Neutral red as a probe for confocal laser scanning microscopy studies of plant roots.
Identifieur interne : 003331 ( Main/Corpus ); précédent : 003330; suivant : 003332Neutral red as a probe for confocal laser scanning microscopy studies of plant roots.
Auteurs : Joseph G. Dubrovsky ; Martin Guttenberger ; Andres Saralegui ; Selene Napsucialy-Mendivil ; Boris Voigt ; Frantisek Baluska ; Diedrik MenzelSource :
- Annals of botany [ 0305-7364 ] ; 2006.
English descriptors
- KwdEn :
- MESH :
- chemical : Neutral Red.
- anatomy & histology : Plant Roots.
- cytology : Mycorrhizae.
- growth & development : Plant Roots.
- methods : Microscopy, Confocal.
- ultrastructure : Vacuoles.
- Molecular Probe Techniques.
Abstract
BACKGROUND AND AIMS
Neutral red (NR), a lipophilic phenazine dye, has been widely used in various biological systems as a vital stain for bright-field microscopy. In its unprotonated form it penetrates the plasma membrane and tonoplast of viable plant cells, then due to protonation it becomes trapped in acidic compartments. The possible applications of NR for confocal laser scanning microscopy (CLSM) studies were examined in various aspects of plant root biology.
METHODS
NR was used as a fluorochrome for living roots of Phaseolus vulgaris, Allium cepa, A. porrum and Arabidopsis thaliana (wild-type and transgenic GFP-carrying lines). The tissues were visualized using CLSM. The effect of NR on the integrity of the cytoskeleton and the growth rate of arabidopsis primary roots was analysed to judge potential toxic effects of the dye.
KEY RESULTS
The main advantages of the use of NR are related to the fact that NR rapidly penetrates root tissues, has affinity to suberin and lignin, and accumulates in the vacuoles. It is shown that NR is a suitable probe for visualization of proto- and metaxylem elements, Casparian bands in the endodermis, and vacuoles in cells of living roots. The actin cytoskeleton and the microtubule system of the cells, as well as the dynamics of root growth, remain unchanged after short-term application of NR, indicating a relatively low toxicity of this chemical. It was also found that NR is a useful probe for the observation of the internal structures of root nodules and of fungal hyphae in vesicular-arbuscular mycorrhizas.
CONCLUSIONS
Ease, low cost and absence of tissue processing make NR a useful probe for structural, developmental and vacuole-biogenetic studies of plant roots with CLSM.
DOI: 10.1093/aob/mcl045
PubMed: 16520341
PubMed Central: PMC2803381
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Dubrovsky</name><affiliation><nlm:affiliation>Departamento de Biología Molecular de Plantas, Universidad Nacional Autónoma de México, Cuernavaca, Morelos, México. jdubrov@ibt.unam.mx</nlm:affiliation></affiliation></author><author><name sortKey="Guttenberger, Martin" sort="Guttenberger, Martin" uniqKey="Guttenberger M" first="Martin" last="Guttenberger">Martin Guttenberger</name></author><author><name sortKey="Saralegui, Andres" sort="Saralegui, Andres" uniqKey="Saralegui A" first="Andres" last="Saralegui">Andres Saralegui</name></author><author><name sortKey="Napsucialy Mendivil, Selene" sort="Napsucialy Mendivil, Selene" uniqKey="Napsucialy Mendivil S" first="Selene" last="Napsucialy-Mendivil">Selene Napsucialy-Mendivil</name></author><author><name sortKey="Voigt, Boris" sort="Voigt, Boris" uniqKey="Voigt B" first="Boris" last="Voigt">Boris Voigt</name></author><author><name sortKey="Baluska, Frantisek" sort="Baluska, Frantisek" uniqKey="Baluska F" first="Frantisek" last="Baluska">Frantisek Baluska</name></author><author><name sortKey="Menzel, Diedrik" sort="Menzel, Diedrik" uniqKey="Menzel D" first="Diedrik" last="Menzel">Diedrik Menzel</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2006">2006</date><idno type="RBID">pubmed:16520341</idno><idno type="pmid">16520341</idno><idno type="doi">10.1093/aob/mcl045</idno><idno type="pmc">PMC2803381</idno><idno type="wicri:Area/Main/Corpus">003331</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">003331</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Neutral red as a probe for confocal laser scanning microscopy studies of plant roots.