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Evidence for the nitrate-dependent spatial regulation of the nitrate reductase gene in chicory roots

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Abstract

Young chicory plants (Cichorium intybus L. var. Witloof) show a tenfold higher nitrate reductase NR activity in roots compared to leaves. Northern analysis revealed, besides the nitrate inducibility of the nitrate reductase gene (nia), a higher level of expression in the roots. By modifying the external nitrate concentration the NR activity in the leaves remained negligible whereas a maximal activity was observed in the roots when grown in the presence of 5 mM nitrate. Surprisingly, variation of the external nitrate concentration induced changes in the spatial regulation of nia within the root. In-situ hybridization mainly localized nia mRNA in the cortical cells of roots grown at low nitrate concentrations (0.2 mM). At high nitrate concentrations (5 mM), nia mRNA was more abundant in the vascular tissues. The root apex revealed a strong signal under both conditions. The isolation and characterization of the NR structural gene from chicory is also presented. Southern blot analysis revealed the presence of a single nia gene per haploid genome of chicory.

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Abbreviations

cNRs:

chicory nitrate reductase cDNA

DIG:

digoxigenin

MoCo:

molybdenum cofactor

NiR:

nitrite reductase

NG:

nitrate reductase genomic sequence

NR:

nitrate reductase

NRA:

nitrate reductase activity

PCR:

polymerase chain reaction

References

  • Andrews M (1986) The partitioning of nitrate assimilation between root and shoot of higher plants. Plant Cell Environ 9: 511–519

    Google Scholar 

  • Cheng CL, Acedo GN, Cristinsin M, Conkling MA (1992) Sucrose mimics the light induction of Arabidopsis nitrate reductase gene transcription. Proc Natl Acad Sci USA 89: 1861–1864

    Google Scholar 

  • Choi-Hong K, Kleinhofs A, An G (1989) Nucleotide sequence of rice nitrate reductase genes. Plant Mol Biol 13: 731–733

    Google Scholar 

  • Church G, Gilbert W (1984) Genomic sequencing. Proc Natl Acad Sci USA 81: 1991–1995

    CAS  PubMed  Google Scholar 

  • Collins FS, Weissman SM (1984) Directional cloning of DNA fragments at a large distance from an intial probe: A circularization method. Proc Natl Acad Sci USA 81: 6812–6816

    Google Scholar 

  • Cox KH, De Leon DV, Angerer LM, Angerer RC (1984) Detection of mRNAs in sea urchin embryos by in situ hybridization using assymetric RNA probes. Dev Biol 101: 485–502

    Google Scholar 

  • Crawford NM, Arst Jr HN (1993) The molecular genetics of nitrate assimilation in fungi and plants. Annu Rev Genet 27: 115–146

    Google Scholar 

  • Crawford NM, Campbell WH, Davis RW (1986) Nitrate reductase from squash: cDNA cloning and nitrate regulation. Proc Natl Acad Sci USA 83: 8073–8076

    Google Scholar 

  • Cyr DR, Bewley JD, Dumbroff EB (1990) Seasonal dynamics of carbohydrates and nitrogenous compounds in the roots of perennial weeds. Plant Cell Environ 13: 359–365

    Google Scholar 

  • Daniel-Vedele F, Dorbe MF, Caboche M, Rouze P (1989) Cloning and analysis of the nitrate reductase gene from tomato: a comparison of nitrate reductase protein sequences in higher plants. Gene 85: 371–380

    Google Scholar 

  • de Almeida Engler J, Van Montagu M, Engler G (1994) Whole mount in situ mRNA hybridization in plants. Plant Mol Biol Rep 12: 321–331

    Google Scholar 

  • Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: Version II. Plant Mol Biol Rep 1: 19–21

    CAS  Google Scholar 

  • Deng MD, Faure JD, Caboche M (1993) The molecular aspects of nitrate reductase expression in higher plants. In: Verma DPS (ed) Control of gene expression. Springer, Wien-New York, pp 425–441

    Google Scholar 

  • Dorchies V, Rambour S (1985) Activité nitrate reductase chez Cichorium intybus (var.Witloof) à différents stades de développement et dans les tissus cultivés in vitro. Physiol Vég 23: 25–35

    Google Scholar 

  • Faure JD, Vincentz M, Kronenberger J, Caboche M (1991) Coregulated expression of nitrate and nitrite reductases. Plant J 1: 107–113

    Google Scholar 

  • Federova E, Greenwood JS, Oaks A (1994) In situ localization of nitrate reductase in maize roots. Planta 194: 279–286

    Google Scholar 

  • Gupta AK, Mann P, Kaur N, Singh R (1991) Profiles of enzymes of sucrose metabolism in the leaves of chicory (Cichorium intybus) during development. Plant Sci 77: 191–196

    Google Scholar 

  • Hoff T, Stummann BM, Henningsen KW (1991) Cloning and expression of a gene encoding a root specific nitrate reductase in bean (Phaseolus vulgari). Physiol Plant 82: 197–204

    Google Scholar 

  • Hoff T, Truong H-N, Caboche M (1994) The use of mutants and transgenic plants to study nitrate assimilation. Plant Cell Environ 17: 489–506

    Google Scholar 

  • Huppe HC, Turpin DH (1994) Integration of carbon and nitrogen metabolism in plant and algae cells. Annu Rev Plant Physiol Plant Mol Biol 45: 577–607

    Google Scholar 

  • Huppe HC, Farr TJ, Turpin DH (1994) Coordination of chloroplastic metabolism in N-limited Chlamydomonas reinhardtii by redox modulation. Plant Physiol 105: 1043–1048

