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Related Experiment Videos

Superoxide dismutase activity in root-colonizing pseudomonads

J Katsuwon1, R Zdor, A J Anderson

  • 1Biology Department, Utah State University, Logan 84322-5305.

Canadian Journal of Microbiology
|April 1, 1993
PubMed
Summary

Saprophytic fluorescent pseudomonads produce superoxide dismutase (SOD) isoforms, with the hydrogen peroxide insensitive form influenced by iron and manganese. This SOD activity is crucial for root colonization by these bacteria.

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Area of Science:

  • Microbial Physiology
  • Plant-Microbe Interactions
  • Enzymology

Background:

  • Saprophytic fluorescent pseudomonads are aggressive root colonizers.
  • Superoxide dismutase (SOD) plays a role in bacterial defense and survival.
  • SOD exists in different isoforms, some sensitive and some insensitive to hydrogen peroxide.

Purpose of the Study:

  • To investigate the characteristics and regulation of superoxide dismutase (SOD) isoforms in root-colonizing pseudomonads.
  • To determine the role of SOD in the interaction between pseudomonads and plant roots.
  • To compare SOD expression patterns in Pseudomonas putida with Pseudomonas aeruginosa.

Main Methods:

  • Growth of pseudomonads in liquid culture and analysis of SOD activity and isoforms using nondenaturing polyacrylamide gel electrophoresis.

Related Experiment Videos

  • Investigating the effects of iron availability, Mn2+, and superoxide anion on SOD synthesis.
  • Complementation studies using a plasmid in a superoxide dismutase-deficient Escherichia coli mutant.
  • Assessing bacterial survival and root surface enzyme activity in the presence of plant roots.

    • Main Results:

    • Root-colonizing pseudomonads exhibit similar specific SOD activities in liquid culture.
    • Two SOD isoforms (hydrogen peroxide sensitive and insensitive) with distinct electrophoretic mobilities were identified.
    • Synthesis of the hydrogen peroxide insensitive SOD isoform is enhanced by low iron, Mn2+, and superoxide anion.
    • Unlike P. aeruginosa, P. putida did not show phosphate-regulated SOD isoform patterns.
    • A plasmid encoding the hydrogen peroxide sensitive SOD complemented an E. coli mutant.
    • Bacterial survival on bean roots was not influenced by SOD expression or deficiency.
    • Only the hydrogen peroxide sensitive SOD isoform was detected on bean root surfaces by P. putida.

    Conclusions:

    • Pseudomonads possess distinct SOD isoforms with differential regulation, particularly the hydrogen peroxide insensitive form under specific environmental conditions.
    • The hydrogen peroxide sensitive SOD isoform is important for P. putida colonization of plant roots.
    • SOD expression does not affect bacterial survival on the root surface but is critical for root colonization.