, Volume 197, Issue 4, pp 613–618 | Cite as

Evidence for the involvement of plasma-membrane-bound nitrate reductase in signal transduction during blue-light stimulation of nitrate uptake in Chlorella saccharophila

  • Christine Stöhr
  • Ulrich Glogau
  • Michael Mätschke
  • Rudolf Tischner


Nitrate uptake in Chlorella saccharophila (Krüger) Nadson was found to be stimulated by blue light, leading to a doubling of the rate. In the presence of background red light (300 μmol photons · m-2 · s-1), only 15–20 μmol photons · m-2 · s-1 of blue light was sufficient to saturate this increased uptake rate. Incubation of Chlorella cells with anti-nitrate-reductase immunoglobulin-G fragments inhibited blue-light stimulation. However, ferricyanide (10 μM) doubled and dithiothreitol (100 μM) inhibited the stimulatory effect of blue light. Among the protein-kinase inhibitors used, only staurosporine (10 μM) prevented the blue-light stimulation. Phosphatase inhibitors were without effect and sodium vanadate totally inhibited nitrate uptake, pointing to an involvement of the plasma-membrane ATPase. Preincubation of the cells with calmodulin antagonists or calcium ionophores did not significantly reduce blue-light stimulation of nitrate uptake. The data are discussed with regard to transduction of the signal for blue-light stimulation of nitrate uptake and the possibility that the plasma-membrane-bound nitrate reductase is the blue-light receptor.

