Use of the VvMybA1 gene for non-destructive quantification of promoter activity via color histogram analysis in grapevine (Vitis vinifera) and tobacco.
Identifieur interne : 000546 ( Main/Exploration ); précédent : 000545; suivant : 000547Use of the VvMybA1 gene for non-destructive quantification of promoter activity via color histogram analysis in grapevine (Vitis vinifera) and tobacco.
Auteurs : Zhijian T. Li [États-Unis] ; Sadanand A. Dhekney ; Dennis J. GraySource :
- Transgenic research [ 1573-9368 ] ; 2011.
Descripteurs français
- KwdFr :
- Agrobacterium (génétique), Anthocyanes (biosynthèse), Anthocyanes (génétique), Facteurs de transcription (génétique), Fluorimétrie (MeSH), Gènes rapporteurs (MeSH), Régions promotrices (génétique) (MeSH), Régulation de l'expression des gènes végétaux (MeSH), Tabac (génétique), Vecteurs génétiques (MeSH), Vitis (génétique), Végétaux génétiquement modifiés (génétique).
- MESH :
English descriptors
- KwdEn :
- Agrobacterium (genetics), Anthocyanins (biosynthesis), Anthocyanins (genetics), Fluorometry (MeSH), Gene Expression Regulation, Plant (MeSH), Genes, Reporter (MeSH), Genetic Vectors (MeSH), Plants, Genetically Modified (genetics), Promoter Regions, Genetic (MeSH), Tobacco (genetics), Transcription Factors (genetics), Vitis (genetics).
- MESH :
- chemical , biosynthesis : Anthocyanins.
- genetics : Agrobacterium, Anthocyanins, Plants, Genetically Modified, Tobacco, Transcription Factors, Vitis.
- Fluorometry, Gene Expression Regulation, Plant, Genes, Reporter, Genetic Vectors, Promoter Regions, Genetic.
Abstract
We report the development of a convenient plant-based reporter system to analyze promoters and facilitate selection of genetically engineered plants. The VvMybA1 gene of grapevine (Vitis vinifera L.) regulates the last metabolic step of anthocyanin biosynthesis and its ectopic expression leads to anthocyanin production in otherwise non-pigmented cells. To develop an anthocyanin-based quantitative reporter system, the VvMybA1 gene was isolated from V. vinifera 'Merlot' and placed under control of three promoters to test its ability to distinguish different activity levels. Promoters included a double enhanced CaMV35S (d35S) promoter, a double enhanced CsVMV (dCsVMV) promoter or a bi-directional dual promoter (BDDP), resulting in transformation vectors DAT, CAT and DEAT, respectively. These vectors were introduced into grapevine and tobacco via Agrobacterium-mediated transformation for transient and stable expression analysis. A linear relationship between the mean red brightness (MRB) and optical density (OD) values with a 0.99 regression coefficient was identified in a dilution series of anthocyanin, thus allowing the use of histogram data for non-destructive and real-time assessment of transcriptional activity. Results of histogram-based analysis of color images from transformed grapevine somatic embryos (SE) and various tissues of transgenic tobacco showed a consistent six to sevenfold promoter activity increase of DEAT over DAT. This expression increase was verified by spectroscopic measurement of anthocyanin concentrations in sepal tissue of transgenic tobacco plants. These results were congruent with previously findings of promoter activity derived from GUS fluorometric assay, thus demonstrating for the first time that the VvMybA1 gene could offer a simple, versatile and reliable plant-based alternative for quantitative promoter analysis in plants.
