Repression of chilling-induced ACC accumulation in transgenic citrus by over-production of antisense 1-aminocyclopropane-1-carboxylate synthase RNA
Identifieur interne : 000638 ( Istex/Corpus ); précédent : 000637; suivant : 000639Repression of chilling-induced ACC accumulation in transgenic citrus by over-production of antisense 1-aminocyclopropane-1-carboxylate synthase RNA
Auteurs : Wai Shing Wong ; Geng Guang Li ; Wen Ning ; Zeng Fu Xu ; W. L. Wendy Hsiao ; Lang Yin Zhang ; Ning LiSource :
- Plant Science [ 0168-9452 ] ; 2001.
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Abstract
A chilling-inducible ACC synthase gene (CS-ACS1) has recently been identified from Citrus sinensis. This CS-ACS1 gene was constructed in an inverted orientation and placed under the control of the double 35S promoter. The antisense CS-ACS1 transgene was introduced into Carrizo citrange, C. sinensis (L.) Osbeck and Poncirus trifoliate by Agrobacterium-mediated gene transfer. The transformation efficiency in the transformation of citrus stem segment was improved significantly to 87 and 88% for Poncirus trifoliate (L.) Raf and Carrizo citrange, respectively. The transgenic citrus lines that produce higher level of antisense ACS RNA were found to repress the increase of ACC content following the chilling treatment. This work represents the first example in controlling the ethylene biosynthesis in citrus plants through the genetic engineering approach.
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DOI: 10.1016/S0168-9452(01)00505-2
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L. Wendy" last="Hsiao">W. L. Wendy Hsiao</name><affiliation><mods:affiliation>Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong, SAR</mods:affiliation></affiliation></author><author><name sortKey="Zhang, Lang Yin" sort="Zhang, Lang Yin" uniqKey="Zhang L" first="Lang Yin" last="Zhang">Lang Yin Zhang</name><affiliation><mods:affiliation>The South China Institute of Botany, Guangzhou, China</mods:affiliation></affiliation></author><author><name sortKey="Li, Ning" sort="Li, Ning" uniqKey="Li N" first="Ning" last="Li">Ning Li</name><affiliation><mods:affiliation>E-mail: boningli@ust.hk</mods:affiliation></affiliation><affiliation><mods:affiliation>Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong, SAR</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">Plant Science</title><title level="j" type="abbrev">PSL</title><idno type="ISSN">0168-9452</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="2001">2001</date><biblScope unit="volume">161</biblScope><biblScope unit="issue">5</biblScope><biblScope unit="page" from="969">969</biblScope><biblScope unit="page" to="977">977</biblScope></imprint><idno type="ISSN">0168-9452</idno></series><idno type="istex">937146832020EE3D64639B829A9FA8D36036A8CB</idno><idno type="DOI">10.1016/S0168-9452(01)00505-2</idno><idno type="PII">S0168-9452(01)00505-2</idno></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0168-9452</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>ACC synthase</term><term>Antisense</term><term>Carrizo citrange</term><term>Chilling</term><term>Poncirus</term><term>Transgenic citrus</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A chilling-inducible ACC synthase gene (CS-ACS1) has recently been identified from Citrus sinensis. This CS-ACS1 gene was constructed in an inverted orientation and placed under the control of the double 35S promoter. The antisense CS-ACS1 transgene was introduced into Carrizo citrange, C. sinensis (L.) Osbeck and Poncirus trifoliate by Agrobacterium-mediated gene transfer. The transformation efficiency in the transformation of citrus stem segment was improved significantly to 87 and 88% for Poncirus trifoliate (L.) Raf and Carrizo citrange, respectively. The transgenic citrus lines that produce higher level of antisense ACS RNA were found to repress the increase of ACC content following the chilling treatment. This work represents the first example in controlling the ethylene biosynthesis in citrus plants through the genetic engineering approach.