Skip to main content
SpringerLink
Log in

Appearance of nitrite-reductase mRNA in mustard seedling cotyledons is regulated by phytochrome

  • Published:
Planta Aims and scope Submit manuscript

Abstract

Nitrate reductase (NR, EC 1.6.6.1) and nitrite reductase (NIR, EC 1.7.7.1) are the key enzymes of nitrate reduction. It is well established that the appearance of these enzymes is “induced” by nitrate, and it is generally believed that NR is cytosolic while NIR is plastidic. In mustard (Sinapis alba L.) cotyledons we observed two isoforms of NIR (NIR1 and NIR2) using a chromato-focusing technique. Only one of them (NIR2) disappeared when the plastids were damaged by photooxidation in the presence of Norflurazon. It is concluded that NIR2 is plastidic while NIR1 is extraplastidic and not affected by photooxidation of the plastids. Both isoforms appear to have the same molecular weight (60 kilodaltons, kDa). Two distinct translation products which could be immunoprecipitated with NIR antiserum produced against total NIR from mustard were observed which differed slightly in molecular weight (60 versus 63 kDa). The 63-kDa polypeptide was considered to be the precursor of NIR2. While synthesis of NIR protein depended largely on nitrate, the levels of in-vitro-translatable NIR mRNAs were found to be either independent of nitrate and light (NIR1) or controlled by phytochrome only (NIR2). It appears that phytochrome strongly stimulates the level of mRNA while significant enzyme synthesis (NIR2) takes place only in the presence of relatively large amounts of nitrate. Since an increased enzyme level was strictly correlated with an increase of immunoresponsive NIR protein it is improbable that activation of a precursor plays a role. Rather, it is concluded that, in situ, nitrate controls translation.

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

D:

darkness

FR:

far-red light (3.5 W m−2)

kDa:

kilodalton

Mr :

relative molecular mass

NF:

Norflurazon

NIR:

nitrite reductase (EC 1.7.7.1)

R:

strong red light(6.8 W. m−2)

SDS-PAGE:

sodium dodecyl sulphate-polyacryl-amide gel electrophoresis

References

  • Apel, K., Gollmer, I., Batschauer, A. (1983) The light-dependent control of chloroplast development in barley (Hordeum vulgare L.). J. Cell. Biochem. 23, 181–189

    Google Scholar 

  • Bonner, W.M., Laskey, R.A. (1974) A film detection method for tritium labelled proteins and nucleic acids in polyacrylamide gels. Eur. J. Biochem. 46, 83–88

    Google Scholar 

  • Brödenfeldt, R., Mohr, H. (1986) Use of immunotitration to demonstrate phytochrome-mediated synthesis de novo of chalcone synthase and phenylalanine ammonia lyase in mustard seedling cotyledons. Z. Naturforsch. 41c, 61–68

    Google Scholar 

  • Ellis, J.R. (1986) Photoregulation of plant gene expression. Biosci. Rep. 6, 127–136

    Google Scholar 

  • Frosch, S., Jabben, M., Bergfeld, R., Kleinig, H., Mohr, H. (1979) Inhibition of carotenoid biosynthesis by the herbicide SAN 9789and its consequences for the action of phytochrome on plastogenesis. Planta 145, 497–505

    Google Scholar 

  • Gallagher, T.F., Ellis, R.J. (1982) Light stimulated transcription of genes for two chloroplast polypeptides in isolated pea leaf nuclei. EMBO J. 1, 1493–1498

    Google Scholar 

  • Guerrero, M.G., Vega, J.M., Losada, M. (1981) The assimilatory nitrate-reducing system and its regulation. Annu. Rev. Plant Physiol. 32, 169–204

    Google Scholar 

  • Gupta, S.C., Beevers, L. (1985) Regulation of synthesis of nitrite reductase in pea leaves: In-vivo and in-vitro studies. Planta 166, 89–95

    Google Scholar 

  • Gupta, S.C., Beevers, L. (1987) Regulation of nitrite reductase. Cell-free translation and processing. Plant Physiol. 83, 750–754

    Google Scholar 

  • Ida, S., Mikami, B. (1986) Spinach ferredoxin-nitrite reductase: a purification procedure and characterization of chemical properties. Biochim. Biophys. Acta 871, 167–176

    Google Scholar 

  • Ida, S., Kobayakwa, K., Morita, Y. (1976) Ferredoxin-sepharose affinity chromatography for the purification of assimilatory nitrite reductase. FEBS Lett. 65, 305–308

    Google Scholar 

  • Kamin, H., Privalle, L.S. (1987) Nitrite reductase. In: Inorganic nitrogen metabolism, pp. 112–117, Ullrich, W.R., et al., eds. Springer, Berlin Heidelberg New York Tokyo

    Google Scholar 

  • Kaufman, L.S., Thompson, W.F., Briggs, W.R. (1984) Different red light requirements for phytochrome-induced accumulation of cab RNA and rbc S RNA. Science 226, 1447–1449

    Google Scholar 

  • Kramer, V., Lahners, K., Back, E., Privalle, L.S., Rothstein, S. (1989) Transient accumulation of nitrite reductase mRNA in maize following the addition of nitrate. Plant Physiol. 90, 1214–1220

    Google Scholar 

  • Kuhlemeier, C., Green, P.J., Chua, N.H. (1987) Regulation of gene expression in higher plants. Annu. Rev. Plant Physiol. 38, 221–257

    Google Scholar 

  • Kutscherra, M., Jost, W., Schlee, D. (1987) Isoenzymes of nitrite reductase in higher plants Occurrence, purification properties, and alteration during ontogenesis. J. Plant Physiol. 129, 383–393