DOI: 10.1007/s11248-010-9482-6
PubMed: 21229312
Affiliations:
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Li</name><affiliation wicri:level="2"><nlm:affiliation>Grape Biotechnology Core Laboratory, Mid-Florida Research and Education Center, University of Florida/IFAS, 2725 Binion Road, Apopka, FL 32703-8504, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Grape Biotechnology Core Laboratory, Mid-Florida Research and Education Center, University of Florida/IFAS, 2725 Binion Road, Apopka, FL 32703-8504</wicri:regionArea><placeName><region type="state">Floride</region></placeName></affiliation></author><author><name sortKey="Dhekney, Sadanand A" sort="Dhekney, Sadanand A" uniqKey="Dhekney S" first="Sadanand A" last="Dhekney">Sadanand A. Dhekney</name></author><author><name sortKey="Gray, Dennis J" sort="Gray, Dennis J" uniqKey="Gray D" first="Dennis J" last="Gray">Dennis J. Gray</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2011">2011</date><idno type="RBID">pubmed:21229312</idno><idno type="pmid">21229312</idno><idno type="doi">10.1007/s11248-010-9482-6</idno><idno type="wicri:Area/Main/Corpus">000576</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">000576</idno><idno type="wicri:Area/Main/Curation">000576</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Curation">000576</idno><idno type="wicri:Area/Main/Exploration">000576</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Use of the VvMybA1 gene for non-destructive quantification of promoter activity via color histogram analysis in grapevine (Vitis vinifera) and tobacco.</title><author><name sortKey="Li, Zhijian T" sort="Li, Zhijian T" uniqKey="Li Z" first="Zhijian T" last="Li">Zhijian T. Li</name><affiliation wicri:level="2"><nlm:affiliation>Grape Biotechnology Core Laboratory, Mid-Florida Research and Education Center, University of Florida/IFAS, 2725 Binion Road, Apopka, FL 32703-8504, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Grape Biotechnology Core Laboratory, Mid-Florida Research and Education Center, University of Florida/IFAS, 2725 Binion Road, Apopka, FL 32703-8504</wicri:regionArea><placeName><region type="state">Floride</region></placeName></affiliation></author><author><name sortKey="Dhekney, Sadanand A" sort="Dhekney, Sadanand A" uniqKey="Dhekney S" first="Sadanand A" last="Dhekney">Sadanand A. Dhekney</name></author><author><name sortKey="Gray, Dennis J" sort="Gray, Dennis J" uniqKey="Gray D" first="Dennis J" last="Gray">Dennis J. Gray</name></author></analytic><series><title level="j">Transgenic research</title><idno type="eISSN">1573-9368</idno><imprint><date when="2011" type="published">2011</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Agrobacterium (genetics)</term><term>Anthocyanins (biosynthesis)</term><term>Anthocyanins (genetics)</term><term>Fluorometry (MeSH)</term><term>Gene Expression Regulation, Plant (MeSH)</term><term>Genes, Reporter (MeSH)</term><term>Genetic Vectors (MeSH)</term><term>Plants, Genetically Modified (genetics)</term><term>Promoter Regions, Genetic (MeSH)</term><term>Tobacco (genetics)</term><term>Transcription Factors (genetics)</term><term>Vitis (genetics)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Agrobacterium (génétique)</term><term>Anthocyanes (biosynthèse)</term><term>Anthocyanes (génétique)</term><term>Facteurs de transcription (génétique)</term><term>Fluorimétrie (MeSH)</term><term>Gènes rapporteurs (MeSH)</term><term>Régions promotrices (génétique) (MeSH)</term><term>Régulation de l'expression des gènes végétaux (MeSH)</term><term>Tabac (génétique)</term><term>Vecteurs génétiques (MeSH)</term><term>Vitis (génétique)</term><term>Végétaux génétiquement modifiés (génétique)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Anthocyanins</term></keywords><keywords scheme="MESH" qualifier="biosynthèse" xml:lang="fr"><term>Anthocyanes</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Agrobacterium</term><term>Anthocyanins</term><term>Plants, Genetically Modified</term><term>Tobacco</term><term>Transcription Factors</term><term>Vitis</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Agrobacterium</term><term>Anthocyanes</term><term>Facteurs de transcription</term><term>Tabac</term><term>Vitis</term><term>Végétaux génétiquement modifiés</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Fluorometry</term><term>Gene Expression Regulation, Plant</term><term>Genes, Reporter</term><term>Genetic Vectors</term><term>Promoter Regions, Genetic</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Fluorimétrie</term><term>Gènes rapporteurs</term><term>Régions promotrices (génétique)</term><term>Régulation de l'expression des gènes végétaux</term><term>Vecteurs génétiques</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">We report the development of a convenient plant-based reporter system to analyze promoters and facilitate selection of genetically engineered plants. The VvMybA1 gene of grapevine (Vitis vinifera L.) regulates the last metabolic step of anthocyanin biosynthesis and its ectopic expression leads to anthocyanin production in otherwise non-pigmented cells. To develop an anthocyanin-based quantitative reporter system, the VvMybA1 gene