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Planta

, Volume 201, Issue 3, pp 311–318 | Cite as

Fourier-transform infrared and Raman spectroscopic evidence for the incorporation of cinnamaldehydes into the lignin of transgenic tobacco (Nicotiana tabacum L.) plants with reduced expression of cinnamyl alcohol dehydrogenase

  • Derek Stewart
  • Nabila Yahiaoui
  • Gordon J. McDougall
  • Kate Myton
  • Christiané Marque
  • Alain M. Boudet
  • James Haigh
Article

Abstract

Xylem from stems of genetically manipulated tobacco plants which had had cinnamyl alcohol dehydrogenase (CAD; EC 1.1.1.195) activity down-regulated to a greater or lesser degree (clones 37 and 49, respectively) by the insertion of antisense CAD cDNA had similar, or slightly higher, lignin contents than xylem from wild-type plants. Fourier-transform infrared (FT-IR) microspectroscopy indicated that down-regulation of CAD had resulted in the incorporation of moieties with conjugated carbonyl groups into lignin and that the overall extent of cross-linking, particularly of guaiacyl (4-hydroxy-3-methoxyphenyl) rings, in the lignin had altered. The FT-Raman spectra of manipulated xylem exhibited maxima consistent with the presence of elevated levels of aldehydic groups conjugated to a carbon-carbon double bond and a guaiacyl ring. These maxima were particularly intense in the spectra of xylem from clone 37, the xylem of which exhibits a uniform red coloration, and their absolute frequencies matched those of coniferaldehyde. Furthermore, xylem from clone 37 was found to have a higher content of carbonyl groups than that of clone 49 or the wild-type (clone 37: clone 49: wild-type; 2.4:1.6:1.0) as measured by a degradative chemical method. This is the first report of the combined use of FT-IR and FT-Raman spectroscopies to study lignin structure in situ. These analyses provide strong evidence for the incorporation of cinnamaldehyde groups into the lignin of transgenic plants with down-regulated CAD expression. In addition, these non-destructive analyses also suggest that the plants transformed with antisense CAD, in particular clone 37, may contain lignin that is less condensed (cross-linked) than that of the wild-type.

Key words

Lignin-Cell walls Cinnamyl alcohol dehydrogenase Infrared Raman 

Abbreviations

CAD

cinnamyl alcohol dehydrogenase

FT-IR

Fourier-transform infrared

DRIFT

diffuse reflectance Fourier-transform infrared

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Copyright information

© Springer-Verlag 1997

Authors and Affiliations

  • Derek Stewart
    • 1
  • Nabila Yahiaoui
    • 2
  • Gordon J. McDougall
    • 1
  • Kate Myton
    • 2
  • Christiané Marque
    • 2
  • Alain M. Boudet
    • 2
  • James Haigh
    • 3
  1. 1.Unit for Industrial Crops, Department of Cellular and Environmental PhysiologyScottish Crop Research InstituteDundeeUK
  2. 2.Unité Mixte de Recherche UPS-CNRS No 5546University Paul SabatierToulouse CedexFrance
  3. 3.Department of ChemistryUniversity of SouthamptonSouthamptonUK

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