    Google Scholar 

  • Jaworski EG (1971) Nitrate reductase assay in intact plant tissues. Biochem Biophys Res Commun 43: 1274–1279

    Google Scholar 

  • Kaiser WM, Huber SC (1994) Posttranslational regulation of nitrate reductase in higher plants. Plant Physiol 106: 817–821

    Google Scholar 

  • Kronenberger J, Desprez T, Höfte H, Caboche M, Traas J (1993) A methacrylate embedding procedure developed for immunolocalization on plant tissues is also compatible with in situ hybridization. Cell Biol Int 17: 1013–1021

    Google Scholar 

  • Lillo C (1994) Light regulation of nitrate reductase in green leaves of higher plants. Physiol Plant 90: 616–620

    Google Scholar 

  • Limami A, Roux L, Laville J, Roux Y (1993) Dynamics of nitrogen compounds in the chicory (Cichorium intybus L.) tuberised tap root during the growing season and cold storage period. J Plant Physiol 141: 263–268

    Google Scholar 

  • Logemann J, Schell J, Willmitzer L (1987) Improved method for the isolation of RNA from plant tissues. Anal Biochem 163: 16–20

    CAS  PubMed  Google Scholar 

  • Long DM, Oaks A (1990) Stabilization of nitrate reductase in maize roots by chymostatin. Plant Physiol 93: 846–850

    Google Scholar 

  • Long DM, Oaks A, Rothstein SJ (1992) Regulation of maize root nitrate reductase mRNA levels. Physiol Plant 85: 561–566

    Google Scholar 

  • Miyazaki J, Juricek M, Angelis K, Schnorr KM, Kleinhofs A, Warner RL (1991) Characterization and sequence of a novel nitrate reductase from barley. Mol Gen Genet 228: 329–334

    Google Scholar 

  • Murashige T, Skoog K (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497

    CAS  Google Scholar 

  • Oaks A (1994a) Primary nitrogen assimilation in higher plants and its regulation. Can J Bot: 739–750

  • Oaks A (1994b) Efficiency of nitrogen utilization in C3 and C4 cereals. Plant Physiol 106: 407–414

    Google Scholar 

  • Presser IM, Lazarus CM (1990) Nucleotide sequence of a spinach nitrate reductase cDNA. Plant Mol Biol 15: 187–190

    Google Scholar 

  • Rastogi R, Back E, Schneiderbauer A, Bowscher CG, Moffatt B, Rothstein SJ (1993) A 330 bp region of the spinach nitrite reductase gene promoter directs nitrate-inducible tissue-specific expression in transgenic tobacco. Plant J 4: 317–326

    Google Scholar 

  • Rufty TW, Thomas JF, Remmler J, Campbell WH, Volk RJ (1986) Intracellular localization of nitrate reductase in roots. Plant Physiol 82: 675–680

    Google Scholar 

  • Saiki RK, Gelfland DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA (1988) Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science: 487–491

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning, a laboratory manual. 2nd edn. Cold-Spring Harbor Laboratory, New York

    Google Scholar 

  • Sechley KA, Oaks A, Bewley JD (1991) Enzymes of nitrogen assimilation undergo seasonal fluctuations in the roots of the persistent weedy perennial Cichorium intybus. Plant Physiol 97: 322–329

    Google Scholar 

  • Shaner DC, Boyer JS (1976) Nitrate reductase activity in maize (Zea mays L.) leaves. I. Regulation by nitrate flux. Plant Physiol 58: 499–504

    Google Scholar 

  • Vaucheret H, Vincentz M, Kronenberger J, Caboche M, Rouze P (1989) Molecular cloning and characterization of the two homologous genes coding for nitrate reductase in tobacco. Mol Gen Genet 216: 10–15

    Google Scholar 

  • Verwoerd TC, Dekker BMM, Hoekema A (1989) A small-scale procedure for the rapid isolation of plant RNAs. Nucleic Acids Res 6: 2362

    Google Scholar 

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Authors and Affiliations

  1. Laboratoire de Physiologie et Génétique Moléculaire Végétales, Université des Sciences et Technologies de Lille, Bâtiment SN2, F-59655, Villeneuve d'Ascq Cedex, France

    Benoît Palms, Pascale Goupil & Serge Rambour

  2. Laboratorium voor Genetica, Ledeganckstraat 35, B-9000, Gent, Belgium

    Janice de Almeida Engler, Dominique Van Der Straeten & Marc Van Montagu

Authors
  1. Benoît Palms
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  2. Pascale Goupil
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  3. Janice de Almeida Engler
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  4. Dominique Van Der Straeten
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  5. Marc Van Montagu
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  6. Serge Rambour
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Corresponding author

Correspondence to Serge Rambour.

Additional information

The authors wish to thank W. Ardiles Diaz, J. Gielen and R. Villarroel from the Lab. of Genetics, Gent for the DNA sequencing. We are indebted to P. Rouze and G. Engler for critical reading of the manuscript. B.P. is indebted to the European Molecular Biology Organization (Heidelberg, Germany) for a short-term fellowship. D.V.D.S. is a Senior Research Associate of the National Fund for Scientific Research (Belgium). This work was supported by the Conseil Regional Nord-Pas-de-Calais, France.

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Palms, B., Goupil, P., de Almeida Engler, J. et al. Evidence for the nitrate-dependent spatial regulation of the nitrate reductase gene in chicory roots. Planta 200, 20–27 (1996). https://doi.org/10.1007/BF00196644

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  • DOI: https://doi.org/10.1007/BF00196644

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