Plant Physiol. Journal of Pharmacology and Experimental Therapeutics
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Plant Physiology 77:232-236 (1985)
© 1985 American Society of Plant Biologists

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Articles

Immunochemical Characterization of Nitrate Reductase Forms from Wild-Type (cv Williams) and nr1 Mutant Soybean 1

Paul Robin2, Luc Streit, Wilbur H. Campbell and James E. Harper

Department of Chemistry State University of New York, College of Environmental Science and Forestry, Syracuse, New York 13210, Department of Agronomy, University of Illinois, Urbana, Illinois 61801, United States Department of Agriculture, Agriculture Research Service, University of Illinois, Urbana, Illinois 61801

Soybean (Glycine max L. Merr.) leaves contain two forms of nitrate reductase (NR)—NAD(P)H:NR and NADH:NR. Wild-type (cv Williams), nr1 mutant and an unrelated cultivar (Prize) were grown with either no N source or with nitrate. Crude extracts were assayed for NR activities and the enzyme forms were purified on blue Sepharose. Analyses were done by polyacrylamide gel electrophoresis and `Western blotting' using antibodies specific for NR. NAD(P)H:NR was identified as the constitutive NR present in wild-type and Prize, but was absent from the mutant. All three soybean lines contained nitrate-inducible NADH:NR with highest activity at pH 7.5. The results showed that NAD(P)H:NR and constitutive NR were one in the same and confirmed the presence of NADH:NR with pH 7.5 optimum.

2 On leave from Laboratoire de Biochemie et Physiologie Vegetale, Institut National de la Recherche Agronomique, 34060 Montpellier CEDEX, France, and partly supported by NATO.

1 Supported by National Science Foundation Grant (PCM 83-02146) and United States Department of Agriculture Competitive Research Grant (83-CRCR-1-1289) to W.H.C.; by United States Department of Agriculture Competitive Research Grant (82-CRCR-1-1038) to J.E.H.; and by partial support to L.S. from Swiss National Foundation.






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Copyright © 1985 by the American Society of Plant Biologists