</div></front></TEI><istex><corpusName>elsevier</corpusName><author><json:item><name>Wai Shing Wong</name><affiliations><json:string>Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong, SAR</json:string></affiliations></json:item><json:item><name>Geng Guang Li</name><affiliations><json:string>The South China Institute of Botany, Guangzhou, China</json:string></affiliations></json:item><json:item><name>Wen Ning</name><affiliations><json:string>Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong, SAR</json:string></affiliations></json:item><json:item><name>Zeng Fu Xu</name><affiliations><json:string>The Biotechnology Center, Zhongshan University, Guangzhou, China</json:string></affiliations></json:item><json:item><name>W.L.Wendy Hsiao</name><affiliations><json:string>Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong, SAR</json:string></affiliations></json:item><json:item><name>Lang Yin Zhang</name><affiliations><json:string>The South China Institute of Botany, Guangzhou, China</json:string></affiliations></json:item><json:item><name>Ning Li</name><affiliations><json:string>E-mail: boningli@ust.hk</json:string><json:string>Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong, SAR</json:string></affiliations></json:item></author><subject><json:item><lang><json:string>eng</json:string></lang><value>Transgenic citrus</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Poncirus</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Carrizo citrange</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Antisense</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>ACC synthase</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Chilling</value></json:item></subject><language><json:string>eng</json:string></language><originalGenre><json:string>Full-length article</json:string></originalGenre><abstract>A chilling-inducible ACC synthase gene (CS-ACS1) has recently been identified from Citrus sinensis. This CS-ACS1 gene was constructed in an inverted orientation and placed under the control of the double 35S promoter. The antisense CS-ACS1 transgene was introduced into Carrizo citrange, C. sinensis (L.) Osbeck and Poncirus trifoliate by Agrobacterium-mediated gene transfer. The transformation efficiency in the transformation of citrus stem segment was improved significantly to 87 and 88% for Poncirus trifoliate (L.) Raf and Carrizo citrange, respectively. The transgenic citrus lines that produce higher level of antisense ACS RNA were found to repress the increase of ACC content following the chilling treatment. This work represents the first example in controlling the ethylene biosynthesis in citrus plants through the genetic engineering approach.</abstract><qualityIndicators><score>6.464</score><pdfVersion>1.2</pdfVersion><pdfPageSize>555 x 762 pts</pdfPageSize><refBibsNative>true</refBibsNative><keywordCount>6</keywordCount><abstractCharCount>859</abstractCharCount><pdfWordCount>5465</pdfWordCount><pdfCharCount>33247</pdfCharCount><pdfPageCount>9</pdfPageCount><abstractWordCount>122</abstractWordCount></qualityIndicators><title>Repression of chilling-induced ACC accumulation in transgenic citrus by over-production of antisense 1-aminocyclopropane-1-carboxylate synthase RNA</title><pii><json:string>S0168-9452(01)00505-2</json:string></pii><genre><json:string>research-article</json:string></genre><host><volume>161</volume><pii><json:string>S0168-9452(00)X0118-5</json:string></pii><pages><last>977</last><first>969</first></pages><issn><json:string>0168-9452</json:string></issn><issue>5</issue><genre><json:string>journal</json:string></genre><language><json:string>unknown</json:string></language><title>Plant Science</title><publicationDate>2001</publicationDate></host><categories><wos><json:string>BIOCHEMISTRY & MOLECULAR BIOLOGY</json:string><json:string>PLANT