Accurate and high resolution in situ hybridization analysis of gene expression in secondary stem tissues.
Identifieur interne : 004907 ( Main/Exploration ); précédent : 004906; suivant : 004908Accurate and high resolution in situ hybridization analysis of gene expression in secondary stem tissues.
Auteurs : S. Regan ; V. Bourquin ; H. Tuominen ; B. SundbergSource :
- The Plant journal : for cell and molecular biology [ 0960-7412 ] ; 1999.
Descripteurs français
- KwdFr :
- ARN des plantes (génétique), ARN des plantes (métabolisme), ARN messager (génétique), ARN messager (métabolisme), Données de séquences moléculaires (MeSH), Expression des gènes (MeSH), Glucuronidase (génétique), Gènes de plante (MeSH), Hybridation in situ (méthodes), Plantes (génétique), Plantes (métabolisme), Rhizobium (génétique), Tiges de plante (métabolisme), Transformation génétique (MeSH), Végétaux génétiquement modifiés (MeSH), bêta-Glucosidase (génétique).
- MESH :
- génétique : ARN des plantes, ARN messager, Glucuronidase, Plantes, Rhizobium, bêta-Glucosidase.
- métabolisme : ARN des plantes, ARN messager, Plantes, Tiges de plante.
- méthodes : Hybridation in situ.
- Données de séquences moléculaires, Expression des gènes, Gènes de plante, Transformation génétique, Végétaux génétiquement modifiés.
English descriptors
- KwdEn :
- Gene Expression (MeSH), Genes, Plant (MeSH), Glucuronidase (genetics), In Situ Hybridization (methods), Molecular Sequence Data (MeSH), Plant Stems (metabolism), Plants (genetics), Plants (metabolism), Plants, Genetically Modified (MeSH), RNA, Messenger (genetics), RNA, Messenger (metabolism), RNA, Plant (genetics), RNA, Plant (metabolism), Rhizobium (genetics), Transformation, Genetic (MeSH), beta-Glucosidase (genetics).
- MESH :
- chemical , genetics : Glucuronidase, RNA, Messenger, RNA, Plant, beta-Glucosidase.
- genetics : Plants, Rhizobium.
- metabolism : Plant Stems, Plants, RNA, Messenger, RNA, Plant.
- methods : In Situ Hybridization.
- Gene Expression, Genes, Plant, Molecular Sequence Data, Plants, Genetically Modified, Transformation, Genetic.
Abstract
Accurate in situ hybridization analysis in secondary stem tissues of plants has been hindered by specific characteristics of these tissues. First, secondary cell walls non-specifically bind probes used for in situ hybridization thus preventing gene expression analysis in the lignified regions of the stem, such as the xylem. Second, the mRNA in the cambial meristem and its recent derivatives are prone to inadequate fixation when conventional techniques are used. Here we describe an in situ hybridization technique which uses fast freezing and freeze substitution to cryoimmobilize the mRNA followed by embedding in a methacrylate resin for high-resolution analysis of gene expression. By using a transgenic poplar line harbouring rolC:uidA, rolC:iaaM, the gene expression pattern could be compared with histochemical GUS staining. This in situ hybridization technique results in superior preservation of cellular contents, retention of mRNA in all cell types in the poplar stem, a significant reduction of non-specific binding to secondary cell walls and a resolution not previously possible in secondary tissues. This technique will be particularly valuable for the expression analysis of genes involved in xylogenesis and wood formation.