was isolated from V. vinifera 'Merlot' and placed under control of three promoters to test its ability to distinguish different activity levels. Promoters included a double enhanced CaMV35S (d35S) promoter, a double enhanced CsVMV (dCsVMV) promoter or a bi-directional dual promoter (BDDP), resulting in transformation vectors DAT, CAT and DEAT, respectively. These vectors were introduced into grapevine and tobacco via Agrobacterium-mediated transformation for transient and stable expression analysis. A linear relationship between the mean red brightness (MRB) and optical density (OD) values with a 0.99 regression coefficient was identified in a dilution series of anthocyanin, thus allowing the use of histogram data for non-destructive and real-time assessment of transcriptional activity. Results of histogram-based analysis of color images from transformed grapevine somatic embryos (SE) and various tissues of transgenic tobacco showed a consistent six to sevenfold promoter activity increase of DEAT over DAT. This expression increase was verified by spectroscopic measurement of anthocyanin concentrations in sepal tissue of transgenic tobacco plants. These results were congruent with previously findings of promoter activity derived from GUS fluorometric assay, thus demonstrating for the first time that the VvMybA1 gene could offer a simple, versatile and reliable plant-based alternative for quantitative promoter analysis in plants.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">21229312</PMID><DateCompleted><Year>2012</Year><Month>01</Month><Day>13</Day></DateCompleted><DateRevised><Year>2018</Year><Month>11</Month><Day>13</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1573-9368</ISSN><JournalIssue CitedMedium="Internet"><Volume>20</Volume><Issue>5</Issue><PubDate><Year>2011</Year><Month>Oct</Month></PubDate></JournalIssue><Title>Transgenic research</Title><ISOAbbreviation>Transgenic Res</ISOAbbreviation></Journal><ArticleTitle>Use of the VvMybA1 gene for non-destructive quantification of promoter activity via color histogram analysis in grapevine (Vitis vinifera) and tobacco.</ArticleTitle><Pagination><MedlinePgn>1087-97</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1007/s11248-010-9482-6</ELocationID><Abstract><AbstractText>We report the development of a convenient plant-based reporter system to analyze promoters and facilitate selection of genetically engineered plants. The VvMybA1 gene of grapevine (Vitis vinifera L.) regulates the last metabolic step of anthocyanin biosynthesis and its ectopic expression leads to anthocyanin production in otherwise non-pigmented cells. To develop an anthocyanin-based quantitative reporter system, the VvMybA1 gene was isolated from V. vinifera 'Merlot' and placed under control of three promoters to test its ability to distinguish different activity levels. Promoters included a double enhanced CaMV35S (d35S) promoter, a double enhanced CsVMV (dCsVMV) promoter or a bi-directional dual promoter (BDDP), resulting in transformation vectors DAT, CAT and DEAT, respectively. These vectors were introduced into grapevine and tobacco via Agrobacterium-mediated transformation for transient and stable expression analysis. A linear relationship between the mean red brightness (MRB) and optical density (OD) values with a 0.99 regression coefficient was identified in a dilution series of anthocyanin, thus allowing the use of histogram data for non-destructive and real-time assessment of transcriptional activity. Results of histogram-based analysis of color images from transformed grapevine somatic embryos (SE) and various tissues of transgenic tobacco showed a consistent six to sevenfold promoter activity increase of DEAT over DAT. This expression increase was verified by spectroscopic measurement of anthocyanin concentrations in sepal tissue of transgenic tobacco plants. These results were congruent with previously findings of promoter activity derived from GUS fluorometric assay, thus demonstrating for the first time that the VvMybA1 gene could offer a simple, versatile and reliable plant-based alternative for quantitative promoter analysis in plants.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Li</LastName><ForeName>Zhijian T</ForeName><Initials>ZT</Initials><AffiliationInfo><Affiliation>Grape Biotechnology Core Laboratory, Mid-Florida Research and Education Center, University of Florida/IFAS, 2725 Binion Road, Apopka, FL 32703-8504, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Dhekney</LastName><ForeName>Sadanand A</ForeName><Initials>SA</Initials></Author><Author ValidYN="Y"><LastName>Gray</LastName><ForeName>Dennis J</ForeName><Initials>DJ</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal 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Dhekney</name><name sortKey="Gray, Dennis J" sort="Gray, Dennis J" uniqKey="Gray D" first="Dennis J" last="Gray">Dennis J. Gray</name></noCountry><country name="États-Unis"><region name="Floride"><name sortKey="Li, Zhijian T" sort="Li, Zhijian T" uniqKey="Li Z" first="Zhijian T" last="Li">Zhijian T. Li</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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