SCIENCES</json:string></wos></categories><publicationDate>2001</publicationDate><copyrightDate>2001</copyrightDate><doi><json:string>10.1016/S0168-9452(01)00505-2</json:string></doi><id>937146832020EE3D64639B829A9FA8D36036A8CB</id><score>0.33294767</score><fulltext><json:item><original>true</original><mimetype>application/pdf</mimetype><extension>pdf</extension><uri>https://api.istex.fr/document/937146832020EE3D64639B829A9FA8D36036A8CB/fulltext/pdf</uri></json:item><json:item><original>false</original><mimetype>application/zip</mimetype><extension>zip</extension><uri>https://api.istex.fr/document/937146832020EE3D64639B829A9FA8D36036A8CB/fulltext/zip</uri></json:item><istex:fulltextTEI uri="https://api.istex.fr/document/937146832020EE3D64639B829A9FA8D36036A8CB/fulltext/tei"><teiHeader><fileDesc><titleStmt><title level="a" type="main" xml:lang="en">Repression of chilling-induced ACC accumulation in transgenic citrus by over-production of antisense 1-aminocyclopropane-1-carboxylate synthase RNA</title></titleStmt><publicationStmt><authority>ISTEX</authority><publisher>ELSEVIER</publisher><availability><p>ELSEVIER</p></availability><date>2001</date></publicationStmt><notesStmt><note type="content">Fig. 1: (A) The recombinant antisense ACS-containing binary plasmid. The antisense CS-ACS1 (Accession No.: AJ012551) cDNA fragment corresponding to nucleotides 275 to 1461 of CS-ACS1 was digested with EcoRI and HindIII and the resulting antisense CS-ACS1 gene expression cassette was inserted into the region between left boarder (LB) and right boarder (RB) of the binary vector pBI121 containing a kanamycin selectable marker. 35S, CaMV 35S promoter. NOS-ter, nopaline synthase terminator. NPTII, neomycin phosphotransferase II. (B) Regeneration of the transformed citrus plants. (i) Internodal segments co-cultured with Agrobacterium strain, EHA105:pBI121-aACS, followed by regeneration on a medium containing cefotaxime and kanamycin (see Section 2). (ii) Continued cultivation of shoots on medium containing kanamycin (see Section 2). (iii) Rooting of regenerated shoots was induced in medium containing NAA (for Poncirus trifoliata only). (iv) The pot-grown putative transgenic citrus plant (Carrizo citrange).</note><note type="content">Fig. 2: Molecular analysis of transgenic Carrizo citrange (Citrus sinensis (L.) Osb x Poncirus trifoliate (L.) Raf). Each lane contained 3–5 μg of citrus genomic DNA. The probe was 32P-labelled cauliflower mosaic virus (CaMV) 35S promoter fragment (0.8 kb) prepared by random primer labeling. Lane 1 and 2, the regenerated and untransformed citrus plants. The rest are transgenic citrus lines. Lane 3, line 56-8a; lane 4, line 57-4a; lane 5, line 57-5a; lane 6, line 63-2a; lane 7, line 67-2a; lane 8, line 72-2b; lane 9, line 73-1c; and lane 10, line 76-3a.</note><note type="content">Fig. 3: Northern blot analysis of transgenic citrus plants expressing antisense ACS genes. (A) The untransformed control plant (56-1c) and transgenic Carrizo citrange (67-2a, 73-1c and 76-3a) grown in pots. (B) Northern blot analysis of transgene expression in citrus plants. A nylon membrane blotted with 15 μg of total RNA per lane from both untransformed and transformed citrus plants was hybridized with 32P-labeled sense CS-ACS1 cDNA strand (see Section 2) prepared by asymmetric PCR using CS-ACS1 as the template and SPP1 as primer (see Section 2). The amount of RNA samples loaded into each lane was visualized by ethidium bromide-staining, as shown at the bottom of the panel B. rRNA, ribosomal RNA. Lane 1 and 2, the untransformed control plants; lane 3–10, the transgenic Carrizo citrange lines 56-8a, 57-4a, 57-5a, 63-2a, 67-2a, 72-2b, 73-1c and 76-3a, respectively. Lane 11 and 12 are the transgenic Poncirus line 20-2a and 20-2b, respectively.</note><note type="content">Fig. 4: Repression of ACC production in the transgenic citrus lines. The untransformed control plant is labeled as 56-1c. The transgenic citrus lines 72-2b, 67-2a and 73-1c are antisense ACS RNA over-producers. Line 57-5a is the transgenic citrus plant expressing relatively less antisense ACS RNA. The open and hatched bars represent the untreated and chilling-treated transgenic citrus plants, respectively.