DOI: 10.1046/j.1365-313x.1999.00536.x
PubMed: 10476083
Affiliations:
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Sundberg</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="1999">1999</date><idno type="RBID">pubmed:10476083</idno><idno type="pmid">10476083</idno><idno type="doi">10.1046/j.1365-313x.1999.00536.x</idno><idno type="wicri:Area/Main/Corpus">004904</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">004904</idno><idno type="wicri:Area/Main/Curation">004904</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Curation">004904</idno><idno type="wicri:Area/Main/Exploration">004904</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Accurate and high resolution in situ hybridization analysis of gene expression in secondary stem tissues.</title><author><name sortKey="Regan, S" sort="Regan, S" uniqKey="Regan S" first="S" last="Regan">S. Regan</name><affiliation><nlm:affiliation>Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences, Umeå.</nlm:affiliation><wicri:noCountry code="subField">Umeå</wicri:noCountry></affiliation></author><author><name sortKey="Bourquin, V" sort="Bourquin, V" uniqKey="Bourquin V" first="V" last="Bourquin">V. Bourquin</name></author><author><name sortKey="Tuominen, H" sort="Tuominen, H" uniqKey="Tuominen H" first="H" last="Tuominen">H. Tuominen</name></author><author><name sortKey="Sundberg, B" sort="Sundberg, B" uniqKey="Sundberg B" first="B" last="Sundberg">B. Sundberg</name></author></analytic><series><title level="j">The Plant journal : for cell and molecular biology</title><idno type="ISSN">0960-7412</idno><imprint><date when="1999" type="published">1999</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Gene Expression (MeSH)</term><term>Genes, Plant (MeSH)</term><term>Glucuronidase (genetics)</term><term>In Situ Hybridization (methods)</term><term>Molecular Sequence Data (MeSH)</term><term>Plant Stems (metabolism)</term><term>Plants (genetics)</term><term>Plants (metabolism)</term><term>Plants, Genetically Modified (MeSH)</term><term>RNA, Messenger (genetics)</term><term>RNA, Messenger (metabolism)</term><term>RNA, Plant (genetics)</term><term>RNA, Plant (metabolism)</term><term>Rhizobium (genetics)</term><term>Transformation, Genetic (MeSH)</term><term>beta-Glucosidase (genetics)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ARN des plantes (génétique)</term><term>ARN des plantes (métabolisme)</term><term>ARN messager (génétique)</term><term>ARN messager (métabolisme)</term><term>Données de séquences moléculaires (MeSH)</term><term>Expression des gènes (MeSH)</term><term>Glucuronidase (génétique)</term><term>Gènes de plante (MeSH)</term><term>Hybridation in situ (méthodes)</term><term>Plantes (génétique)</term><term>Plantes (métabolisme)</term><term>Rhizobium (génétique)</term><term>Tiges de plante (métabolisme)</term><term>Transformation génétique (MeSH)</term><term>Végétaux génétiquement modifiés (MeSH)</term><term>bêta-Glucosidase (génétique)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Glucuronidase</term><term>RNA, Messenger</term><term>RNA, Plant</term><term>beta-Glucosidase</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Plants</term><term>Rhizobium</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ARN des plantes</term><term>ARN messager</term><term>Glucuronidase</term><term>Plantes</term><term>Rhizobium</term><term>bêta-Glucosidase</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Plant Stems</term><term>Plants</term><term>RNA, Messenger</term><term>RNA, Plant</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>In Situ Hybridization</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>ARN des plantes</term><term>ARN messager</term><term>Plantes</term><term>Tiges de plante</term></keywords><keywords scheme="MESH" qualifier="méthodes" xml:lang="fr"><term>Hybridation in situ</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Gene Expression</term><term>Genes, Plant</term><term>Molecular Sequence Data</term><term>Plants, Genetically Modified</term><term>Transformation, Genetic</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Données de séquences moléculaires</term><term>Expression des gènes</term><term>Gènes de plante</term><term>Transformation génétique</term><term>Végétaux génétiquement modifiés</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Accurate in situ hybridization analysis in secondary stem tissues of plants has been hindered by specific characteristics of these tissues. First, secondary cell walls non-specifically bind probes used for in situ hybridization thus preventing gene expression analysis in the lignified regions of the stem, such as the xylem. Second, the mRNA in the cambial meristem and its recent derivatives are prone to inadequate fixation when conventional techniques are used. Here we describe an in situ hybridization technique which uses fast freezing and freeze substitution to cryoimmobilize the mRNA followed by embedding in a methacrylate resin for high-resolution analysis of gene expression. By using a transgenic poplar line harbouring rolC:uidA, rolC:iaaM, the gene expression pattern could be compared with histochemical GUS staining. This in situ hybridization technique results in superior preservation of cellular contents, retention of mRNA in all cell types in the poplar stem, a significant reduction of non-specific binding to secondary cell walls and a resolution not previously possible in secondary tissues. This technique will be particularly valuable for the expression analysis of genes involved in xylogenesis and wood formation.