Advertisement

Molecular and General Genetics MGG

, Volume 181, Issue 1, pp 20–23 | Cite as

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

  • T. Kuo
  • A. Kleinhofs
  • D. Somers
  • R. L. Warner
Article

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.

Keywords

Nitrate Reductase Hordeum Vulgare Rabbit Antibody Protection Assay Diffusion Test 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Abbreviations

NR

nitrate reductase

DTT

dithiothreitol

FAD

flavin adenine dinucleotide

BSA

bovine serum albumin

NRCRM

nitrate reductase cross-reacting materials

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Amy NK, Garrett RH (1979) Immunoelectrophoretic determination of nitrate reductase in Neurospora crassa. Anal Biochem 95:97–107Google Scholar
  2. Arst HN, MacDonald DW, Cove DJ (1970) Molybdate metabolism in Aspergillus nidulans. I. Mutations affecting nitrate reductase and/or xanthine dehydrogenase. Mol Gen Genet 108:129–145Google Scholar
  3. Cove DJ (1979) Genetic studies of nitrate assimilation in Aspergillus nidulans. Biol Rev 54:291–327Google Scholar
  4. Crowle AJ, Cline LJ (1977) An improved stain for immunodiffusion tests. J Immunol Meth 17:379–381Google Scholar
  5. Funkhouser EA, Ramadoss CS (1980) Synthesis of nitrate reductase in Chlorella II. Evidence for synthesis in ammonia-grown cells. Plant Physiol 65:944–948Google Scholar
  6. Garrett RH, Amy NK (1978) Nitrate assimilation in fungi. Adv Microbiol Physiol 18:1–65Google Scholar
  7. Graf L, Notton BA, Hewitt EJ (1975a) Serological estimation of spinach nitrate reductase. Phytochemistry 14:1241–1243Google Scholar
  8. Graf L, Notton BA, Hewitt EJ (1975b) The effects of immune and non-immune rabbit sera on the various activities of spinach (Spinacea oleracea L.) nitrate reductase. Plant Sci Lett 4:69–75Google Scholar
  9. Kleinhofs A, Kuo T, Warner RL (1980) Characterization of nitrate reductase-deficient barley mutants. Mol Gen Genet 177:421–425Google Scholar
  10. Kleinhofs A, Warner RL, Muehlbauer FJ, Nilan RA (1978) Induction and selection of specific gene mutations in Hordeum and Pisum. Mutation Res 51:29–35Google Scholar
  11. Kuo T (1979) Nitrate reductase inbarley: Nature and biochemical characterization of mutants. Dissertation, Washington State University, PullmanGoogle Scholar
  12. Kuo T, Kleinhofs A, Warner RL (1979) Antigenic properties of nitrale reductase-deficient mutants in Hordeum vulgare L. Genetics 91: s64Google Scholar
  13. Kuo T, Kleinhofs A, Warner RL (1980) Purification and partial characterization of nitrate reductase frombarley leaves. plant Sci Lett 17:371–381Google Scholar
  14. Loomis WD (1974) Overcoming problems of phenolies and quainones in the isolation of plant enzymes and organelles. Methods Enzymol 31:528–544Google Scholar
  15. Macdonald DW, Cove DJ, Coddington A (1974) Cytochrome-C reductase from wild-type and mutant strains of Aspergillus nidulans. Mol Gen Genet 128:187–199Google Scholar
  16. Medel RR, Müller AJ (1978) Reconstitution of NADH-nittrate reductase in vitro from nitrate reductase-deficient Nicotiana tabacum mutants. Mol Gen Genet 161:77–80Google Scholar
  17. Mendel RR, Müller AJ (1979) Nitrate reductase-deficient mutant cell lines of Nicotiana tabacum. Mol Gen Genet 177:145–153Google Scholar
  18. Mendel RR, Müller AJ (1980) Comparative characterization of nitrate reductase from wild-type and molybedenum cofactor-defective cell cultures of Nicotiana tabacum. PlantSci Lett 18:277–288Google Scholar
  19. Müller AJ, Grafe R (1978) Isolation and characterization of cell lines of Nicotiana tabacum lacking nitrate reductase. Mol Gen Genet 161:67–76Google Scholar
  20. Murray ED, Sanwal BD (1963) An immunological enquiry into the identity of assimilatory and dissimilatory nitrate reductase from Escherichia coli. Can J Microbiol 9:781–790Google Scholar
  21. Notton BA, Graf L, Hewitt EJ, Povey RC (1974) The role of molybdenum in the synthesis of nitrate reductase in cauliflower (Brassicaoleracea L. var. Botrytis L.) and spinach (Spinacea oleracea L.). Biochim BIophys Acta 364:45–58Google Scholar
  22. Oostindier-Braaksma FJ, Feenstra WJ (1973) Isolation and characterization of chlorate-resistant mutants of Arabidopsis thaliana. Muttion Res 19:175–185Google Scholar
  23. Ouchterlony O (1949) antigen-antibody reaction in gels. Acta Pathol Microbiol Scand 26:507–515Google Scholar
  24. Pateman JA, Cove DJ, Rever BM, roberts DB (1964) common co-factor for nitrate reductase and xanthine dehydrogenase which alos aregulates the synthesis of nitrate reductase. Nature 201:58–60Google Scholar
  25. Scholl RL, Harper JE, Hageman RH (1974) Improvement of the nitrite color development in assays of nitrate reductase by phenzatine mthosulfate and zinc acetate. Plant Physiol 53:825–828Google Scholar
  26. siegel NR, Enns RK (1979) soluble polyvinylpyrrolidine and bovine serum albumin adsorb polyphenods from soybean suspension cultures. Plant Physiol 63:206–208Google Scholar
  27. Sorger GJ (1966) Nitrate reductase electron transport systems in mutant and in wild type strains of Neurospora. Biochim Biocphys Acta 118:484–494Google Scholar
  28. Sorger GJ, Davies J (1973) Regulation of nitrate reductase of Neurospora at the level of transcription and translation. Biochem j 134:673–685Google Scholar
  29. sorger GJ, Debanne MT, Davies J (1974) Effect of nitrate on the synthesis and decay of nitrate reductase of Neurospora. Biochem J 140:395–403Google Scholar
  30. Sorger GJ, giles NH (1965) Genetic control of nitrate reductase in Neurospora crassa. Genetics 52:777–788Google Scholar
  31. tokarev BI, Shumny VK (1977) detection of barley mutants with low level of nitrate reductase activity after the seed treatment with ethylmthanesulphonate. Genetika Moskva 13:2097–2103Google Scholar
  32. Warner RL, Kleinhofs A (1974) Relationships between nitrate reductase, nitrite reductase, and ribulose diphosphate carboxylase activities in chlorophyll-deficienty mutants ofbarley. Crop Sci 14:654–658Google Scholar
  33. Warner RL, Lin CJ, Kleinhofs A (1977) Nitrate reductase-deficient mutants in barley. Natue 269:406–407Google Scholar

Copyright information

© Springer-Verlag 1981

Authors and Affiliations

  • T. Kuo
    • 1
  • A. Kleinhofs
    • 1
  • D. Somers
    • 2
  • R. L. Warner
    • 2
  1. 1.Department of Agronomy and Soils and Program in GeneticsWashington State UniversityPullmanUSA
  2. 2.Department of Agronomy and SoilsWashington State UniversityPullmanUSA

Personalised recommendations