</note><note type="content">Table 1: Summary of regeneration and transformation of citrus stem segments</note></notesStmt><sourceDesc><biblStruct type="inbook"><analytic><title level="a" type="main" xml:lang="en">Repression of chilling-induced ACC accumulation in transgenic citrus by over-production of antisense 1-aminocyclopropane-1-carboxylate synthase RNA</title><author xml:id="author-1"><persName><forename type="first">Wai Shing</forename><surname>Wong</surname></persName><affiliation>Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong, SAR</affiliation></author><author xml:id="author-2"><persName><forename type="first">Geng Guang</forename><surname>Li</surname></persName><affiliation>The South China Institute of Botany, Guangzhou, China</affiliation></author><author xml:id="author-3"><persName><forename type="first">Wen</forename><surname>Ning</surname></persName><affiliation>Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong, SAR</affiliation></author><author xml:id="author-4"><persName><forename type="first">Zeng Fu</forename><surname>Xu</surname></persName><affiliation>The Biotechnology Center, Zhongshan University, Guangzhou, China</affiliation></author><author xml:id="author-5"><persName><forename type="first">W.L.Wendy</forename><surname>Hsiao</surname></persName><affiliation>Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong, SAR</affiliation></author><author xml:id="author-6"><persName><forename type="first">Lang Yin</forename><surname>Zhang</surname></persName><affiliation>The South China Institute of Botany, Guangzhou, China</affiliation></author><author xml:id="author-7"><persName><forename type="first">Ning</forename><surname>Li</surname></persName><email>boningli@ust.hk</email><note type="correspondence"><p>Corresponding author. Fax: +852-2358-7335</p></note><affiliation>Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong, SAR</affiliation></author></analytic><monogr><title level="j">Plant Science</title><title level="j" type="abbrev">PSL</title><idno type="pISSN">0168-9452</idno><idno type="PII">S0168-9452(00)X0118-5</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="2001"></date><biblScope unit="volume">161</biblScope><biblScope unit="issue">5</biblScope><biblScope unit="page" from="969">969</biblScope><biblScope unit="page" to="977">977</biblScope></imprint></monogr><idno type="istex">937146832020EE3D64639B829A9FA8D36036A8CB</idno><idno type="DOI">10.1016/S0168-9452(01)00505-2</idno><idno type="PII">S0168-9452(01)00505-2</idno></biblStruct></sourceDesc></fileDesc><profileDesc><creation><date>2001</date></creation><langUsage><language ident="en">en</language></langUsage><abstract xml:lang="en"><p>A chilling-inducible ACC synthase gene (CS-ACS1) has recently been identified from Citrus sinensis. This CS-ACS1 gene was constructed in an inverted orientation and placed under the control of the double 35S promoter. The antisense CS-ACS1 transgene was introduced into Carrizo citrange, C. sinensis (L.) Osbeck and Poncirus trifoliate by Agrobacterium-mediated gene transfer. The transformation efficiency in the transformation of citrus stem segment was improved significantly to 87 and 88% for Poncirus trifoliate (L.) Raf and Carrizo citrange, respectively. The transgenic citrus lines that produce higher level of antisense ACS RNA were found to repress the increase of ACC content following the chilling treatment. This work represents the first example in controlling the ethylene biosynthesis in citrus plants through the genetic engineering approach.</p></abstract><textClass xml:lang="en"><keywords scheme="keyword"><list><head>Keywords</head><item><term>Transgenic citrus</term></item><item><term>Poncirus</term></item><item><term>Carrizo citrange</term></item><item><term>Antisense</term></item><item><term>ACC synthase</term></item><item><term>Chilling</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="2001-07-07">Modified</change><change when="2001">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><original>false</original><mimetype>text/plain</mimetype><extension>txt</extension><uri>https://api.istex.fr/document/937146832020EE3D64639B829A9FA8D36036A8CB/fulltext/txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Elsevier, elements deleted: ce:floats; body; tail"><istex:xmlDeclaration>version="1.0" encoding="utf-8"</istex:xmlDeclaration><istex:docType PUBLIC="-//ES//DTD journal article DTD version 4.5.2//EN//XML" URI="art452.dtd" name="istex:docType"><istex:entity SYSTEM="gr1" NDATA="IMAGE" name="gr1"></istex:entity><istex:entity SYSTEM="gr2" NDATA="IMAGE" name="gr2"></istex:entity><istex:entity SYSTEM="gr3" NDATA="IMAGE" name="gr3"></istex:entity><istex:entity SYSTEM="gr4" NDATA="IMAGE" name="gr4"></istex:entity></istex:docType><istex:document><converted-article version="4.5.2" docsubtype="fla" xml:lang="en"><item-info><jid>PSL</jid><aid>5755</aid><ce:pii>S0168-9452(01)00505-2</ce:pii><ce:doi>10.1016/S0168-9452(01)00505-2</ce:doi><ce:copyright type="unknown" year="2001"></ce:copyright></item-info><head><ce:title>Repression of chilling-induced ACC accumulation in transgenic citrus by over-production of antisense 1-aminocyclopropane-1-carboxylate synthase RNA</ce:title><ce:author-group><ce:author><ce:given-name>Wai Shing</ce:given-name><ce:surname>Wong</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Geng Guang</ce:given-name><ce:surname>Li</ce:surname><ce:cross-ref refid="AFF2"><ce:sup>b</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Wen</ce:given-name><ce:surname>Ning</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Zeng Fu</ce:given-name><ce:surname>Xu</ce:surname><ce:cross-ref refid="AFF3"><ce:sup>c</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>W.L.Wendy</ce:given-name><ce:surname>Hsiao</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Lang Yin</ce:given-name><ce:surname>Zhang</ce:surname><ce:cross-ref refid="AFF2"><ce:sup>b</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Ning</ce:given-name><ce:surname>Li</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref><ce:cross-ref refid="COR1">*</ce:cross-ref><ce:e-address>boningli@ust.hk</ce:e-address></ce:author><ce:affiliation id="AFF1"><ce:label>a</ce:label><ce:textfn>Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong, SAR</ce:textfn></ce:affiliation><ce:affiliation id="AFF2"><ce:label>b</ce:label><ce:textfn>The South China Institute of Botany, Guangzhou, China</ce:textfn></ce:affiliation><ce:affiliation id="AFF3"><ce:label>c</ce:label><ce:textfn>The Biotechnology Center, Zhongshan University, Guangzhou, China</ce:textfn></ce:affiliation><ce:correspondence id="COR1"><ce:label>*</ce:label><ce:text>Corresponding author. Fax: +852-2358-7335</ce:text></ce:correspondence></ce:author-group><ce:date-received day="27" month="5" year="2001"></ce:date-received><ce:date-revised day="7" month="7" year="2001"></ce:date-revised><ce:date-accepted day="8" month="7" year="2001"></ce:date-accepted><ce:abstract><ce:section-title>Abstract</ce:section-title><ce:abstract-sec><ce:simple-para>A chilling-inducible ACC synthase gene (<ce:italic>CS-ACS1</ce:italic>) has recently been identified from <ce:italic>Citrus sinensis</ce:italic>. This <ce:italic>CS-ACS1</ce:italic> gene was constructed in an inverted orientation and placed under the control of the double 35S promoter. The antisense CS-ACS1 transgene was introduced into <ce:italic>Carrizo citrange</ce:italic>, <ce:italic>C. sinensis</ce:italic> (L.) Osbeck and <ce:italic>Poncirus trifoliate</ce:italic> by <ce:italic>Agrobacterium</ce:italic>-mediated gene transfer. The transformation efficiency in the transformation of citrus stem segment was improved significantly to 87 and 88% for <ce:italic>Poncirus trifoliate</ce:italic> (L.) Raf and <ce:italic>Carrizo citrange</ce:italic>, respectively. The transgenic citrus lines that produce higher level of antisense ACS RNA were found to repress the increase of ACC content following the chilling treatment. This work represents the first example in controlling the ethylene biosynthesis in citrus plants through the genetic engineering approach.