EPR evidence for generation of hydroxyl radical triggered by N-acetylchitooligosaccharide elicitor and a protein phosphatase inhibitor in suspension-cultured rice cells

  • Kuchitsu K
  • Kosaka H
  • Shiga T
 et al. 
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Abstract

N-acetylchitooligosaccharides, fragments of the backbone of fungal cell wall, trigger rapid membrane responses such as transient depolarization, and elicit defense reactions including phytoalexin production in suspension-cultured rice cells. The generation of reactive oxygen species triggered by the oligosaccharide signal was analyzed with EPR spectroscopy using a spin trapping system, 4-pyridyl 1-oxideN-tert-butyl nitrone (4-POBN) and ethanol. OH generation was detected as the α-hydroxyethyl adduct of 4-POBN after elicitation. Superoxide dismutase, catalase or diethylenetriamine pentaacetic acid, a metal chelator, inhibited TeX{}^ \cdot O generation, proposing the following reaction sequence: generation of TeX{}^ \cdot O_{2^ - } in response to the oligosaccharide elicitor, followed by dismutation to H2O2, then generation of TeX{}^ \cdot OH by the reaction of H2O2 with Fe2+ that is generated by the reduction of Fe3+ by TeX{}^ \cdot O_{2^ - } . Generation of the same reactive oxygen species was also triggered by calyculin A, a protein phosphatase inhibitor, alone, suggesting the involvement of protein phosphorylation in its regulation during the oligosaccharide signal transduction.

Author-supplied keywords

  • Chitin fragments
  • Electron paramagnetic resonance
  • Elicitor
  • Hydroxyl radical
  • Reactive oxygen species
  • Rice suspension culture

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Authors

  • Kazuyuki KuchitsuTokyo University of Science, Department of Applied Biological Science

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  • H. Kosaka

  • T. Shiga

  • N. Shibuya

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