Skip to main content
SpringerLink
Log in

Nitrate reductase activity in Paul's scarlet rose suspension cultures and the differential role of nitrate and molybdenum in induction

  • Published:
Planta Aims and scope Submit manuscript

Abstract

Induction of nitrate reductase (EC 1.6.6.1) activity was measured in Paul's Scarlet rose cell suspensions cultured in media containing nitrate (NO -3 ) or urea (U) as nitrogen source, and with (+Mo) or without molybdenum (-Mo). There was a lag of 30 min during induction by NO -3 in +Mo cultures but no lag occurred during induction after adding Mo to NO -3 -Mo or to U-Mo cultures preincubated with NO -3 . Actinomycin D, cycloheximide, and puromycin completely blocked induction by NO -3 , but had no effect on the initial rate of induction by Mo. Cycloheximide and puromycin blocked induction by NO -3 more quickly than actinomycin D. Induction by NO -3 appeared to involve mRNA-dependent synthesis of apoprotein followed by rapid activation with molybdenum in intact cells independently of protein synthesis. Nitrate-induced apoprotein appeared less stable than the holoenzyme. When induced by NO -3 in the absence of Mo, apoprotein concentration was about half the amount of maximally induced nitrate reductase. Cycloheximide stabilised preformed nitrate reductase which disappeared steadily in the presence of puromycin. Apoprotein was not stabilised by either antimetabolite.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

Mo:

molybdenum

NO -3 +Mo:

standard, MX1 culture medium

NO -3 -Mo:

MX1 medium purified of Mo

NR:

nitrate reductase

PSR:

Paul's Scarlet rose

U:

urea

U+Mo:

MX1 medium with NO -3 replaced by urea

U-Mo:

MX1 medium with NO -3 replaced by urea and also purified of Mo

References

  • Afridi, M.M.R.K., Hewitt, E.J.: The inducible formation and stability of nitrate reductase in higher plants. I. Effects of nitrate and molybdenum on enzyme activity in cauliflower (Brass. oleracea var. Botrytis). J. exp. Bot. 15, 251–271 (1964)

    Google Scholar 

  • Afridi, M.M.R.K., Hewitt, E.J.: The inducible formation and stability of nitrate reductase in higher plants. II. Effects of environmental factors, antimetabolites, and amino acids on induction. J. exp. Bot. 16, 628–645 (1965)

    Google Scholar 

  • Beevers, L., Hageman, R.H.: Nitrate reduction in higher plants. Ann. Rev. Plant Physiol. 20, 495–522 (1969)

    Article  Google Scholar 

  • Behrend, J., Mateles, R.I.: Nitrogen metabolism in plant cell suspension cultures. I. Effect of amino acids on growth. Plant Physiol. 56, 584–589 (1975)

    Google Scholar 

  • Gale, E.F., Cundliffe, E., Reynold, P.E., Richmond, M.H., Waring, M.J.: In: The molecular basis of antibiotic action. London: Wiley 1972

    Google Scholar 

  • Heimer, Y.M., Filner, P.: Regulation of nitrate assimilation pathway in cultured tobacco cells. III. The nitrate uptake system. Biochim. Biophys. Acta 230, 362–372 (1971)

    PubMed  Google Scholar 

  • Hewitt, E.J.: Some aspects of micronutrient element metabolism in plants. Nature 180, 1020 (1957)

    PubMed  Google Scholar 

  • Hewitt, E.J.: The role of mineral elements in the activity of plant enzyme systems. In: Handbook of plant physiology, vol. 4, pp. 427–481. Ruhland, W., ed. Berlin:Springer 1958

    Google Scholar 

  • Hewitt, E.J.: The metabolism of micronutrient elements in plants. Biol. Rev. 34, 333–377 (1959)

    Google Scholar 

  • Hewitt, E.J.: Assimilatory nitrate-nitrite reduction. Ann. Rev. Plant Physiol. 26, 73–100 (1975)

    Google Scholar 

  • Hewitt, E.J., Afridi, M.M.R.K.: Adaptive synthesis of nitrate reductase in higher plants. Nature 183, 57–58 (1959)

    Google Scholar 

  • Hewitt, E.J. Jones, R.W., Abbott, A.J., Best, G.R.: Effect of nitrate and molybdenum on in vivo formation of nitrate reductase in Paul's Scarlet rose cells. Tulecke strain. Abstr. no. 131. 3rd International Congress in Plant Tissue Culture, University of Leicester, England, 1974

  • Hewitt, E.J., Nicholas, D.J.D.: Enzymes of inorganic nitrogen metabolism. In: Modern methods of plant analysis, vol. 7, pp. 67–122. Linskens, H.F., Sanwal, B.D., Tracey, M.V., eds., Berlin: Springer 1964

