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Dna insertional mutagenesis for the elucidation of a Photosystem II repair process in the green alga Chlamydomonas reinhardtii

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Abstract

The work outlines the isolation of transformant Chlamydomonas reinhardtii cells that appear to be unable to repair Photosystem II from photoinhibitory damage. A physiological and biochemical characterization of three mutants is presented. The results show differential stability for the D1 reaction center protein in the three mutants compared to the wild type and suggest lesions that affect different aspects of the Photosystem II repair mechanism. In the ag16.2 mutant, significantly greater amounts of D1 accumulate in the thylakoid membrane than in the wild type under steady-state growth conditions, and D1 loss is significantly retarded in the presence of the protein biosynthesis inhibitor chloramphenicol. Moreover, aberrant electrophoretic mobility of D1 in the ag16.2 suggests that this protein is modified to an as yet unknown configuration. These results indicate that the biosynthesis and/or degradation of D1 is altered in this strain. A different type of mutation occurred in the kn66.7 and kn27.4 mutants of C. reinhardtii. The stability of D1 declined much faster as a function of light intensity in these mutants than in the wild type. Thereby, the threshold of photoinhibition in these mutants was significantly lower than that in the wild type. It appears that kn66.7 and kn27.4 are similar conditional mutants, with the only difference between them being the amplitude of the chloroplast response to the mutation and the differential sensitivity they display to the level of irradiance.

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Authors and Affiliations

  1. Department of Plant Biology, University of California, Berkeley, CA, 94720-3102, USA

    Liping Zhang, Irene Baroli, Jeff A. Nemson & Anastasios Melis

  2. Department of Plant Biology, Carnegie Institution of Washington, Stanford, CA, 94305-1297, USA

    Krishna K. Niyogi & Arthur R. Grossman

Authors
  1. Liping Zhang
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  2. Krishna K. Niyogi
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  3. Irene Baroli
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  4. Jeff A. Nemson
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  5. Arthur R. Grossman
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  6. Anastasios Melis
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Zhang, L., Niyogi, K.K., Baroli, I. et al. Dna insertional mutagenesis for the elucidation of a Photosystem II repair process in the green alga Chlamydomonas reinhardtii. Photosynthesis Research 53, 173–184 (1997). https://doi.org/10.1023/A:1005867709441

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