{"id":{"repo_id":"vt","oai_identifier":"oai:vtechworks.lib.vt.edu:10919/38851"},"canonical_url":"https://search.dev.ndltd.org/etd/vt/oai:vtechworks.lib.vt.edu:10919/38851","repository":{"repo_id":"vt","name":"Virginia Tech","base_url":"https://vtechworks.lib.vt.edu/oai/request"},"display":{"title":"Characterization of lignin deposition in <i>Pinus taeda</i> L. cell suspension cultures","abstract":"<i>Pinus taeda</i> L. suspension culture cells were used to develop a model system to study the process of lignification occurring during the early stages of cell wall formation and maturation. Chemical, biochemical and histochemical analyses of the <i>P. taeda</i> suspension cultures grown with 2,4-dichlorophenoxyacetic acid (2,4-D) as the growth regulator did not provide conclusive evidence for lignin deposition. On the other hand, cultures in which 2,4-D was substituted with α-naphthaleneacetic acid (NAA) were shown to lignify. During this induction of lignification, limited cell wall thickening occurred since transmission electron microscopy of the 2,4-D grown cells showed only primary walls while the average cell wall thickness of the NAA-grown cells was consistent with secondary (S₁) layer formation. Despite the possibility of only limited lignin deposition in the 2,4-0 grown cells, secondary metabolism had occurred as evidenced by reversed-phase and chiral chromatographic separations which revealed the ability of these cells to produce enantiomerically pure (-)-matairesinol. Administrations of [1-¹³C], [2-¹³C ] and [3-¹³C ] specifically labeled phenylalanines to the <i>P. taeda</i> suspension cultures in medium containing NAA allowed the determination of lignin bonding patterns <i>in situ</i> by solid-state ¹³C NMR spectroscopy of the resulting ¹³C enriched cells. Aqueous and organic solvent extractions and protease treatment yielded ¹³C enriched cell walls for solid-state ¹³C NMR spectroscopic analyses of the cell wall bound lignin component. Subsequently, an isolated lignin derivative from these cell walls was analyzed by solution-state ¹³C NMR spectroscopy and verified the assignments made in the solid-state. Accordingly, the above experiments represent the first demonstration of lignin bonding patterns <i>in situ</i> in a <i>Pinus</i> species as well as a suspension culture. This culture system possesses great potential as a model to thoroughly study the early stages of lignification.","abstract_html":"&lt;i&gt;Pinus taeda&lt;/i&gt; L. suspension culture cells were used to develop a model system to study the process of lignification occurring during the early stages of cell wall formation and maturation. Chemical, biochemical and histochemical analyses of the &lt;i&gt;P. taeda&lt;/i&gt; suspension cultures grown with 2,4-dichlorophenoxyacetic acid (2,4-D) as the growth regulator did not provide conclusive evidence for lignin deposition. On the other hand, cultures in which 2,4-D was substituted with α-naphthaleneacetic acid (NAA) were shown to lignify. During this induction of lignification, limited cell wall thickening occurred since transmission electron microscopy of the 2,4-D grown cells showed only primary walls while the average cell wall thickness of the NAA-grown cells was consistent with secondary (S₁) layer formation. Despite the possibility of only limited lignin deposition in the 2,4-0 grown cells, secondary metabolism had occurred as evidenced by reversed-phase and chiral chromatographic separations which revealed the ability of these cells to produce enantiomerically pure (-)-matairesinol. Administrations of [1-¹³C], [2-¹³C ] and [3-¹³C ] specifically labeled phenylalanines to the &lt;i&gt;P. taeda&lt;/i&gt; suspension cultures in medium containing NAA allowed the determination of lignin bonding patterns &lt;i&gt;in situ&lt;/i&gt; by solid-state ¹³C NMR spectroscopy of the resulting ¹³C enriched cells. Aqueous and organic solvent extractions and protease treatment yielded ¹³C enriched cell walls for solid-state ¹³C NMR spectroscopic analyses of the cell wall bound lignin component. Subsequently, an isolated lignin derivative from these cell walls was analyzed by solution-state ¹³C NMR spectroscopy and verified the assignments made in the solid-state. Accordingly, the above experiments represent the first demonstration of lignin bonding patterns &lt;i&gt;in situ&lt;/i&gt; in a &lt;i&gt;Pinus&lt;/i&gt; species as well as a suspension culture. This culture system possesses great potential as a model to thoroughly study the early stages of lignification.","abstract_has_math":false,"creators":["Eberhardt, Thomas Leonard"],"institution":"Virginia Tech","degree_name":"Ph. D.","degree_level":"doctoral","degree_discipline":"Wood Science and Forest Products","degree_department":"Wood Science and Forest Products","school":null,"contributors":[],"advisors":[],"committee_chairs":["Lewis, Norman G."],"committee_members":["Ifju, Geza","Cramer, Carole L.","Glasser, Wolfgang G.","Hess, John L.","White, Robert H."],"year":1992,"date_issued":"1992-03-05","date_published":"1992-03-05","updated_at":"2026-07-22T22:18:55Z","subjects":[],"languages":["en"],"rights":["In Copyright"],"rights_urls":["http://rightsstatements.org/vocab/InC/1.0/"],"identifier_entries":[{"key":"dc:identifier.other","label":"Dc Identifier Other","values":["etd-07282008-134210"],"render_values":[{"text":"etd-07282008-134210","href":null,"code":true}]}]},"links":{"outbound_url":"http://hdl.handle.net/10919/38851","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.committeechair","label":"Committee Chair","values":["Lewis, Norman G."]},{"key":"dc:contributor.committeemember","label":"Committee Member","values":["Ifju, Geza","Cramer, Carole L.","Glasser, Wolfgang G.","Hess, John L.","White, Robert H."]