    Google Scholar 

  • Lämmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685

    PubMed  Google Scholar 

  • Lamb, C.J., Lawton, M.A. (1983) Photocontrol of gene expression. In: Encyclopedia of plant physiology, N.S., vol. 16A: Photomorphogenesis, pp. 213–257, Shropshire, W., Mohr, H., eds. Springer, Berlin Heidelberg New York Tokyo

    Google Scholar 

  • Link, G. (1982) Phytochrome control of plastid mRNA in mustard (Sinapis alba L.) Planta 154, 81–86

    Google Scholar 

  • Mösinger, E., Batschauer, A., Schäfer, E., Apel, K. (1985) Phytochrome control of in vitro transcription of specific genes in isolated nuclei from barley (Hordeum vulgare). Eur. J. Biochem. 147, 137–142

    Google Scholar 

  • Mösinger, E., Batschauer, A., Vierstra, R., Apel, K., Schäfer, E. (1987) Comparisons of the effects of exogenous native phytochrome and in-vivo irradiation on in-vitro transcription in isolated nuclei from barley (Hordeum vulgare). Planta 170, 505–514

    Google Scholar 

  • Mohr, H. (1957) Der Einfluß monochromatischer Strahlung auf das Längenwachstum des Hypokotyls und auf die Anthocyanbildung bei Keimlingen von Sinapis alba L. Planta 49, 389–405

    Google Scholar 

  • Mohr, H. (1966) Untersuchungen zur phytochrominduzierten Phtomorphogenese des Senfkeimlings (Sinapis alba L.) Z. Pflanzenphysiol. 54, 63–68

    Google Scholar 

  • Mohr, H., Drumm-Herrel, H. (1981) Interaction between blue/UVlight and light operating through phytochrome in higher plants. In: Plants and the daylight spectrum, pp 423–441, Smith, H., ed. Academic Press, New York London

    Google Scholar 

  • Oelmüller, R., Mohr, H. (1986) Photooxidative destruction of cloroplasts and its consequences for expression of nuclear genes. Planta 167, 106–113

    Google Scholar 

  • Oelmüller, R., Schuster, C., Mohr, H. (1988) Physiological characterization of a plastidic signal required for nitrate-induced appearance of nitrate and nitrite reductases. Planta, 174, 75–83

    Google Scholar 

  • Rajasekhar, V.K., Mohr, H. (1986) Appearance of nitrite reductase in cotyledons of the mustard (Sinapis alba L.) seedling as affected by nitrate, phytochrome and photooxidative damage of plastids. Planta 168, 369–376

    Google Scholar 

  • Rajasekhar, V.K., Oelmüller, R. (1987) Regulation of induction of nitrate reductase and nitrite reductase in higher plants. Mini review. Physiol. Plant. 71, 517–521

    Google Scholar 

  • Reiss, T., Bergfeld, R., Link, G., Thien, W. and Mohr, H. (1983) Photooxidative destruction of chloroplasts and its consequences of cytosolic enzyme levels and plant development. Planta 159, 518–528

    Google Scholar 

  • Schmidt, S., Drumm-Herrel, H., Oelmüller, R., Mohr, H. (1987) Time course of competence in phytochrome-controlled appearance of nuclear encoded plastidic proteins and messenger RNAs. Planta 170, 400–407

    Google Scholar 

  • Schuster, C., Oelmüller, R., Mohr, H. (1987) Signal storage in phytochrome action on nitrate-mediated induction of nitrate reductase and nitrite reductase in mustard seedling cotyledons. Planta 171, 136–143

    Google Scholar 

  • Schuster, C., Oelmüller, R., Mohr, H. (1988a) Recovery of plastids from photooxidative damage: Significance of a plastidic factor. Planta 174, 289–297

    Google Scholar 

  • Schuster, C., Oelmüller, R., Mohr, H. (1988b) Control by phytochrome of the appearance of ribulose-1,5-bisphosphate carboxylase and the mRNA for its small subunit. Planta 174, 426–432

    Google Scholar 

  • Stiekema, W.J., Wimpee, C.F., Silverthorne, J., Tobin, E.M. (1983) Phytochrome control of the expression of two nuclear genes encoding chloroplast proteins in Lemna gibba L. Plant Physiol. 72, 717–724

    Google Scholar 

  • Suissa, M. (1983) Spectrophotometric quantitation of silver grains eluted from autoradiograms. Anal. Biochem. 133, 511–514

    Google Scholar 

  • Tobin, E.M., Silverthorne, J. (1985) Light regulation of gene expression in higher plants. Annu. Rev. Plant. Physiol. 36, 569–593

    Google Scholar 

  • Vega, J., Kamin, H. (1977) Spinach nitrite reductase: Purification and properties of a siroheme-containing iron-sulfur enzyme. J. Biol. Chem. 252, 896–909

    Google Scholar 

  • Weber, M., Schmidt, S., Schuster, C., Mohr, H. (1990) Factors involved in coordinate appearance of nitrite reductase and glutamine synthetase in the mustard (Sinapis alba L.) seedling. Planta 180, 429–434

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Biological Institute II, University of Freiburg, Schänzlestrasse 1, D-7800, Freiburg, Germany

    C. Schuster & H. Mohr

Authors
  1. C. Schuster
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. H. Mohr
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

This research was supported by a grant from the Deutsche Forschungsgemeinschaft. We thank I. Mehrer and A. Rashid for their contributions to Fig. 8. We are indebted to Mrs E. Janssen for writing the manuscript and to Mrs U. Meurer and Mrs G. Amschlinger for preparing the figures.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schuster, C., Mohr, H. Appearance of nitrite-reductase mRNA in mustard seedling cotyledons is regulated by phytochrome. Planta 181, 327–334 (1990). https://doi.org/10.1007/BF00195884

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00195884

Key words

Search

Navigation