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">10476083</PMID><DateCompleted><Year>1999</Year><Month>10</Month><Day>21</Day></DateCompleted><DateRevised><Year>2019</Year><Month>09</Month><Day>05</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0960-7412</ISSN><JournalIssue CitedMedium="Print"><Volume>19</Volume><Issue>3</Issue><PubDate><Year>1999</Year><Month>Aug</Month></PubDate></JournalIssue><Title>The Plant journal : for cell and molecular biology</Title><ISOAbbreviation>Plant J</ISOAbbreviation></Journal><ArticleTitle>Accurate and high resolution in situ hybridization analysis of gene expression in secondary stem tissues.</ArticleTitle><Pagination><MedlinePgn>363-9</MedlinePgn></Pagination><Abstract><AbstractText>Accurate in situ hybridization analysis in secondary stem tissues of plants has been hindered by specific characteristics of these tissues. First, secondary cell walls non-specifically bind probes used for in situ hybridization thus preventing gene expression analysis in the lignified regions of the stem, such as the xylem. Second, the mRNA in the cambial meristem and its recent derivatives are prone to inadequate fixation when conventional techniques are used. Here we describe an in situ hybridization technique which uses fast freezing and freeze substitution to cryoimmobilize the mRNA followed by embedding in a methacrylate resin for high-resolution analysis of gene expression. By using a transgenic poplar line harbouring rolC:uidA, rolC:iaaM, the gene expression pattern could be compared with histochemical GUS staining. This in situ hybridization technique results in superior preservation of cellular contents, retention of mRNA in all cell types in the poplar stem, a significant reduction of non-specific binding to secondary cell walls and a resolution not previously possible in secondary tissues. This technique will be particularly valuable for the expression analysis of genes involved in xylogenesis and wood formation.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Regan</LastName><ForeName>S</ForeName><Initials>S</Initials><AffiliationInfo><Affiliation>Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences, Umeå.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Bourquin</LastName><ForeName>V</ForeName><Initials>V</Initials></Author><Author ValidYN="Y"><LastName>Tuominen</LastName><ForeName>H</ForeName><Initials>H</Initials></Author><Author ValidYN="Y"><LastName>Sundberg</LastName><ForeName>B</ForeName><Initials>B</Initials></Author></AuthorList><Language>eng</Language><DataBankList CompleteYN="Y"><DataBank><DataBankName>GENBANK</DataBankName><AccessionNumberList><AccessionNumber>AI161452</AccessionNumber></AccessionNumberList></DataBank></DataBankList><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>England</Country><MedlineTA>Plant J</MedlineTA><NlmUniqueID>9207397</NlmUniqueID><ISSNLinking>0960-7412</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D012333">RNA, Messenger</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D018749">RNA, Plant</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 3.2.1.-</RegistryNumber><NameOfSubstance UI="C071257">cytokinin-beta-glucosidase</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 3.2.1.21</RegistryNumber><NameOfSubstance UI="D001617">beta-Glucosidase</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 3.2.1.31</RegistryNumber><NameOfSubstance UI="D005966">Glucuronidase</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D015870" MajorTopicYN="N">Gene Expression</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D017343" MajorTopicYN="Y">Genes, Plant</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D005966" MajorTopicYN="N">Glucuronidase</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D017403" MajorTopicYN="N">In Situ Hybridization</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D008969" MajorTopicYN="N">Molecular Sequence Data</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D018547" MajorTopicYN="N">Plant Stems</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010944" MajorTopicYN="N">Plants</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D030821" MajorTopicYN="N">Plants, Genetically Modified</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D012333" MajorTopicYN="N">RNA, Messenger</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D018749" MajorTopicYN="N">RNA, Plant</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D012231" MajorTopicYN="N">Rhizobium</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D014170" MajorTopicYN="N">Transformation, Genetic</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D001617" MajorTopicYN="N">beta-Glucosidase</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>1999</Year><Month>9</Month><Day>4</Day></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>1999</Year><Month>9</Month><Day>4</Day><Hour>0</Hour><Minute>1</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>1999</Year><Month>9</Month><Day>4</Day><Hour>0</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">10476083</ArticleId><ArticleId IdType="pii">tpj536</ArticleId><ArticleId IdType="doi">10.1046/j.1365-313x.1999.00536.x</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list></list><tree><noCountry><name sortKey="Bourquin, V" sort="Bourquin, V" uniqKey="Bourquin V" first="V" last="Bourquin">V. Bourquin</name><name sortKey="Regan, S" sort="Regan, S" uniqKey="Regan S" first="S" last="Regan">S. Regan</name><name sortKey="Sundberg, B" sort="Sundberg, B" uniqKey="Sundberg B" first="B" last="Sundberg">B. Sundberg</name><name sortKey="Tuominen, H" sort="Tuominen, H" uniqKey="Tuominen H" first="H" last="Tuominen">H. Tuominen</name></noCountry></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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