] ; Oscar Olivares-Fuster [Espagne] ; ZHENCAI WU [États-Unis] ; Covadonga R. Arias [États-Unis] ; Jacqueline K. Burns [États-Unis] ; Jude W. Grosser [États-Unis]Source :
- Plant cell reports : (Print) [ 0721-7714 ] ; 2005.
Descripteurs français
- Pascal (Inist)
- Wicri :
- topic : Plante transgénique.
English descriptors
- KwdEn :
Abstract
Valencia orange [Citrus sinensis (L.) Osbeck] is the leading commercial citrus species in the world for processed juice products; however, the presence of thermostable pectin methylesterase (TSPME) reduces its juice quality. A long-term strategy of this work is to eliminate or greatly reduce TSPME activity in Valencia orange. Previous work resulted in the isolation of a putative TSPME gene, CsPME4, associated with a thermostable protein fraction of Valencia orange juice. To begin research designed to overexpress CsPME4 to verify the thermostability of the protein product and/or to downregulate the gene, a sense gene cassette containing a gene-specific sequence from a putative TSPME cDNA and the enhanced green fluorescent protein (GFP) as a selectable marker was constructed (M2.1). In the work reported here, M2.1 plasmid DNA was transformed (polyethylene glycol-mediated) into protoplasts isolated from an embryogenic suspension culture of Valencia somaclone line B6-68, in an effort to obtain transgenic Valencia lines. A vigorous transformed line was identified via GFP expression, physically separated from non-transformed tissue, and cultured on somatic embryogenesis induction medium. One transgenic proembryo expressing GFP was recovered and multiple shoots were regenerated. The recovery of multiple transgenic plants was expedited by in vitro grafting. Polymerase chain reaction analysis revealed the presence of the PME gene in transgenic plants, and subsequent Southern blot analysis confirmed the presence of the eGFP gene. These transgenic plants show normal growth and minor morphological variation. The thermostability of PME in these plants will be assessed after flowering and fruit set. This is the first successful transfer of a target fruit-quality gene by protoplast transformation with recovery of transgenic plants in citrus. This method of transformation has the advantage over Agrobacterium-mediated transformation in that it requires no antibiotic-resistance genes.
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Arias</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Citrus Research and Education Center, University of Florida, 700 Experiment Station Road</s1><s2>Lake Alfred, FI 33850</s2><s3>USA</s3><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Lake Alfred, FI 33850</wicri:noRegion></affiliation></author><author><name sortKey="Burns, Jacqueline K" sort="Burns, Jacqueline K" uniqKey="Burns J" first="Jacqueline K." last="Burns">Jacqueline K. Burns</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Citrus Research and Education Center, University of Florida, 700 Experiment Station Road</s1><s2>Lake Alfred, FI 33850</s2><s3>USA</s3><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Lake Alfred, FI 33850</wicri:noRegion></affiliation></author><author><name sortKey="Grosser, Jude W" sort="Grosser, Jude W" uniqKey="Grosser J" first="Jude W." last="Grosser">Jude W. Grosser</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Citrus Research and Education Center, University of Florida, 700 Experiment Station Road</s1><s2>Lake Alfred, FI 33850</s2><s3>USA</s3><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Lake Alfred, FI 33850</wicri:noRegion></affiliation></author></analytic><series><title level="j" type="main">Plant cell reports : (Print)</title><title level="j" type="abbreviated">Plant cell rep. : (Print)</title><idno type="ISSN">0721-7714</idno><imprint><date when="2005">2005</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Plant cell reports : (Print)</title><title level="j" type="abbreviated">Plant cell rep. : (Print)</title><idno type="ISSN">0721-7714</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Citrus sinensis</term><term>Gene</term><term>Genetic transformation</term><term>Green fluorescent protein</term><term>Orange juice</term><term>Pectinesterase</term><term>Protoplast</term><term>Quality</term><term>Regeneration</term><term>Transgenic plant</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Protoplaste</term><term>Régénération</term><term>Qualité</term><term>Pectinesterase</term><term>Gène</term><term>Citrus sinensis</term><term>Protéine fluorescente verte</term><term>Plante transgénique</term><term>Jus d'orange</term><term>Transformation génétique</term></keywords><keywords scheme="Wicri" type="topic" xml:lang="fr"><term>Plante transgénique</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Valencia orange [Citrus sinensis (L.) Osbeck] is the leading commercial citrus species in the world for processed juice products; however, the presence of thermostable pectin methylesterase (TSPME) reduces its juice quality. A long-term strategy of this work is to eliminate or greatly reduce TSPME activity in Valencia orange. Previous work resulted in the isolation of a putative TSPME gene, CsPME4, associated with a thermostable protein fraction of Valencia orange juice. To begin research designed to overexpress CsPME4 to verify the thermostability of the protein product and/or to downregulate the gene, a sense gene cassette containing a gene-specific sequence from a putative TSPME cDNA and the enhanced green fluorescent protein (GFP) as a selectable marker was constructed (M2.1). In the work reported here, M2.1 plasmid DNA was transformed (polyethylene glycol-mediated) into protoplasts isolated from an embryogenic suspension culture of Valencia somaclone line B6-68, in an effort to obtain transgenic Valencia lines. A vigorous transformed line was identified via GFP expression, physically separated from non-transformed tissue, and cultured on somatic embryogenesis induction medium. One transgenic proembryo expressing GFP was recovered and multiple shoots were regenerated. The recovery of multiple transgenic plants was expedited by in vitro grafting. Polymerase chain reaction analysis revealed the presence of the PME gene in transgenic plants, and subsequent Southern blot analysis confirmed the presence of the eGFP gene. These transgenic plants show normal growth and minor morphological variation. The thermostability of PME in these plants will be assessed after flowering and fruit set. This is the first successful transfer of a target fruit-quality gene by protoplast transformation with recovery of transgenic plants in citrus. This method of transformation has the advantage over Agrobacterium-mediated transformation in that it requires no antibiotic-resistance genes.</div></front></TEI><affiliations><list><country><li>Espagne</li><li>République populaire de Chine</li><li>États-Unis</li></country></list><tree><country name="République populaire de Chine"><noRegion><name sortKey="Wenwu Guo" sort="Wenwu Guo" uniqKey="Wenwu Guo" last="Wenwu Guo">WENWU GUO</name></noRegion><name sortKey="Yanxin Duan" sort="Yanxin Duan" uniqKey="Yanxin Duan" last="Yanxin Duan">YANXIN DUAN</name></country><country name="Espagne"><noRegion><name sortKey="Olivares Fuster, Oscar" sort="Olivares Fuster, Oscar" uniqKey="Olivares Fuster O" first="Oscar" last="Olivares-Fuster">Oscar Olivares-Fuster</name></noRegion></country><country name="États-Unis"><noRegion><name sortKey="Zhencai Wu" sort="Zhencai Wu" uniqKey="Zhencai Wu" last="Zhencai Wu">ZHENCAI WU</name></noRegion><name sortKey="Arias, Covadonga R" sort="Arias, Covadonga R" uniqKey="Arias C" first="Covadonga R." last="Arias">Covadonga R. Arias</name><name sortKey="Burns, Jacqueline K" sort="Burns, Jacqueline K" uniqKey="Burns J" first="Jacqueline K." last="Burns">Jacqueline K. Burns</name><name sortKey="Grosser, Jude W" sort="Grosser, Jude W" uniqKey="Grosser J" first="Jude W." last="Grosser">Jude W. Grosser</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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