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Localization of dibenzodioxocin substructures in lignifying Norway spruce xylem by transmission electron microscopy–immunogold labeling

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Abstract

The lignification process in mature Norway spruce [Picea abies (L.) H. Karsten] xylem cell walls was studied using transmission electron microscopy (TEM)–immunogold detection with a polyclonal antibody raised against a specific lignin substructure, dibenzodioxocin. The study reveals for the first time the exact location of this abundant eight-ring structure in the cell wall layers of wood. Spruce wood samples were collected in Southern Finland at the time of active growth and lignification of the xylem cell walls. In very young tracheids where secondary cell wall layers were not yet formed, the presence of the dibenzodioxocin structure could not be shown at all. During secondary cell wall thickening, the dibenzodioxocin structure was more abundant in the secondary cell wall layers than in the middle lamella. The highest number of gold particles revealing dibenzodioxocin was in the S2+S3 layer. Statistically significant differences were found in the frequency of gold particles present in various cell wall layers. For comparison, wood sections were also cut with a cryomicrotome for light and fluorescence microscopy.

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Fig. 1. a
Fig. a–f.
Fig. 3a–f.

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Abbreviations

BSA:

bovine serum albumin

ELISA:

enzyme linked immunosorbent assay

KLH:

keyhole limpet hemocyanin

TEM:

transmission electron microscopy

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Acknowledgements

This investigation was funded by the Academy of Finland (grant no: 43091) and the National Technology Agency TEKES under The Finnish Forest Cluster Research Program WOOD WISDOM. This work is a part of the Center of Excellence in Plant Biology and Forest Biotechnology (project no. 164346) as granted by the Academy of Finland and the Ministry of Education of Finland. Wood samples were prepared and electron micrographs taken at the Electron Microscopy Unit of the Institute of Biotechnology, University of Helsinki.

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Authors and Affiliations

  1. Department of Biosciences, Division of Plant Physiology, Helsinki University, PO Box 56, 00014, Helsinki, Finland

    Eija M. Kukkola & Kurt V. Fagerstedt

  2. Department of Applied Biology, Helsinki University, PO Box 27, 00014, Helsinki, Finland

    Sanna Koutaniemi & Teemu H. Teeri

  3. Department of Chemistry, Laboratory of Organic Chemistry, Helsinki University, PO Box 55, 00014, Helsinki, Finland

    Mikaela Gustafsson, Pirkko Karhunen & Gösta Brunow

  4. Centre de Recherches sur les Macromolécules Végétales, CNRS, BP53, 38041, Grenoble, Cedex 9, France

    Katia Ruel

  5. Department of Applied Chemistry and Microbiology, Helsinki University, PO Box 56, 00014, Helsinki, Finland

    Taina K. Lundell

  6. Finnish Forest Research Institute, PO Box 18, 01301, Vantaa, Finland

    Pekka Saranpää

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  1. Eija M. Kukkola
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  2. Sanna Koutaniemi
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  3. Mikaela Gustafsson
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  8. Gösta Brunow
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  9. Teemu H. Teeri
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  10. Kurt V. Fagerstedt
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Correspondence to Kurt V. Fagerstedt.

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Kukkola, E.M., Koutaniemi, S., Gustafsson, M. et al. Localization of dibenzodioxocin substructures in lignifying Norway spruce xylem by transmission electron microscopy–immunogold labeling. Planta 217, 229–237 (2003). https://doi.org/10.1007/s00425-003-0983-x

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  • DOI: https://doi.org/10.1007/s00425-003-0983-x

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