},{"key":"dc:contributor.department","label":"Department","values":["Wood Science and Forest Products"]},{"key":"dc:creator","label":"Author","values":["Eberhardt, Thomas Leonard"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2014-03-14T21:16:20Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2014-03-14T21:16:20Z","2008-07-28"]},{"key":"dc:date.issued","label":"Date","values":["1992-03-05"]},{"key":"dc:publisher","label":"Institution","values":["Virginia Tech"]},{"key":"dc:type","label":"Dc Type","values":["Dissertation"]},{"key":"dc:type.dcmitype","label":"Dc Type Dcmitype","values":["Text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Wood Science and Forest Products"]},{"key":"thesis:degree_level","label":"Degree Level","values":["doctoral"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph. 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Chemical, biochemical and histochemical analyses of the <i>P. taeda</i> suspension cultures grown with 2,4-dichlorophenoxyacetic acid (2,4-D) as the growth regulator did not provide conclusive evidence for lignin deposition. On the other hand, cultures in which 2,4-D was substituted with α-naphthaleneacetic acid (NAA) were shown to lignify. During this induction of lignification, limited cell wall thickening occurred since transmission electron microscopy of the 2,4-D grown cells showed only primary walls while the average cell wall thickness of the NAA-grown cells was consistent with secondary (S₁) layer formation. Despite the possibility of only limited lignin deposition in the 2,4-0 grown cells, secondary metabolism had occurred as evidenced by reversed-phase and chiral chromatographic separations which revealed the ability of these cells to produce enantiomerically pure (-)-matairesinol. Administrations of [1-¹³C], [2-¹³C ] and [3-¹³C ] specifically labeled phenylalanines to the <i>P. taeda</i> suspension cultures in medium containing NAA allowed the determination of lignin bonding patterns <i>in situ</i> by solid-state ¹³C NMR spectroscopy of the resulting ¹³C enriched cells. Aqueous and organic solvent extractions and protease treatment yielded ¹³C enriched cell walls for solid-state ¹³C NMR spectroscopic analyses of the cell wall bound lignin component. Subsequently, an isolated lignin derivative from these cell walls was analyzed by solution-state ¹³C NMR spectroscopy and verified the assignments made in the solid-state. Accordingly, the above experiments represent the first demonstration of lignin bonding patterns <i>in situ</i> in a <i>Pinus</i> species as well as a suspension culture. This culture system possesses great potential as a model to thoroughly study the early stages of lignification."]},{"key":"dc:description.degree","label":"Dc Description Degree","values":["Ph. D."]},{"key":"dc:format.medium","label":"Dc Format Medium","values":["BTD"]},{"key":"dc:format.mimetype","label":"Dc Format Mimetype","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Characterization of lignin deposition in <i>Pinus taeda</i> L. cell suspension cultures"]}]}],"canonical_facts":{"dc:contributor.committeechair":["Lewis, Norman G."],"dc:contributor.committeemember":["Ifju, Geza","Cramer, Carole L.","Glasser, Wolfgang G.","Hess, John L.","White, Robert H."],"dc:contributor.department":["Wood Science and Forest Products"],"dc:creator":["Eberhardt, Thomas Leonard"],"dc:date.accessioned":["2014-03-14T21:16:20Z"],"dc:date.available":["2014-03-14T21:16:20Z","2008-07-28"],"dc:date.issued":["1992-03-05"],"dc:description.abstract":["<i>Pinus taeda</i> L. suspension culture cells were used to develop a model system to study the process of lignification occurring during the early stages of cell wall formation and maturation. Chemical, biochemical and histochemical analyses of the <i>P. taeda</i> suspension cultures grown with 2,4-dichlorophenoxyacetic acid (2,4-D) as the growth regulator did not provide conclusive evidence for lignin deposition. On the other hand, cultures in which 2,4-D was substituted with α-naphthaleneacetic acid (NAA) were shown to lignify. During this induction of lignification, limited cell wall thickening occurred since transmission electron microscopy of the 2,4-D grown cells showed only primary walls while the average cell wall thickness of the NAA-grown cells was consistent with secondary (S₁) layer formation. Despite the possibility of only limited lignin deposition in the 2,4-0 grown cells, secondary metabolism had occurred as evidenced by reversed-phase and chiral chromatographic separations which revealed the ability of these cells to produce enantiomerically pure (-)-matairesinol. Administrations of [1-¹³C], [2-¹³C ] and [3-¹³C ] specifically labeled phenylalanines to the <i>P. taeda</i> suspension cultures in medium containing NAA allowed the determination of lignin bonding patterns <i>in situ</i> by solid-state ¹³C NMR spectroscopy of the resulting ¹³C enriched cells. Aqueous and organic solvent extractions and protease treatment yielded ¹³C enriched cell walls for solid-state ¹³C NMR spectroscopic analyses of the cell wall bound lignin component. Subsequently, an isolated lignin derivative from these cell walls was analyzed by solution-state ¹³C NMR spectroscopy and verified the assignments made in the solid-state. Accordingly, the above experiments represent the first demonstration of lignin bonding patterns <i>in situ</i> in a <i>Pinus</i> species as well as a suspension culture. This culture system possesses great potential as a model to thoroughly study the early stages of lignification."],"dc:description.degree":["Ph. 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