</ce:simple-para></ce:abstract-sec></ce:abstract><ce:keywords class="keyword"><ce:section-title>Keywords</ce:section-title><ce:keyword><ce:text>Transgenic citrus</ce:text></ce:keyword><ce:keyword><ce:text><ce:italic>Poncirus</ce:italic></ce:text></ce:keyword><ce:keyword><ce:text><ce:italic>Carrizo citrange</ce:italic></ce:text></ce:keyword><ce:keyword><ce:text>Antisense</ce:text></ce:keyword><ce:keyword><ce:text>ACC synthase</ce:text></ce:keyword><ce:keyword><ce:text>Chilling</ce:text></ce:keyword></ce:keywords></head></converted-article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo lang="en"><title>Repression of chilling-induced ACC accumulation in transgenic citrus by over-production of antisense 1-aminocyclopropane-1-carboxylate synthase RNA</title></titleInfo><titleInfo type="alternative" lang="en" contentType="CDATA"><title>Repression of chilling-induced ACC accumulation in transgenic citrus by over-production of antisense 1-aminocyclopropane-1-carboxylate synthase RNA</title></titleInfo><name type="personal"><namePart type="given">Wai Shing</namePart><namePart type="family">Wong</namePart><affiliation>Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong, SAR</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Geng Guang</namePart><namePart type="family">Li</namePart><affiliation>The South China Institute of Botany, Guangzhou, China</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Wen</namePart><namePart type="family">Ning</namePart><affiliation>Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong, SAR</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Zeng Fu</namePart><namePart type="family">Xu</namePart><affiliation>The Biotechnology Center, Zhongshan University, Guangzhou, China</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">W.L.Wendy</namePart><namePart type="family">Hsiao</namePart><affiliation>Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong, SAR</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Lang Yin</namePart><namePart type="family">Zhang</namePart><affiliation>The South China Institute of Botany, Guangzhou, China</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Ning</namePart><namePart type="family">Li</namePart><affiliation>E-mail: boningli@ust.hk</affiliation><affiliation>Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong, SAR</affiliation><description>Corresponding author. Fax: +852-2358-7335</description><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="research-article" displayLabel="Full-length article"></genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">2001</dateIssued><dateModified encoding="w3cdtf">2001-07-07</dateModified><copyrightDate encoding="w3cdtf">2001</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><physicalDescription><internetMediaType>text/html</internetMediaType></physicalDescription><abstract lang="en">A chilling-inducible ACC synthase gene (CS-ACS1) has recently been identified from Citrus sinensis. This CS-ACS1 gene was constructed in an inverted orientation and placed under the control of the double 35S promoter. The antisense CS-ACS1 transgene was introduced into Carrizo citrange, C. sinensis (L.) Osbeck and Poncirus trifoliate by Agrobacterium-mediated gene transfer. The transformation efficiency in the transformation of citrus stem segment was improved significantly to 87 and 88% for Poncirus trifoliate (L.) Raf and Carrizo citrange, respectively. The transgenic citrus lines that produce higher level of antisense ACS RNA were found to repress the increase of ACC content following the chilling treatment. This work represents the first example in controlling the ethylene biosynthesis in citrus plants through the genetic engineering approach.</abstract><note type="content">Fig. 1: (A) The recombinant antisense ACS-containing binary plasmid. The antisense CS-ACS1 (Accession No.: AJ012551) cDNA fragment corresponding to nucleotides 275 to 1461 of CS-ACS1 was digested with EcoRI and HindIII and the resulting antisense CS-ACS1 gene expression cassette was inserted into the region between left boarder (LB) and right boarder (RB) of the binary vector pBI121 containing a kanamycin selectable marker. 