</title><author><name sortKey="Dubrovsky, Joseph G" sort="Dubrovsky, Joseph G" uniqKey="Dubrovsky J" first="Joseph G" last="Dubrovsky">Joseph G. Dubrovsky</name><affiliation><nlm:affiliation>Departamento de Biología Molecular de Plantas, Universidad Nacional Autónoma de México, Cuernavaca, Morelos, México. jdubrov@ibt.unam.mx</nlm:affiliation></affiliation></author><author><name sortKey="Guttenberger, Martin" sort="Guttenberger, Martin" uniqKey="Guttenberger M" first="Martin" last="Guttenberger">Martin Guttenberger</name></author><author><name sortKey="Saralegui, Andres" sort="Saralegui, Andres" uniqKey="Saralegui A" first="Andres" last="Saralegui">Andres Saralegui</name></author><author><name sortKey="Napsucialy Mendivil, Selene" sort="Napsucialy Mendivil, Selene" uniqKey="Napsucialy Mendivil S" first="Selene" last="Napsucialy-Mendivil">Selene Napsucialy-Mendivil</name></author><author><name sortKey="Voigt, Boris" sort="Voigt, Boris" uniqKey="Voigt B" first="Boris" last="Voigt">Boris Voigt</name></author><author><name sortKey="Baluska, Frantisek" sort="Baluska, Frantisek" uniqKey="Baluska F" first="Frantisek" last="Baluska">Frantisek Baluska</name></author><author><name sortKey="Menzel, Diedrik" sort="Menzel, Diedrik" uniqKey="Menzel D" first="Diedrik" last="Menzel">Diedrik Menzel</name></author></analytic><series><title level="j">Annals of botany</title><idno type="ISSN">0305-7364</idno><imprint><date when="2006" type="published">2006</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Microscopy, Confocal (methods)</term><term>Molecular Probe Techniques (MeSH)</term><term>Mycorrhizae (cytology)</term><term>Neutral Red (MeSH)</term><term>Plant Roots (anatomy & histology)</term><term>Plant Roots (growth & development)</term><term>Vacuoles (ultrastructure)</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>Neutral Red</term></keywords><keywords scheme="MESH" qualifier="anatomy & histology" xml:lang="en"><term>Plant Roots</term></keywords><keywords scheme="MESH" qualifier="cytology" xml:lang="en"><term>Mycorrhizae</term></keywords><keywords scheme="MESH" qualifier="growth & development" xml:lang="en"><term>Plant Roots</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Microscopy, Confocal</term></keywords><keywords scheme="MESH" qualifier="ultrastructure" xml:lang="en"><term>Vacuoles</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Molecular Probe Techniques</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p><b>BACKGROUND AND AIMS</b></p><p>Neutral red (NR), a lipophilic phenazine dye, has been widely used in various biological systems as a vital stain for bright-field microscopy. In its unprotonated form it penetrates the plasma membrane and tonoplast of viable plant cells, then due to protonation it becomes trapped in acidic compartments. The possible applications of NR for confocal laser scanning microscopy (CLSM) studies were examined in various aspects of plant root biology.</p></div><div type="abstract" xml:lang="en"><p><b>METHODS</b></p><p>NR was used as a fluorochrome for living roots of Phaseolus vulgaris, Allium cepa, A. porrum and Arabidopsis thaliana (wild-type and transgenic GFP-carrying lines). The tissues were visualized using CLSM. The effect of NR on the integrity of the cytoskeleton and the growth rate of arabidopsis primary roots was analysed to judge potential toxic effects of the dye.</p></div><div type="abstract" xml:lang="en"><p><b>KEY RESULTS</b></p><p>The main advantages of the use of NR are related to the fact that NR rapidly penetrates root tissues, has affinity to suberin and lignin, and accumulates in the vacuoles. It is shown that NR is a suitable probe for visualization of proto- and metaxylem elements, Casparian bands in the endodermis, and vacuoles in cells of living roots. The actin cytoskeleton and the microtubule system of the cells, as well as the dynamics of root growth, remain unchanged after short-term application of NR, indicating a relatively low toxicity of this chemical. It was also found that NR is a useful probe for the observation of the internal structures of root nodules and of fungal hyphae in vesicular-arbuscular mycorrhizas.