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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Tyloses and Phenolic Deposits in Xylem Vessels Impede Water Transport in Low-Lignin Transgenic Poplars: A Study by Cryo-Fluorescence Microscopy<xref ref-type="author-notes" rid="fn1"><sup>1</sup></xref><xref ref-type="author-notes" rid="fn2"><sup>[W]</sup></xref><xref ref-type="author-notes" rid="fn3"><sup>[OA]</sup></xref></title><author><name sortKey="Kitin, Peter" sort="Kitin, Peter" uniqKey="Kitin P" first="Peter" last="Kitin">Peter Kitin</name></author><author><name sortKey="Voelker, Steven L" sort="Voelker, Steven L" uniqKey="Voelker S" first="Steven L." last="Voelker">Steven L. Voelker</name></author><author><name sortKey="Meinzer, Frederick C" sort="Meinzer, Frederick C" uniqKey="Meinzer F" first="Frederick C." last="Meinzer">Frederick C. Meinzer</name></author><author><name sortKey="Beeckman, Hans" sort="Beeckman, Hans" uniqKey="Beeckman H" first="Hans" last="Beeckman">Hans Beeckman</name></author><author><name sortKey="Strauss, Steven H" sort="Strauss, Steven H" uniqKey="Strauss S" first="Steven H." last="Strauss">Steven H. Strauss</name></author><author><name sortKey="Lachenbruch, Barbara" sort="Lachenbruch, Barbara" uniqKey="Lachenbruch B" first="Barbara" last="Lachenbruch">Barbara Lachenbruch</name></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">20639405</idno><idno type="pmc">2949004</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2949004</idno><idno type="RBID">PMC:2949004</idno><idno type="doi">10.1104/pp.110.156224</idno><date when="2010">2010</date><idno type="wicri:Area/Pmc/Corpus">000149</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000149</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Tyloses and Phenolic Deposits in Xylem Vessels Impede Water Transport in Low-Lignin Transgenic Poplars: A Study by Cryo-Fluorescence Microscopy<xref ref-type="author-notes" rid="fn1"><sup>1</sup></xref><xref ref-type="author-notes" rid="fn2"><sup>[W]</sup></xref><xref ref-type="author-notes" rid="fn3"><sup>[OA]</sup></xref></title><author><name sortKey="Kitin, Peter" sort="Kitin, Peter" uniqKey="Kitin P" first="Peter" last="Kitin">Peter Kitin</name></author><author><name sortKey="Voelker, Steven L" sort="Voelker, Steven L" uniqKey="Voelker S" first="Steven L." last="Voelker">Steven L. Voelker</name></author><author><name sortKey="Meinzer, Frederick C" sort="Meinzer, Frederick C" uniqKey="Meinzer F" first="Frederick C." last="Meinzer">Frederick C. Meinzer</name></author><author><name sortKey="Beeckman, Hans" sort="Beeckman, Hans" uniqKey="Beeckman H" first="Hans" last="Beeckman">Hans Beeckman</name></author><author><name sortKey="Strauss, Steven H" sort="Strauss, Steven H" uniqKey="Strauss S" first="Steven H." last="Strauss">Steven H. Strauss</name></author><author><name sortKey="Lachenbruch, Barbara" sort="Lachenbruch, Barbara" uniqKey="Lachenbruch B" first="Barbara" last="Lachenbruch">Barbara Lachenbruch</name></author></analytic><series><title level="j">Plant Physiology</title><idno type="ISSN">0032-0889</idno><idno type="eISSN">1532-2548</idno><imprint><date when="2010">2010</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Of 14 transgenic poplar genotypes (<italic>Populus tremula</italic> × <italic>Populus alba</italic>) with antisense 4-coumarate:coenzyme A ligase that were grown in the field for 2 years, five that had substantial lignin reductions also had greatly reduced xylem-specific conductivity compared with that of control trees and those transgenic events with small reductions in lignin. For the two events with the lowest xylem lignin contents (greater than 40% reduction), we used light microscopy methods and acid fuchsin dye ascent studies to clarify what caused their reduced transport efficiency. A novel protocol involving dye stabilization and cryo-fluorescence microscopy enabled us to visualize the dye at the cellular level and to identify water-conducting pathways in the xylem. Cryo-fixed branch segments were planed in the frozen state on a sliding cryo-microtome and observed with an epifluorescence microscope equipped with a cryo-stage. We could then distinguish clearly between phenolic-occluded vessels, conductive (stain-filled) vessels, and nonconductive (water- or gas-filled) vessels. Low-lignin trees contained areas of nonconductive, brown xylem with patches of collapsed cells and patches of noncollapsed cells filled with phenolics. In contrast, phenolics and nonconductive vessels were rarely observed in normal colored wood of the low-lignin events. The results of cryo-fluorescence light microscopy were supported by observations with a confocal microscope after freeze drying of cryo-planed samples. Moreover, after extraction of the phenolics, confocal microscopy revealed that many of the vessels in the nonconductive xylem were blocked with tyloses. We conclude that reduced transport efficiency of the transgenic low-lignin xylem was largely caused by blockages from tyloses and phenolic deposits within vessels rather than by xylem collapse.