Skip to main content
Log in

Nitrite reduction in barley-root plastids: Dependence on NADPH coupled with glucose-6-phosphate and 6-phosphogluconate dehydrogenases, and possible involvement of an electron carrier and a diaphorase

  • Published:
Planta Aims and scope Submit manuscript

Abstract

Plastids from roots of barley (Hordeum vulgare L.) seedlings were isolated by discontinuous Percoll-gradient centrifugation. Coinciding with the peak of nitrite reductase (NiR; EC 1.7.7.1, a marker enzyme for plastids) in the gradients was a peak of a glucose-6-phosphate (Glc6P) and NADP+-linked nitrite-reductase system. High activities of phosphohexose isomerase (EC 5.3.1.9) and phosphoglucomutase (EC 2.7.5.1) as well as glucose-6-phosphate dehydrogenase (Glc6PDH; EC 1.1.1.49) and 6-phosphogluconate dehydrogenase (6PGDH; EC 1.1.1.44) were also present in the isolated plastids. Thus, the plastids contained an overall electron-transport system from NADPH coupled with Glc6PDH and 6PGDH to nitrite, from which ammonium is formed stoichiometrically. However, NADPH alone did not serve as an electron donor for nitrite reduction, although NADPH with Glc6P added was effective. Benzyl and methyl viologens were enzymatically reduced by plastid extract in the presence of Glc6P+ NADP+. When the plastids were incubated with dithionite, nitrite reduction took place, and ammonium was formed stoichiometrically. The results indicate that both an electron carrier and a diaphorase having ferredoxin-NADP+ reductase activity are involved in the electron-transport system of root plastids from NADPH, coupled with Glc6PDH and 6PGDH, to nitrite.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

Cyt:

cytochrome

Glc6P:

glucose-6-phosphate

Glc6PDH:

glucose-6-phosphate dehydrogenase

MVH:

reduced methyl viologen

NiR:

nitrite reductase

6PG:

6-phosphogluconate

6PGDH:

6-phosphogluconate dehydrogenase

References

  • Abrol, Y.P., Sawhner, S.K., Naik, M.S. (1983) Light and dark assimilation of nitrate in plants. Plant Cell Environ. 6, 595–599

    Google Scholar 

  • Aebi, H. (1974) Catalase. In: Methods of enzymatic analysis, vol. 2, pp. 673–684, Bergmeyer, H.U., ed. Academic Press, New York London

    Google Scholar 

  • Dalling, M.J., Tolbert, N.E., Hageman, R.H. (1972) Intracellular location of nitrate reductase and nitrite reductase. II. Wheat roots. Biochim. Biophys. Acta 283, 513–515

    Google Scholar 

  • Dalling, M.J., Hucklesby, D.P., Hageman, R.H. (1973) A comparison of nitrite reductase enzymes from green leaves, scutella, and roots of corn (Zea mays L.). Plant Physiol. 51, 481–484

    Google Scholar 

  • Dulley, J.R., Grieve, P.A. (1975) A simple technique for eliminating interference by detergents in the Lowry method of protein determination. Anal. Biochem. 64, 136–141

    Google Scholar 

  • Emes, M.J., Fowler, M.W. (1979) Intracellular interactions between the pathways of carbohydrate oxidation and nitrate assimilation in plant roots. Planta 145, 287–292

    Google Scholar 

  • Emes, M.J., Fowler, M.W. (1983) The supply of reducing power for nitrite reduction in plastids of seedlings pea root (Pisum sativum L.). Planta 158, 97–102

    Google Scholar 

  • Hackett, D.P. (1964) Enzymes of terminal respiration. In: Modern methods of plant analysis, vol. 7, pp. 647–694, Linskens, H.F., Sanwal, B.W., Tracey, M.V., eds. Springer, Berlin Göttingen Heidelberg

    Google Scholar 

  • Hucklesby, D.P., Dalling, M.J., Hageman, R.H. (1972) Some properties of two forms of nitrite reductase from corn (Zea mays L.) scutellum. Planta 104, 220–233