    Google Scholar 

  • Hewitt, E.J., Notton, B.A.: Inhibition by L-azetidine-2-carboxylic acid of induction of nitrate reductase in plants and its reversal by L-proline. Phytochem. 6, 1329–1335 (1967)

    Google Scholar 

  • Hewitt, E.J., Notton, B.A., Afridi, M.M.R.K.: Comparative effects of some antimetabolites on ribonucleic acid synthesis and induction of nitrate reductase in cauliflower leaf tissue. Plant Cell Physiol. 8, 385–397 (1967)

    Google Scholar 

  • Hewitt, E.J., Notton, B.A., Rucklidge, G.J.: Formation of nitrate reductase by recombination of apoprotein fractions from molybdenum-deficient plants with a molybdenum containing complex. J. Less-Common Metals 54, 537–553 (1977)

    Google Scholar 

  • Ketchum, P.A., Downey, R.J.: In vitro restoration of nitrate reductase: investigation of Aspergillus nidulans and Neurospora crassa nitrate reductase mutants. Biochim. Biophys. Acta 385, 354–361 (1975)

    PubMed  Google Scholar 

  • Jackson, W.A., Flesher, D., Hageman, R.H.: Nitrate uptake by dark-grown corn seedlings, some characteristics of apparent induction. Plant Physiol. 51, 120–127 (1973)

    Google Scholar 

  • Jones, R.W., Abbott, A.J., Hewitt, E.J., Best, G.R., James, D.M.: Nitrate reductase activity and growth in Paul's Scarlet rose suspension cultures in relation to nitrogen source and molybdenum. Planta 133, 27–34 (1976)

    Google Scholar 

  • King, J.: The uptake of nitrate by sycamore cells, Abstr. no. 133. 3rd International Congress on Plant Tissue Culture, University of Leicester, England, 1974

  • Neyra, C.A., Hageman, R.H.: Nitrate uptake and induction of nitrate reductase in excised corn roots. Plant Physiol. 56, 692–695 (1975)

    Google Scholar 

  • Notton, B.A., Graf, L., Hewitt, E.J., Povey, R.C.: The role of molydbenum in the synthesis of nitrate reductase in cauliflower (Brassica oleracea L. var. Botrytis L.) and spinach (Spinacea oleracea L.). Biochim. Biophys. Acta 364, 45–58 (1974)

    PubMed  Google Scholar 

  • Oaks, A., Wallace, W., Stevens, D.: Synthesis and turnover of nitrate reductase in corn roots. Plant Physiol. 50, 649–654 (1972)

    Google Scholar 

  • Rucklidge, G.J., Hewitt, E.J.: Rep. Long Ashton Res. Stat. for 1974. pp. 66–67 (1975)

  • Rucklidge, G., Notton, B., Hewitt, E.: Reconstitution in vitro of nitrate reductase from apoprotein of molybdenum-deficient spinach. Biochem. Soc. Proc. 4, 77–80 (1976)

    Google Scholar 

  • Spencer, D., Wood, J.G.: The role of molybdenum in nitrate reductase in higher plants. Aust. J. biol. Sci. 7, 425–434 (1954)

    PubMed  Google Scholar 

  • Subramanian, K.N., Sorger, G.J.: The role of molybdenum in the synthesis of Neurospora nitrate reductase. Biochim. Biophys. Acta 256, 533–543 (1972)

    PubMed  Google Scholar 

  • Vega, J.M., Herrera, J., Aparicio, P.J., Paneque, A., Losada, M.: Role of molybdenum in nitrate reduction by Chlorella. Plant Physiol. 48, 294–299 (1971)

    Google Scholar 

  • Wallace, W.: Purification and properties of a nitrate reductase inactivating enzyme. Biochim. Biophys. Acta 341, 265–276 (1974)

    PubMed  Google Scholar 

  • Zielke, H.R., Filner, P.: Synthesis and turnover of nitrate reductase induced by nitrate in cultured tobacco cells. J. Biol. Chem. 246, 1772–1779 (1971)

    PubMed  Google Scholar 

Download references

Author information

Author notes

  1. R. W. Jones

    Present address: Dept. of Land Resource Science, University of Guelph, N1G 2W1, Ontario, Canada

Authors and Affiliations

  1. Long Ashton Research Station, University of Bristol, BS18 9AF, Bristol, UK

    R. W. Jones, A. J. Abbott, E. J. Hewitt, G. R. Best & E. F. Watson

Authors
  1. R. W. Jones
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. J. Abbott
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. E. J. Hewitt
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. G. R. Best
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. E. F. Watson
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jones, R.W., Abbott, A.J., Hewitt, E.J. et al. Nitrate reductase activity in Paul's scarlet rose suspension cultures and the differential role of nitrate and molybdenum in induction. Planta 141, 183–189 (1978). https://doi.org/10.1007/BF00387887

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00387887

Key words

Search

Navigation