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Generation of active oxygen species during enzymic isolation of protoplasts from oat leaves

  • Plant Cellular and Developmental Biology
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Summary

Protoplasts were isolated from oat (Avena sativa L.) leaves by the combination of highly purified preparations of pectin lyase, xylanase, and cellulase C1. During the enzymic isolation, superoxide radical (O 2− ) was generated from the tissues. Both the protoplasts themselves and the cell walls, exposed to enzyme treatment, produced O 2− . Hydrogen peroxide (H2O2) apparently accumulated in the reaction mixture due to the spontaneous dismutation reaction of O 2− , while a part of H2O2 may have been produced directly from cell walls by the action of enzymes. Singlet molecular oxygen (1O2) generated in the reaction mixture was detected by cholesterol oxidation in small unilamellar liposomes. It seems likely that1O2 may be generated by the peroxidase-H2O2-halide system during enzymic treatment of the leaves.

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Authors and Affiliations

  1. Bioscience Research Laboratory, Kikkoman Corporation, 399 Noda, Noda-shi, Chiba-ken 278, Japan

    Shigetaka Ishii

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  1. Shigetaka Ishii
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The work was partially supported by the Research Project “Research and development of the improvement of bacterial and plant cells by cell fusion” of the Food and Agriculture Research and Development Association (Japan).

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Ishii, S. Generation of active oxygen species during enzymic isolation of protoplasts from oat leaves. In Vitro Cell Dev Biol 23, 653–658 (1987). https://doi.org/10.1007/BF02621075

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  • DOI: https://doi.org/10.1007/BF02621075

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