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Related Experiment Videos

Three differentially expressed basic peroxidases from wound-lignifying Asparagus officinalis.

Kirsten B Holm1, Per H Andreasen, Reinhard M G Eckloff

  • 1Plant Research Department, PRD-301, Risø National Laboratory, PO Box 49, DK-4000 Roskilde, Denmark.

Journal of Experimental Botany
|August 30, 2003
PubMed
Summary

Post-harvest, asparagus spears show increased peroxidase activity due to new isoperoxidases, particularly in vascular bundles. Gene expression analysis identified three new class III plant peroxidases, with some showing increased post-harvest activity.

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Area of Science:

  • Plant biochemistry
  • Molecular biology
  • Post-harvest physiology

Background:

  • Peroxidases play crucial roles in plant cell wall metabolism and defense.
  • Understanding post-harvest changes in enzyme activity is vital for maintaining produce quality.
  • Asparagus peroxidases have not been extensively characterized at the molecular level post-harvest.

Purpose of the Study:

  • To investigate the changes in peroxidase activity and identify specific isoperoxidases in asparagus spears post-harvest.
  • To clone and characterize new peroxidase genes from asparagus.
  • To correlate peroxidase activity and localization with tissue changes during post-harvest storage.

Main Methods:

  • Enzyme activity assays and isoelectric focusing to analyze peroxidase isoforms.

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  • Histochemical staining to determine the localization of peroxidase and lignin.
  • cDNA library screening and sequencing to identify peroxidase genes.
  • Gene expression analysis to study peroxidase gene regulation.

    • Main Results:

    • Total peroxidase activity in asparagus spears increased from 5 to 24 hours post-harvest.
    • This increase was attributed to the enhanced activity of several isoperoxidases, with some appearing only at 24 hours.
    • Lignin and peroxidase activity were localized in the vascular bundles, specifically in the protoxylem cell walls.
    • Three new class III plant peroxidase genes were isolated, showing high amino acid sequence identity (70-81%) among themselves.
    • Two of the identified peroxidase genes exhibited increased expression post-harvest.

    Conclusions:

    • Post-harvest storage induces significant changes in asparagus peroxidase activity and expression.
    • The identified class III plant peroxidases are likely involved in the observed post-harvest changes in asparagus spears.
    • Localization studies suggest a role for peroxidases and lignin in structural modifications within vascular tissues during storage.