The localization of peroxsdase in the cell of several
Spirogyra species was investigated by the aid of benzidine oxidation reaction.
The blue staining with the oxidized benzidine was recognized at first on microsomes, nucleoli and septum regions, and then in whole nuclei.
Plasmolysing the cells with a 1mol. sugar solution before or after the treatment with reagents, it was revealed that the staining of the septum region was, in general, that of the surface of cytoplasm, i.e. the protoplasm membrane. In the
case of certain species of
Spirogyra, however, the cell membrane at septum region, as well as the protoplasm membrane, was stained.
When the cells were treated with the coloured benzidine solution, prepared by
addition of Cl
2-containing tap water to the acidified benzidine solution, the cell membrane was stained blue as stated above, though the microsomes, the nucleoli and the protoplasm membrane remained colourless. The staining of microsomes, nucleoli and protoplasm membrane was inhibited by KCN or boiling (10min.), while that of the cell membrane was not by KCN. Therfore, the staining of the cell membrane of these species is supposed to be caused by the secondary adsorption of the dyes.
In a poorly growing culture, the pathologically isolated septum apparatuses occasionally adsorbed the dyes.
Considering the results of this experiment, it may be concluded with certainty that peroxidase is localized on microsomes, nucleolus and the protoplasm membrane in the cell of
Spirogyra.
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