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Further enzymic studies and electron microscopy of the microbodies of a mutant of Chlorella vulgaris

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Summary

  1. 1.

    The presence of microbodies in the yellow mutant Chlorella vulgaris 211-2-11 h/20 has been demonstrated by means of electron microscopy.

  2. 2.

    A microbody preparation, obtained from the algae by cell disruption and subsequent sucrose density gradient centrifugation, contained an enzyme pattern which suggests that the microbodies from glucose-grown cells are of the peroxisomal rather than of the glyoxysomal type. No malate synthetase and no isocitrate lyase were found.

  3. 3.

    Microbodies of acetate-grown cells also did not contain isocitrate lyase or malate synthetase activities. In contrast to glucose-grown cells a supernatant fraction of acetate-grown cells exhibited low isocitrate lyase activity, but just as in the case of glucose-grown cells activity of malate synthetase was not detectable even if assayed for with a variety of methods, including the use of 14C-1-acetate.

  4. 4.

    In acetate-grown cells malate was the major product of acetate assimilation as evidenced by the characterization of short-term products of 14C-acetate assimilation. Malate was not a primary product of 14C-acetate assimilation in glucose-grown cells.

  5. 5.

    Since on the one hand the presence of malate synthetase could not be demonstrated in acetate-grown cells as well as in glucose-grown cells, on the other hand malate was the primary product of 14C-acetate assimilation in acetate-grown cells it must be concluded, either that the enzyme is so sensitive that it does not survive isolation, or that the mutant utilizes a pathway other than the glyoxylate by-pass for acetate assimilation when dependent upon acetate for growth.

  6. 6.

    As previously demonstrated (Codd et. al., 1972) the isolated microbodies from glucose-grown cells contain high amounts of catalase. An attempt to localize catalase cytochemically by the diaminobenzidine technique failed: diaminobenzidine plus H2O2 stained the microbodies, the mitochondria, Golgi apparatus, and even the lamellar system of the chlorophyll-free plastid. Staining of both the microbodies and mitochondria was relieved by aminotriazole.

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Abbreviations

Aminotriazole:

3-amino-1 H-1,2,4-triazole

DAB:

3,3-diaminobenzidine

DTNB:

5,5′-dithiobis-(2-nitrobenzoic acid)

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Author notes

  1. Geoffrey A. Codd

    Present address: Dept. of Biological Sciences, The University, DD1 4HN, Dundee, Scotland

Authors and Affiliations

  1. Abteilung Menke, Max-Planck-Institut für Züchtungsforschung (Erwin-Baur-Institut), Köln

    Geoffrey A. Codd & Georg H. Schmid

  2. Botanisches Institut der Universität zu Köln, Köln, Germany

    Wolfgang Kowallik

Authors
  1. Geoffrey A. Codd
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  2. Georg H. Schmid
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  3. Wolfgang Kowallik
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Codd, G.A., Schmid, G.H. & Kowallik, W. Further enzymic studies and electron microscopy of the microbodies of a mutant of Chlorella vulgaris . Archiv. Mikrobiol. 92, 21–38 (1973). https://doi.org/10.1007/BF00409508

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