Skip to main content
PMC home page
Plant Physiology logoLink to Plant Physiology
. 1989 Jun;90(2):742–748. doi: 10.1104/pp.90.2.742

Assay and Electrophoresis of Superoxide Dismutase from Red Spruce (Picea rubens Sarg.), Loblolly Pine (Pinus taeda L.), and Scotch Pine (Pinus sylvestris L.) 1

A Method for Biomonitoring

Norman E Tandy 1, Richard T Di Giulio 1, Curtis J Richardson 1
PMCID: PMC1061790  PMID: 16666837

Abstract

This paper reports a method for extracting the antioxidant enzyme superoxide dismutase (SOD) from the needles of red spruce (Picea rubens Sarg.), loblolly pine (Pinus taeda L.), and scotch pine (Pinus sylvestris L.) with high efficiency and free from interfering compounds. The extraction employs phosphate buffer with polyvinylpolypyrrolidone and Triton X-100 followed by dialysis overnight. The isozymes of SOD in each species were separated electrophoretically and tested for their sensitivity to KCN and H2O2. An isozyme resistant to these inhibitors was found in the spruce but not the pine needles. The isozymes from the spruce needles were examined for individual responses to aging and H2O2 inhibition. Four of the five CuZn isozymes in spruce were found to have increased significantly but equally by October of their first year and two of those four isozymes were found to be more sensitive to H2O2. The response of the SOD isozymes in loblolly pine seedlings to O3 was also examined and the isozymes were found to be induced equally. Because the SOD activity in the young pine needles was too low to electrophorese, the SOD activity from the pines in the O3 experiment had to be partially purified using CHCl3 and ethanol, then concentrated.

Full text

PDF
742

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aibara S., Yamashita H., Mori E., Kato M., Morita Y. Isolation and characterization of five neutral isoenzymes of horseradish peroxidase. J Biochem. 1982 Aug;92(2):531–539. doi: 10.1093/oxfordjournals.jbchem.a133961. [DOI] [PubMed] [Google Scholar]
  2. Beauchamp C. O., Fridovich I. Isozymes of superoxide dismutase from wheat germ. Biochim Biophys Acta. 1973 Jul 12;317(1):50–64. doi: 10.1016/0005-2795(73)90198-0. [DOI] [PubMed] [Google Scholar]
  3. Beauchamp C., Fridovich I. Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem. 1971 Nov;44(1):276–287. doi: 10.1016/0003-2697(71)90370-8. [DOI] [PubMed] [Google Scholar]
  4. Bridges S. M., Salin M. L. Distribution of iron-containing superoxide dismutase in vascular plants. Plant Physiol. 1981 Aug;68(2):275–278. doi: 10.1104/pp.68.2.275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Civalleri L., Pini C., Rigo A., Federico R., Calabrese L., Rotilio G. Isolation and preliminary characterization of electrophoretic variants of copper, zinc superoxide dismutase. Mol Cell Biochem. 1982 Aug 20;47(1):3–9. doi: 10.1007/BF00241560. [DOI] [PubMed] [Google Scholar]
  6. Crosti N. On the two electrophoretic forms of the human superoxide dismutase A in the homozygote SOD A1. Biochem Genet. 1978 Aug;16(7-8):739–742. doi: 10.1007/BF00484730. [DOI] [PubMed] [Google Scholar]
  7. Esterbauer H., Grill D. Seasonal Variation of Glutathione and Glutathione Reductase in Needles of Picea abies. Plant Physiol. 1978 Jan;61(1):119–121. doi: 10.1104/pp.61.1.119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Lee E. H., Bennett J. H. Superoxide Dismutase: A POSSIBLE PROTECTIVE ENZYME AGAINST OZONE INJURY IN SNAP BEANS (PHASEOLUS VULGARIS L.). Plant Physiol. 1982 Jun;69(6):1444–1449. doi: 10.1104/pp.69.6.1444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Loomis W. D. Overcoming problems of phenolics and quinones in the isolation of plant enzymes and organelles. Methods Enzymol. 1974;31:528–544. doi: 10.1016/0076-6879(74)31057-9. [DOI] [PubMed] [Google Scholar]
  10. Matters G. L., Scandalios J. G. Effect of the free radical-generating herbicide paraquat on the expression of the superoxide dismutase (Sod) genes in maize. Biochim Biophys Acta. 1986 Jun 3;882(1):29–38. doi: 10.1016/0304-4165(86)90051-6. [DOI] [PubMed] [Google Scholar]
  11. McCord J. M., Fridovich I. Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein). J Biol Chem. 1969 Nov 25;244(22):6049–6055. [PubMed] [Google Scholar]
  12. Mehlhorn H., Seufert G., Schmidt A., Kunert K. J. Effect of SO(2) and O(3) on Production of Antioxidants in Conifers. Plant Physiol. 1986 Sep;82(1):336–338. doi: 10.1104/pp.82.1.336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Salin M. L., Bridges S. M. Isolation and Characterization of an Iron-Containing Superoxide Dismutase From Water Lily, Nuphar luteum. Plant Physiol. 1982 Jan;69(1):161–165. doi: 10.1104/pp.69.1.161. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES