Characterization of lignin deposition in Pinus taeda L. cell suspension cultures

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dc.contributor.authorEberhardt, Thomas Leonarden
dc.contributor.committeechairLewis, Norman G.en
dc.contributor.committeememberIfju, Gezaen
dc.contributor.committeememberCramer, Carole L.en
dc.contributor.committeememberGlasser, Wolfgang G.en
dc.contributor.committeememberHess, John L.en
dc.contributor.committeememberWhite, Robert H.en
dc.contributor.departmentWood Science and Forest Productsen
dc.date.accessioned2014-03-14T21:16:20Zen
dc.date.adate2008-07-28en
dc.date.available2014-03-14T21:16:20Zen
dc.date.issued1992-03-05en
dc.date.rdate2008-07-28en
dc.date.sdate2008-07-28en
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.en
dc.description.degreePh. D.en
dc.format.extentx, 191 leavesen
dc.format.mediumBTDen
dc.format.mimetypeapplication/pdfen
dc.identifier.otheretd-07282008-134210en
dc.identifier.sourceurlhttp://scholar.lib.vt.edu/theses/available/etd-07282008-134210/en
dc.identifier.urihttp://hdl.handle.net/10919/38851en
dc.language.isoenen
dc.publisherVirginia Techen
dc.relation.haspartLD5655.V856_1992.E237.pdfen
dc.relation.isformatofOCLC# 26064283en
dc.rightsIn Copyrighten
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/en
dc.subject.lccLD5655.V856 1992.E237en
dc.subject.lcshCell suspensionsen
dc.subject.lcshLigninen
dc.subject.lcshLoblolly pineen
dc.subject.lcshPlant cell wallsen
dc.titleCharacterization of lignin deposition in <i>Pinus taeda</i> L. cell suspension culturesen
dc.typeDissertationen
dc.type.dcmitypeTexten
thesis.degree.disciplineWood Science and Forest Productsen
thesis.degree.grantorVirginia Polytechnic Institute and State Universityen
thesis.degree.leveldoctoralen
thesis.degree.namePh. D.en

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