Antigenicity of nitrate reductase-deficient mutants in Hordeum vulgare L.

Summary Ten nitrate reductase (NR)-deficient mutants have been characterized for their cross-reactivity against specific barley (Hordeum vulgare L.) nitrate reductase antibodies. The rabbit antibodies raised against the purified barley wild type (cv. Steptoe) enzyme quantitatively inactivate nitrate reductase in crude extracts. All nitrate-grown (induced) mutants show positive precipitin reaction against the antiserum by Ouchterlony double diffusion test and all have the ability to neutralize antisera in a NR protection assay. Under induced growth conditions, mutants Az 12, Az 23, Az 29 and Az 30 which have low NR associated catalytic activities also have the lowest level of antigenicity; mutants Az 13, Az 31, Az 33 and Az 34 have intermediate level of both NR associated catalytic activities and antigenicity, while mutants Az 28 and Az 32 have the highest level of both NR associated catalytic activities and antigenicity. Under noninduced growth conditions, all mutants except Az 12 contain detectable but very low levels of NR antigenicity. These results support the concept that these NR-deficient mutants with various levels of NR associated catalytic activities represent different mutation events at the loci coding the NR structural components.Abbreviations NR nitrate reductase - DTT dithiothreitol - FAD flavin adenine dinucleotide - BSA bovine serum albumin - NRCRM nitrate reductase cross-reacting materialsScientific Paper No. 5765. College of Agriculture Research Center, Washington State University, Pullman, Project Nos. 0233 and 0430. Supported in part by National Science Foundation Grant #PCM7807649, and U.S. Department of Agriculture CRGO Grant #7900536