</p></div><div type="abstract" xml:lang="en"><p><b>CONCLUSIONS</b></p><p>Ease, low cost and absence of tissue processing make NR a useful probe for structural, developmental and vacuole-biogenetic studies of plant roots with CLSM.</p></div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">16520341</PMID><DateCompleted><Year>2006</Year><Month>09</Month><Day>22</Day></DateCompleted><DateRevised><Year>2018</Year><Month>11</Month><Day>13</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Print">0305-7364</ISSN><JournalIssue CitedMedium="Print"><Volume>97</Volume><Issue>6</Issue><PubDate><Year>2006</Year><Month>Jun</Month></PubDate></JournalIssue><Title>Annals of botany</Title><ISOAbbreviation>Ann Bot</ISOAbbreviation></Journal><ArticleTitle>Neutral red as a probe for confocal laser scanning microscopy studies of plant roots.</ArticleTitle><Pagination><MedlinePgn>1127-38</MedlinePgn></Pagination><Abstract><AbstractText Label="BACKGROUND AND AIMS" NlmCategory="OBJECTIVE">Neutral red (NR), a lipophilic phenazine dye, has been widely used in various biological systems as a vital stain for bright-field microscopy. In its unprotonated form it penetrates the plasma membrane and tonoplast of viable plant cells, then due to protonation it becomes trapped in acidic compartments. The possible applications of NR for confocal laser scanning microscopy (CLSM) studies were examined in various aspects of plant root biology.</AbstractText><AbstractText Label="METHODS" NlmCategory="METHODS">NR was used as a fluorochrome for living roots of Phaseolus vulgaris, Allium cepa, A. porrum and Arabidopsis thaliana (wild-type and transgenic GFP-carrying lines). The tissues were visualized using CLSM. The effect of NR on the integrity of the cytoskeleton and the growth rate of arabidopsis primary roots was analysed to judge potential toxic effects of the dye.</AbstractText><AbstractText Label="KEY RESULTS" NlmCategory="RESULTS">The main advantages of the use of NR are related to the fact that NR rapidly penetrates root tissues, has affinity to suberin and lignin, and accumulates in the vacuoles. It is shown that NR is a suitable probe for visualization of proto- and metaxylem elements, Casparian bands in the endodermis, and vacuoles in cells of living roots. The actin cytoskeleton and the microtubule system of the cells, as well as the dynamics of root growth, remain unchanged after short-term application of NR, indicating a relatively low toxicity of this chemical. It was also found that NR is a useful probe for the observation of the internal structures of root nodules and of fungal hyphae in vesicular-arbuscular mycorrhizas.</AbstractText><AbstractText Label="CONCLUSIONS" NlmCategory="CONCLUSIONS">Ease, low cost and absence of tissue processing make NR a useful probe for structural, developmental and vacuole-biogenetic studies of plant roots with CLSM.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Dubrovsky</LastName><ForeName>Joseph G</ForeName><Initials>JG</Initials><AffiliationInfo><Affiliation>Departamento de Biología Molecular de Plantas, Universidad Nacional Autónoma de México, Cuernavaca, Morelos, México. jdubrov@ibt.unam.mx</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Guttenberger</LastName><ForeName>Martin</ForeName><Initials>M</Initials></Author><Author ValidYN="Y"><LastName>Saralegui</LastName><ForeName>Andres</ForeName><Initials>A</Initials></Author><Author ValidYN="Y"><LastName>Napsucialy-Mendivil</LastName><ForeName>Selene</ForeName><Initials>S</Initials></Author><Author ValidYN="Y"><LastName>Voigt</LastName><ForeName>Boris</ForeName><Initials>B</Initials></Author><Author ValidYN="Y"><LastName>Baluska</LastName><ForeName>Frantisek</ForeName><Initials>F</Initials></Author><Author ValidYN="Y"><LastName>Menzel</LastName><ForeName>Diedrik</ForeName><Initials>D</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>2006</Year><Month>03</Month><Day>06</Day></ArticleDate></Article><MedlineJournalInfo><Country>England</Country><MedlineTA>Ann 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