35S, CaMV 35S promoter. NOS-ter, nopaline synthase terminator. NPTII, neomycin phosphotransferase II. (B) Regeneration of the transformed citrus plants. (i) Internodal segments co-cultured with Agrobacterium strain, EHA105:pBI121-aACS, followed by regeneration on a medium containing cefotaxime and kanamycin (see Section 2). (ii) Continued cultivation of shoots on medium containing kanamycin (see Section 2). (iii) Rooting of regenerated shoots was induced in medium containing NAA (for Poncirus trifoliata only). (iv) The pot-grown putative transgenic citrus plant (Carrizo citrange).</note><note type="content">Fig. 2: Molecular analysis of transgenic Carrizo citrange (Citrus sinensis (L.) Osb x Poncirus trifoliate (L.) Raf). Each lane contained 3–5 μg of citrus genomic DNA. The probe was 32P-labelled cauliflower mosaic virus (CaMV) 35S promoter fragment (0.8 kb) prepared by random primer labeling. Lane 1 and 2, the regenerated and untransformed citrus plants. The rest are transgenic citrus lines. Lane 3, line 56-8a; lane 4, line 57-4a; lane 5, line 57-5a; lane 6, line 63-2a; lane 7, line 67-2a; lane 8, line 72-2b; lane 9, line 73-1c; and lane 10, line 76-3a.</note><note type="content">Fig. 3: Northern blot analysis of transgenic citrus plants expressing antisense ACS genes. (A) The untransformed control plant (56-1c) and transgenic Carrizo citrange (67-2a, 73-1c and 76-3a) grown in pots. (B) Northern blot analysis of transgene expression in citrus plants. A nylon membrane blotted with 15 μg of total RNA per lane from both untransformed and transformed citrus plants was hybridized with 32P-labeled sense CS-ACS1 cDNA strand (see Section 2) prepared by asymmetric PCR using CS-ACS1 as the template and SPP1 as primer (see Section 2). The amount of RNA samples loaded into each lane was visualized by ethidium bromide-staining, as shown at the bottom of the panel B. rRNA, ribosomal RNA. Lane 1 and 2, the untransformed control plants; lane 3–10, the transgenic Carrizo citrange lines 56-8a, 57-4a, 57-5a, 63-2a, 67-2a, 72-2b, 73-1c and 76-3a, respectively. Lane 11 and 12 are the transgenic Poncirus line 20-2a and 20-2b, respectively.</note><note type="content">Fig. 4: Repression of ACC production in the transgenic citrus lines. The untransformed control plant is labeled as 56-1c. The transgenic citrus lines 72-2b, 67-2a and 73-1c are antisense ACS RNA over-producers. Line 57-5a is the transgenic citrus plant expressing relatively less antisense ACS RNA. The open and hatched bars represent the untreated and chilling-treated transgenic citrus plants, respectively.</note><note type="content">Table 1: Summary of regeneration and transformation of citrus stem segments</note><subject lang="en"><genre>Keywords</genre><topic>Transgenic citrus</topic><topic>Poncirus</topic><topic>Carrizo citrange</topic><topic>Antisense</topic><topic>ACC synthase</topic><topic>Chilling</topic></subject><relatedItem type="host"><titleInfo><title>Plant Science</title></titleInfo><titleInfo type="abbreviated"><title>PSL</title></titleInfo><genre type="journal">journal</genre><originInfo><dateIssued encoding="w3cdtf">200110</dateIssued></originInfo><identifier type="ISSN">0168-9452</identifier><identifier type="PII">S0168-9452(00)X0118-5</identifier><part><date>200110</date><detail type="volume"><number>161</number><caption>vol.</caption></detail><detail type="issue"><number>5</number><caption>no.</caption></detail><extent unit="issue pages"><start>825</start><end>1043</end></extent><extent unit="pages"><start>969</start><end>977</end></extent></part></relatedItem><identifier type="istex">937146832020EE3D64639B829A9FA8D36036A8CB</identifier><identifier type="DOI">10.1016/S0168-9452(01)00505-2</identifier><identifier type="PII">S0168-9452(01)00505-2</identifier><recordInfo><recordContentSource>ELSEVIER</recordContentSource></recordInfo></mods></metadata><enrichments><istex:catWosTEI uri="https://api.istex.fr/document/937146832020EE3D64639B829A9FA8D36036A8CB/enrichments/catWos"><teiHeader><profileDesc><textClass><classCode scheme="WOS">BIOCHEMISTRY & MOLECULAR BIOLOGY</classCode><classCode scheme="WOS">PLANT SCIENCES</classCode></textClass></profileDesc></teiHeader></istex:catWosTEI></enrichments><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
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