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JXB Advance Access published online on June 13, 2005
Journal of Experimental Botany, doi:10.1093/jxb/eri207
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© The Author [2005]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oupjournals.org
Received December 16, 2004
Accepted April 22, 2005

RESEARCH PAPER

Hydrogen peroxide and expression of hipI-superoxide dismutase are associated with the development of secondary cell walls in Zinnia elegans

Marlene Karlsson 1, Michael Melzer 2, Isabella Prokhorenko 3, Thorsten Johansson 4, and Gunnar Wingsle 1*

1 Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Science, Umeå Plant Science Centre, 90183 Umeå, Sweden
2 Department of Molecular Cell Biology, Institute of Plant Genetics and Crop Plant Research, 06466 Gatersleben, Germany
3 Institute of Basic Biological Problems, Russian Academy of Sciences, Pushtchino, Moscow Region, 142292 Russia
4 Swedish Defense Research Agency, FOI, NBC, Defense Department of Threat Assessment, 901 82 Umeå, Sweden

* To whom correspondence should be addressed.
Gunnar Wingsle, E-mail: gunnar.wingsle{at}genfys.slu.se


  Abstract

A special form of a CuZn-superoxide dismutase with a high isoelectric point (hipI-SOD; EC 1.15.1.1) and hydrogen peroxide (H2O2) production were studied during the secondary cell wall formation of the inducible tracheary element cell-culture system of Zinnia elegans L. Confocal microscopy after labelling with 2',7'-dichlorofluorescin diacetate showed H2O2 to be located largely in the secondary cell walls in developing tracheary elements. Fluorescence-activated cell sorting analysis showed there were lower levels of H2O2 in the population containing tracheary elements when H2O2 scavengers such as ascorbate, catalase, and reduced glutathione were applied to the cell culture. Inhibitors of NADPH oxidase and SOD also reduced the amount of H2O2 in the tracheary elements. Furthermore, addition of these compounds to cell cultures at the time of tracheary element initiation reduced the amount of lignin and the development of the secondary cell walls. Analysis of UV excitation under a confocal laser scanning microscope confirmed these results. The expression of hipI-SOD increased as the number of tracheary elements in the cell culture increased and developed. Additionally, immunolocalization of a hipI-SOD isoform during the tracheary element differentiation showed a developmental build-up of the protein in the Golgi apparatus and the secondary cell wall. These findings suggest a novel hipI-SOD could be involved in the regulation of H2O2 required for the development of the secondary cell walls of tracheary elements.

Keywords: CuZn-superoxide dismutase; hydrogen peroxide; immunolocalization; tracheary element; Zinnia cell-culture system.
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