Plant Physiol.
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Plant Physiology 89:786-791 (1989)
© 1989 American Society of Plant Biologists

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Molecular Biology and Gene Regulation

Characterization of Nitrate Reductase Deficient Mutants of Chlorella sorokiniana

Otto Knobloch and Rudolf Tischner

Institut für Pflanzenphysiologie, University of Göttingen, Untere Karspüle 2, 3400 Göttingen, West Germany

After x-ray irradiation, 13 mutants of Chlorella sorokiniana incapable of using NO3– as N source were isolated using a pinpoint method. Using immunoprecipitation and Western blot assays, no nitrate reductase was found in five strains while in eight mutants the enzyme was detected. The latter strains contained different patterns of nitrate reductase partial reactions. All isolates were of the nia-type as indicated by the inducibility of purine hydroxylase I and by complementation of nitrate reductase activity in the Neurospora crassa mutant Nit-1. A restoration of NADP-nitrate reductase in Nit-1 was also obtained with NH4+-grown cells indicating that Mo-cofactor is constitutive in Chlorella. Complementation experiments among the Chlorella mutants resulted in restoration of NADH-nitrate reductase activity. The characteristics of some of the Chlorella mutants are discussed in view of an improper orientation of Mo-cofactor in the residual nitrate reductase protein.






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Copyright © 1989 by the American Society of Plant Biologists