Key words

Blue-light Chlorella Nitrate reductase (plasma-membrane-bound) Nitrate uptake 











nitrate reductase




immunoglobulin G


photon flux density


plasma membrane




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  1. Achmad M, Cashmore AR (1993) HY4 gene of Arabidopsis thaliana encodes a protein with characteristics of a blue-light photo-receptor. Nature 366: 162–166Google Scholar
  2. Amodeo G, Srivastava A, Zeiger E (1992) Vanadate inhibits blue light-stimulated swelling of Vicia guard cell protoplasts. Plant Physiol 100: 1567–1570Google Scholar
  3. Aparichio PJ, Quinones MA (1991) Blue light, a positive switch signal for nitrate and nitrite uptake by the green alga Monoraphidium braunii. Plant Physiol 95: 374–378Google Scholar
  4. Aparichio PJ, Roldan JM, Calero F (1976) Blue light photoreactivation of nitrate reductase from green algae and higher plants. Biochem Biophys Res Commun 70: 1071–1077Google Scholar
  5. Askerlund P, Larsson C, Widell S (1989) Cytochromes of plant membranes.Characterization by absorbance difference spectroscopy and redox titration. Physiol Plant 76: 123–134Google Scholar
  6. Assmann SM, Simoncini L, Schroeder JI (1985) Blue light activates electronic ion pumping in guard cell protoplasts of Vicia faba. Nature 318: 285–286Google Scholar
  7. Brain RD, Freeburg JA, Weiss CV, Briggs WR (1977) Blue light-induced absorbance changes in membrane fractions from corn and Neurospora. Plant Physiol 59: 948–952Google Scholar
  8. Briggs WR, Iino M (1983) Blue light absorbing photoreceptors in plants. Phil Trans R Soc London Ser B 303: 347–359Google Scholar
  9. Calero F, Ullrich WR, Aparichio PJ (1980) Regulation by monochromatic light of nitrate uptake in Chlorella fusca. In: Senger H (ed) The blue light syndrome. Springer, Berlin, pp 411–421Google Scholar
  10. Cohen P, Holmes CFB, Tsukitani Y (1990) Okadaic acid: a new probe for the study of cellular regulation. Trends Biochem Sci 15: 98–102Google Scholar
  11. Corzo A, Plasa R, Ullrich WR (1991) Extracellular ferricyanide reduction and nitrate reductase activity in the green alga Monoraphidium braunii. Plant Sci 79: 221–228Google Scholar
  12. De la Rosa MA, Roncel M, Navarro JA (1989) Flavin-mediated photoregulation of nitrate reductase.A key point of control in inorganic nitrogen photosynthetic metabolism. Bioelectrochem Bioenerg 22: 355–364Google Scholar
  13. Dharmawardhane S, Rubinstein B, Stern AI (1989) Regulation of transplasmalemma electron transport in oat mesophyll cells by sphingoid bases and the blue light. Plant Physiol 89: 1345–1350Google Scholar
  14. Einspahr KJ, Thompson GA (1990) Transmembrane signalling via phosphatidylinositol 4,5-biphosphate hydrolysis in plants. Plant Physiol 93: 361–366Google Scholar
  15. Gallagher S, Leonard RT (1982) Electrophoretic characterization of a detergent-treated plasma membrane fraction from corn roots. Plant Physiol 83: 265–271Google Scholar
  16. Galland P, Senger H (1988) The role of pterins in the photoreception and metabolism of plants. Photochem Photobiol 48: 811–820Google Scholar
  17. Gautier H, Vavasseur A, Lasceve G, Boudet AM (1992) Redox processes in the blue light responses of guard cell protoplasts of Commelina communis L. Plant Physiol 98: 34–38Google Scholar
  18. Hager A, Brich M (1993) Blue-light-induced phosphorylation of a plasma membrane protein from phototropically sensitive tips of maize coleoptiles. Planta 189: 567–576Google Scholar
  19. Hannum YA, Bell RM (1989) Function of sphingolipids and sphingolipid breakdown products in cellular regulation. Science 243: 500–506Google Scholar
  20. Herbert JM, Seban E, Maffrand JP (1990) Characterization of specific binding sites for [3H]-staurosporine in various protein kinases. Biochem Biophys Res Commun 171: 189–195Google Scholar
  21. Huber JLA, Huber SC, Hamborg NT (1989) Protein phosphorylation as a mechanism for regulation of spinach leaf sucrose-phosphate synthase activity. Arch Biochem Biophys 270: 681–690Google Scholar
  22. Jetschmann K, Solomonson LP, Vennesland B (1972) Activation of nitrate reductase by oxidation. Biochim Biophys Acta 275: 276–278Google Scholar
  23. Lee Y, Assmann SM (1991) Diacylglyerols induce both ion pumping in patch-clamped guard-cell protoplasts and opening of intact stomata. Proc Natl Acad Sci USA 88: 2127–2131Google Scholar
  24. Lifter J, Choi YS (1978) Separation of IgG Fab and Fc fragments by isoelectric focussing. J Immun Methods 23: 297–302Google Scholar
