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Protoplasma

, Volume 250, Issue 2, pp 623–629 | Cite as

The Tr-cp 14 cysteine protease in white clover (Trifolium repens) is localized to the endoplasmic reticulum and is associated with programmed cell death during development of tracheary elements

  • Maria Mulisch
  • Torben AspEmail author
  • Karin Krupinska
  • Julien Hollmann
  • Preben Bach Holm
Short Communication

Abstract

Cysteine proteases are known to be associated with programmed cell death, developmental senescence and some types of pathogen and stress-induced responses. In the present study, we have characterized the cysteine protease Tr-cp 14 in white clover (Trifolium repens). Tr-cp 14 belongs to the C1A family of cysteine proteases with homology to XCP1 and XCP2 from Arabidopsis thaliana and p48h-17 from Zinnia elegans, which previously have been reported to be associated with tracheary element differentiation. The proform as well as the processed form of the protein was detected in petioles, flowers and leaves, but the processed form was more abundant in leaves and petioles than in flowers. The Tr-cp 14 protein was localized to differentiating tracheary elements within the xylem, indicating that the cysteine protease is involved in protein re-mobilization during tracheary element differentiation. Immunogold studies suggest that the protease prior to the burst of the vacuole was associated to the ER cisternae. After disruption of the tonoplast, it was found in the cytoplasm, and, in later stages, associated with disintegrating material dispersed throughout the cell.

Keywords

Cysteine protease Endoplasmic reticulum Immunolocalization Tracheary element differentiation Trifolium repens 

Notes

Acknowledgments

Sanne Seval and Ole Braad Hansen, Aarhus University, are acknowledged for technical and horticultural assistance, respectively. Anke Schäfer, University of Kiel, is acknowledged for purification of the antibody. Marita Beese is acknowledged for assistance in the electron microscopical preparations. The project was supported by the Danish Food Industry Agency through the programme “Biotechnology in Food Research”.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Maria Mulisch
    • 2
  • Torben Asp
    • 1
    Email author
  • Karin Krupinska
    • 2
  • Julien Hollmann
    • 2
  • Preben Bach Holm
    • 1
  1. 1.Faculty of Science and Technology, Department of Molecular Biology and Genetics, Research Centre FlakkebjergAarhus UniversitySlagelseDenmark
  2. 2.Institute of Botany and Central MicroscopyChristian-Albrechts-University KielKielGermany

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