</p></div></front></TEI><pmc article-type="research-article"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Plant Physiol</journal-id><journal-id journal-id-type="hwp">plantphysiol</journal-id><journal-id journal-id-type="publisher-id">aspb</journal-id><journal-title-group><journal-title>Plant Physiology</journal-title></journal-title-group><issn pub-type="ppub">0032-0889</issn><issn pub-type="epub">1532-2548</issn><publisher><publisher-name>American Society of Plant Biologists</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">20639405</article-id><article-id pub-id-type="pmc">2949004</article-id><article-id pub-id-type="publisher-id">156224</article-id><article-id pub-id-type="doi">10.1104/pp.110.156224</article-id><article-categories><subj-group><subject>Article</subject></subj-group><subj-group subj-group-type="heading"><subject>Whole Plant and Ecophysiology</subject></subj-group></article-categories><title-group><article-title>Tyloses and Phenolic Deposits in Xylem Vessels Impede Water Transport in Low-Lignin Transgenic Poplars: A Study by Cryo-Fluorescence Microscopy<xref ref-type="author-notes" rid="fn1"><sup>1</sup></xref><xref ref-type="author-notes" rid="fn2"><sup>[W]</sup></xref><xref ref-type="author-notes" rid="fn3"><sup>[OA]</sup></xref></article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Kitin</surname><given-names>Peter</given-names></name><xref ref-type="corresp" rid="cor1"><sup>*</sup></xref></contrib><contrib contrib-type="author"><name><surname>Voelker</surname><given-names>Steven L.</given-names></name></contrib><contrib contrib-type="author"><name><surname>Meinzer</surname><given-names>Frederick C.</given-names></name></contrib><contrib contrib-type="author"><name><surname>Beeckman</surname><given-names>Hans</given-names></name></contrib><contrib contrib-type="author"><name><surname>Strauss</surname><given-names>Steven H.</given-names></name></contrib><contrib contrib-type="author"><name><surname>Lachenbruch</surname><given-names>Barbara</given-names></name></contrib></contrib-group><aff>Laboratory for Wood Biology and Xylarium, Royal Museum for Central Africa, B–3080 Tervuren, Belgium (P.K., H.B.); Department of Wood Science and Engineering (S.L.V., B.L.) and Department of Forest Ecosystems and Society (S.H.S.), Oregon State University, Corvallis, Oregon 97331; United States Department of Agriculture Forest Service, Pacific Northwest Research Station, Corvallis, Oregon 97331 (F.C.M.)</aff><author-notes><corresp id="cor1"><label>*</label>Corresponding author; e-mail <email>peter.kitin@oregonstate.edu</email>.</corresp><fn id="fn1"><label>1</label><p>This work was supported by a Marie Curie Outgoing International Fellowship of the European Commission (no. MOIF 040400 to P.K.) and by an award from the National Institutes of Health to the Confocal Microscopy Facility of the Center for Genome Research and Biocomputing and the Environmental and Health Sciences Center at Oregon State University (grant no. 1S10RR107903–01).</p></fn><fn><p>The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (<ext-link ext-link-type="uri" xlink:href="www.plantphysiol.org">www.plantphysiol.org</ext-link>) is: Peter Kitin (<email>peter.kitin@oregonstate.edu</email>).</p></fn><fn id="fn2"><label>[W]</label><p>The online version of this article contains Web-only data.</p></fn><fn id="fn3"><label>[OA]</label><p>Open Access articles can be viewed online without a subscription.</p></fn><fn><p><ext-link ext-link-type="uri" xlink:href="www.plantphysiol.org/cgi/doi/10.1104/pp.110.156224">www.plantphysiol.org/cgi/doi/10.1104/pp.110.156224</ext-link></p></fn></author-notes><pmc-comment>Fake ppub date generated by PMC from publisher pub-date/@pub-type='epub-ppub' </pmc-comment>
<pub-date pub-type="ppub"><month>10</month><year>2010</year></pub-date><pub-date pub-type="epub"><day>16</day><month>7</month><year>2010</year></pub-date><pub-date pub-type="pmc-release"><day>16</day><month>7</month><year>2010</year></pub-date><pmc-comment> PMC Release delay is 0 months and 0 days and was based on the
<volume>154</volume><issue>2</issue><fpage>887</fpage><lpage>898</lpage><history><date date-type="received"><day>13</day><month>3</month><year>2010</year></date><date date-type="accepted"><day>16</day><month>7</month><year>2010</year></date></history><permissions><copyright-statement>© 2010 American Society of Plant Biologists</copyright-statement><copyright-year>2010</copyright-year></permissions><abstract><p>Of 14 transgenic poplar genotypes (<italic>Populus tremula</italic> × <italic>Populus alba</italic>) with antisense 4-coumarate:coenzyme A ligase that were grown in the field for 2 years, five that had substantial lignin reductions also had greatly reduced xylem-specific conductivity compared with that of control trees and those transgenic events with small reductions in lignin. For the two events with the lowest xylem lignin contents (greater than 40% reduction), we used light microscopy methods and acid fuchsin dye ascent studies to clarify what caused their reduced transport efficiency. A novel protocol involving dye stabilization and cryo-fluorescence microscopy enabled us to visualize the dye at the cellular level and to identify water-conducting pathways in the xylem. Cryo-fixed branch segments were planed in the frozen state on a sliding cryo-microtome and observed with an epifluorescence microscope equipped with a cryo-stage. We could then distinguish clearly between phenolic-occluded vessels, conductive (stain-filled) vessels, and nonconductive (water- or gas-filled) vessels. Low-lignin trees contained areas of nonconductive, brown xylem with patches of collapsed cells and patches of noncollapsed cells filled with phenolics. In contrast, phenolics and nonconductive vessels were rarely observed in normal colored wood of the low-lignin events. The results of cryo-fluorescence light microscopy were supported by observations with a confocal microscope after freeze drying of cryo-planed samples. Moreover, after extraction of the phenolics, confocal microscopy revealed that many of the vessels in the nonconductive xylem were blocked with tyloses. We conclude that reduced transport efficiency of the transgenic low-lignin xylem was largely caused by blockages from tyloses and phenolic deposits within vessels rather than by xylem collapse.</p></abstract></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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