  25. Luster DG, Buckhout TJ (1989) Purification and identification of a plasma membrane associated electron transport protein from maize (Zea mays L.) roots. Plant Physiol 91: 1014–1019Google Scholar
  26. McCurdy DW, Harmon AC (1992) Calcium-dependent protein kinase in the green alga Chara. Planta 188: 54–61Google Scholar
  27. McNaughton GAL, Mckintosh C, Fewson CA, Wilkins MB, Nimmo HG (1991) Illumination increases the phosphorylation state of maize leaf phosphoenolpyruvate carboxylase by causing an increase in the activity of a protein kinase. Biochim Biophys Acta 1093: 189–195Google Scholar
  28. Ninnemann H, Klemm-Wolfgramm E (1980) Blue light-controlled conidiation and absorbance change in Neurospora are mediated by nitrate reductase. In: Senger H (ed) The blue light syndrome Springer, Berlin, pp 238–243Google Scholar
  29. Ranjeva R, Boudet A (1987) Phosphorylation of proteins in plants: Regulatory effects and potential involvement in stimulus/ response coupling. Annu Rev Plant Physiol 38: 73–93Google Scholar
  30. Rubinstein B, Luster DG (1993) Plasma membrane redox activity: Components and role in plant processes. Annu Rev Plant Physiol 44: 131–155Google Scholar
  31. Roldan JM, Calero F, Aparichio PJ (1978) Photoreactivation of spinach nitrate reductase: role of flavins. Z Pflanzenphysiol 82: 261–273Google Scholar
  32. Shimazaki K, Kinoshita T, Nishimura M (1992) Involvement of calmodulin and calmodulin-dependent myosin light chain kinase in blue light-dependent H+ pumping by guard cell protoplasts from Vicia faba L. Plant Physiol 99: 1416–1421Google Scholar
  33. Short TW, Briggs WR (1990) Characterization of a rapid, blue light-mediated change in detectable phosphorylation of a plasma membrane protein from etiolated pea (Pisum sativum L.) seedlings. Plant Physiol 92: 179–185Google Scholar
  34. Short TW, Reymond P, Briggs WR (1993) A pea plasma membrane protein exhibiting blue light-induced phosphorylation retains photosensitivity following Triton solubilization. Plant Physiol 101: 647–655Google Scholar
  35. Stöhr Ch, Tischner R, Ward MR (1993) Characterization of the plasma-membrane-bound nitrate reductase in Chlorella saccharophila (Krüger) Nadson. Planta 191: 79–85Google Scholar
  36. Stöhr Ch, Schuler F, Tischner R (1995) Glycosyl-phosphatidyl-inositol anchored proteins exist in the plasma membrane of Chlorella saccharophila (Krüger) Nadson: Plasma-membrane-bound nitrate reductase as an example. Planta 196: 284–287Google Scholar
  37. Solomonson LP, Barber MJ, Robbins AP, Oaks A (1986) Functional domains of assimilatory NADH: nitrate reductase of Chlorella. J Biol Chem 261: 11290–11294Google Scholar
  38. Thayer JR, Huffaker RC (1980) Determination of nitrate and nitrite by high-pressure liquid chromatography: Comparison with other methods for nitrate determination. Analyt Biochem 102:110–119Google Scholar
  39. Tischner R (1984) A comparison of the high-active and low-active form of nitrate reductase in synchronous Chlorella sorokiniana. Planta 139: 1–5Google Scholar
  40. Tischner R, Lorenzen H (1979) Nitrate uptake and nitrate reduction in synchronous Chlorella. Planta 146: 287–292Google Scholar
  41. Tischner R, Hillmer S, Robinson DG (1987) Physiological properties and cytological features of protoplasts prepared from Chlorella saccharophila. Protoplasma 139: 153–159Google Scholar
  42. Tischner R, Ward MR, Huffaker RC (1989) Evidence for a plasma-membrane-bound nitrate reductase involved in nitrate uptake of Chlorella sorokiniana. Planta 178: 19–24Google Scholar
  43. Ullrich WR (1987) Nitrate and ammonium uptake in green algae and higher plants: mechanism and relationship with nitrate metabolism. In: Ullrich WR (ed) Inorganic nitrogen metabolism, Springer, Berlin, pp 32–38Google Scholar
  44. Ullrich WR, Novacky A (1981) Nitrate-dependent membrane potential changes and their induction in Lemna gibba G1. Plant Sci Lett 22: 211–217Google Scholar
  45. Ward MR, Tischner R, Huffaker RC (1988) Inhibition of nitrate transport by anti-nitrate reductase IgG fragments and the identification of plasma membrane associated nitrate reductase in roots of barley seedlings. Plant Physiol 88: 1141–1145Google Scholar

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • Christine Stöhr
    • 1
  • Ulrich Glogau
    • 2
  • Michael Mätschke
    • 2
  • Rudolf Tischner
    • 2
  1. 1.Institut für Botanik, Technische HochschuleDarmstadtGermany
  2. 2.Pflanzenphysiologisches Institut der Universität GöttingenGöttingenGermany

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