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PLANT PHYSIOLOGY , Vol 114, Issue 3 1123-1133, Copyright © 1997 by American Society of Plant Biologists
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DEVELOPMENT AND GROWTH REGULATION |
Study of Lignification by Noninvasive Techniques in Growing Maize Internodes (An Investigation by Fourier Transform Infrared Cross-Polarization-Magic Angle Spinning 13C-Nuclear Magnetic Resonance Spectroscopy and Immunocytochemical Transmission Electron Microscopy)
J. P. Joseleau and K. Ruel
Centre de Recherches sur les Macromolecules Vegetales, Centre National de la Recherche Scientifique, Associe a I'Universite Joseph Fourier, BP 53, 38041 Grenoble cedex 9, France
Noninvasive techniques were used for the study in situ of lignification in
the maturing cell walls of the maize (Zea mays L.) stem. Within the
longitudinal axis of a developing internode all of the stages of
lignification can be found. The synthesis of the three types of lignins,
p-hydroxyphenylpropane (H), guaiacyl (G), and syringyl (S), was
investigated in situ by cross-polarization-magic angle spinning
13C-solid-state nuclear magnetic resonance, Fourier transform infrared
spectroscopy, and immunocytochemical electron microscopy. The first lignin
appearing in the parenchyma is of the G-type, preceeding the incorporation
of S nuclei in the later stages. However, in vascular bundles, typical
absorption bands of S nuclei are visible in the Fourier transform infrared
spectra at the earliest stage of lignification. Immunocytochemical
determination of the three types of lignin in transmission electron
microscopy was possible thanks to the use of antisera prepared against
synthetic H, G, and the mixed GS dehydrogenative polymers (K. Ruel, O.
Faix, J.P. Joseleau [1994] J Trace Microprobe Tech 12: 247-265). The
specificity of the immunological probes demonstrated that there are
differences in the relative temporal synthesis of the H, G, and GS lignins
in the different tissues undergoing lignification. Considering the
intermonomeric linkages predominating in the antigens used for the
preparation of the immunological probes, the relative intensities of the
labeling obtained provided, for the first time to our knowledge,
information about the macromolecular nature of lignins (condensed versus
noncondensed) in relation to their ultrastructural localization and
developmental stage.
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