    Google Scholar 

  • Ida, S., Mori, E., Morita, Y. (1974) Purification, stabilization and characterization of nitrite reductase from barley roots. Planta 121, 213–224

    Google Scholar 

  • Joy, K.W., Hageman, R.H. (1966) The purification and properties of nitrite reductase from higher plants, and its dependence on ferredoxin. Biochem. J. 100, 263–273

    Google Scholar 

  • Kaplan, A. (1969) The determination of urea, ammonia, and urease. Methods Biochem. Anal 17, 311–324

    Google Scholar 

  • Lee, R.B. (1980) Sources of reductant for nitrate assimilation in non-photosynthetic tissue: a review. Plant Cell Environ. 3, 65–90

    Google Scholar 

  • Matoh, T., Takahashi, E. (1982) Changes in the activities of ferredoxin- and NADH-glutamate synthase during seedling development of pea. Planta 154, 289–294

    Google Scholar 

  • Miflin, B.J. (1974) The location of nitrite reductase and other enzymes related to amino acid biosynthesis in the plastids of root and leaves. Plant Physiol. 54, 550–555

    Google Scholar 

  • Ninomiya, Y., Sato, S. (1983) An electron carrier in nitrite reduction in proplastids of cultured tobacco cells. Plant Cell Tissue Organ Cult. 2, 285–292

    Google Scholar 

  • Ninomiya, Y., Sato, S. (1984) A ferredoxin-like electron carrier from non-green cultured tobacco cells. Plant Cell Physiol. 25, 453–458

    Google Scholar 

  • Oji, Y., Izawa, G. (1969) Effects of univalent cations on the formation of nitrate reductase and nitrite reductase in rice seedlings. Plant Cell Physiol. 10, 665–674

    Google Scholar 

  • Oji, Y., Miki, Y., Okamoto, S. (1982) Extraction and affinity purification of NADH: nitrate reductase from barley (Hordeum distichum L.) root. Plant Cell Physiol. 23, 1025–1031

    Google Scholar 

  • Paneque, A., Ramirez, J.M., Del Campo, E.F., Losada, M. (1964) Light and dark reduction of nitrite in a reconstituted enzymic system. J. Biol. Chem. 239, 1737–1741

    Google Scholar 

  • Sanderson, G.W., Cocking, E.C. (1964) Enzymic assimilation of nitrate in tomato plants. II. Reduction of nitrite to ammonia. Plant Physiol. 39, 423–431

    Google Scholar 

  • Scholl, R.L., Harper, J.E., Hageman, R.H. (1974) Improvements of the nitrite color development in assays of nitrate reductase by phenazine methosulfate and zinc acetate. Plant Physiol. 53, 825–828

    Google Scholar 

  • Senn, D.R., Carr, P.W., Klatt, L.N. (1976) Minimization of a sodium dithionite-derived interference in nitrate reductase-methyl viologen reagents. Anal. Biochem. 75, 464–471

    Google Scholar 

  • Suzuki, A., Vidal, J., Gadal, P. (1982) Glutamate synthase isoforms in rice. Immunological studies of enzymes in green leaf, etiolated leaf and root tissues. Plant Physiol. 70, 827–832

    Google Scholar 

  • Washitani, I., Sato, S. (1977a). Studies on the function of proplastids in the metabolism of in vitro-cultured tobacco cells. I. Localization of nitrite reductase and NADP-dependent glutamate dehydrogenase. Plant Cell Physiol. 18, 117–125

    Google Scholar 

  • Washitani, I., Sato, S. (1977b) Studies on the function of proplastids in the metabolism of in vitro-cultured tobacco cells. III. Source of reducing power for amino acid synthesis from nitrite. Plant Cell Physiol. 18, 1235–1241

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Oji, Y., Watanabe, M., Wakiuchi, N. et al. Nitrite reduction in barley-root plastids: Dependence on NADPH coupled with glucose-6-phosphate and 6-phosphogluconate dehydrogenases, and possible involvement of an electron carrier and a diaphorase. Planta 165, 85–90 (1985). https://doi.org/10.1007/BF00392215

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00392215

Key words

Navigation