Skip to main content
NCBI home page
The Plant Cell logoLink to The Plant Cell
. 1997 Nov;9(11):2077–2091. doi: 10.1105/tpc.9.11.2077

Early Events Induced by the Elicitor Cryptogein in Tobacco Cells: Involvement of a Plasma Membrane NADPH Oxidase and Activation of Glycolysis and the Pentose Phosphate Pathway.

A Pugin 1, J M Frachisse 1, E Tavernier 1, R Bligny 1, E Gout 1, R Douce 1, J Guern 1
PMCID: PMC157059  PMID: 12237354

Abstract

Application of the elicitor cryptogein to tobacco (cv Xanthi) is known to evoke external medium alkalinization, active oxygen species production, and phytoalexin synthesis. These are all dependent on an influx of calcium. We show here that cryptogein also induces calcium-dependent plasma membrane depolarization, chloride efflux, cytoplasm acidification, and NADPH oxidation without changes in NAD+ and ATP levels, indicating that the elicitor-activated redox system, responsible for active oxygen species production, uses NADPH in vivo. NADPH oxidation activates the functioning of the pentose phosphate pathway, leading to a decrease in glucose 6-phosphate and to the accumulation of glyceraldehyde 3-phosphate, 3- and 2-phosphoglyceric acid, and phosphoenolpyruvate. By inhibiting the pentose phosphate pathway, we demonstrate that the activation of the plasma membrane NADPH oxidase is responsible for active oxygen species production, external alkalinization, and acidification of the cytoplasm. A model is proposed for the organization of the cryptogein responses measured to date.

Full Text

The Full Text of this article is available as a PDF (1.3 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Apostol I., Heinstein P. F., Low P. S. Rapid Stimulation of an Oxidative Burst during Elicitation of Cultured Plant Cells : Role in Defense and Signal Transduction. Plant Physiol. 1989 May;90(1):109–116. doi: 10.1104/pp.90.1.109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Auh C. K., Murphy T. M. Plasma Membrane Redox Enzyme Is Involved in the Synthesis of O2- and H2O2 by Phytophthora Elicitor-Stimulated Rose Cells. Plant Physiol. 1995 Apr;107(4):1241–1247. doi: 10.1104/pp.107.4.1241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Babior B. M. The respiratory burst oxidase. Adv Enzymol Relat Areas Mol Biol. 1992;65:49–95. doi: 10.1002/9780470123119.ch2. [DOI] [PubMed] [Google Scholar]
  4. Basse C. W., Bock K., Boller T. Elicitors and suppressors of the defense response in tomato cells. Purification and characterization of glycopeptide elicitors and glycan suppressors generated by enzymatic cleavage of yeast invertase. J Biol Chem. 1992 May 25;267(15):10258–10265. [PubMed] [Google Scholar]
  5. Basse C. W., Fath A., Boller T. High affinity binding of a glycopeptide elicitor to tomato cells and microsomal membranes and displacement by specific glycan suppressors. J Biol Chem. 1993 Jul 15;268(20):14724–14731. [PubMed] [Google Scholar]
  6. Blein J. P., Milat M. L., Ricci P. Responses of Cultured Tobacco Cells to Cryptogein, a Proteinaceous Elicitor from Phytophthora cryptogea: Possible Plasmalemma Involvement. Plant Physiol. 1991 Feb;95(2):486–491. doi: 10.1104/pp.95.2.486. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bradley D. J., Kjellbom P., Lamb C. J. Elicitor- and wound-induced oxidative cross-linking of a proline-rich plant cell wall protein: a novel, rapid defense response. Cell. 1992 Jul 10;70(1):21–30. doi: 10.1016/0092-8674(92)90530-p. [DOI] [PubMed] [Google Scholar]
  8. Brisson L. F., Tenhaken R., Lamb C. Function of Oxidative Cross-Linking of Cell Wall Structural Proteins in Plant Disease Resistance. Plant Cell. 1994 Dec;6(12):1703–1712. doi: 10.1105/tpc.6.12.1703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Carpita N., McCann M., Griffing L. R. The plant extracellular matrix: news from the cell's frontier. Plant Cell. 1996 Sep;8(9):1451–1463. doi: 10.1105/tpc.8.9.1451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chanock S. J., el Benna J., Smith R. M., Babior B. M. The respiratory burst oxidase. J Biol Chem. 1994 Oct 7;269(40):24519–24522. [PubMed] [Google Scholar]
  11. Chen Z., Silva H., Klessig D. F. Active oxygen species in the induction of plant systemic acquired resistance by salicylic acid. Science. 1993 Dec 17;262(5141):1883–1886. doi: 10.1126/science.8266079. [DOI] [PubMed] [Google Scholar]
  12. Cross A. R., Jones O. T. The effect of the inhibitor diphenylene iodonium on the superoxide-generating system of neutrophils. Specific labelling of a component polypeptide of the oxidase. Biochem J. 1986 Jul 1;237(1):111–116. doi: 10.1042/bj2370111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Devlin W. S., Gustine D. L. Involvement of the oxidative burst in phytoalexin accumulation and the hypersensitive reaction. Plant Physiol. 1992 Nov;100(3):1189–1195. doi: 10.1104/pp.100.3.1189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Dwyer S. C., Legendre L., Low P. S., Leto T. L. Plant and human neutrophil oxidative burst complexes contain immunologically related proteins. Biochim Biophys Acta. 1996 Mar 15;1289(2):231–237. doi: 10.1016/0304-4165(95)00156-5. [DOI] [PubMed] [Google Scholar]
  15. Fahrendorf T., Ni W., Shorrosh B. S., Dixon R. A. Stress responses in alfalfa (Medicago sativa L.) XIX. Transcriptional activation of oxidative pentose phosphate pathway genes at the onset of the isoflavonoid phytoalexin response. Plant Mol Biol. 1995 Aug;28(5):885–900. doi: 10.1007/BF00042073. [DOI] [PubMed] [Google Scholar]
  16. GLASER L., BROWN D. H. Purification and properties of d-glucose-6-phosphate dehydrogenase. J Biol Chem. 1955 Sep;216(1):67–79. [PubMed] [Google Scholar]
  17. Gelli A., Higgins V. J., Blumwald E. Activation of Plant Plasma Membrane Ca2+-Permeable Channels by Race-Specific Fungal Elicitors. Plant Physiol. 1997 Jan;113(1):269–279. doi: 10.1104/pp.113.1.269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gilroy S., Read N. D., Trewavas A. J. Elevation of cytoplasmic calcium by caged calcium or caged inositol triphosphate initiates stomatal closure. Nature. 1990 Aug 23;346(6286):769–771. doi: 10.1038/346769a0. [DOI] [PubMed] [Google Scholar]
  19. Gout E., Bligny R., Douce R. Regulation of intracellular pH values in higher plant cells. Carbon-13 and phosphorus-31 nuclear magnetic resonance studies. J Biol Chem. 1992 Jul 15;267(20):13903–13909. [PubMed] [Google Scholar]
  20. Grabov A., Bottger M. Are Redox Reactions Involved in Regulation of K+ Channels in the Plasma Membrane of Limnobium stoloniferum Root Hairs? Plant Physiol. 1994 Jul;105(3):927–935. doi: 10.1104/pp.105.3.927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Greenberg J. T., Guo A., Klessig D. F., Ausubel F. M. Programmed cell death in plants: a pathogen-triggered response activated coordinately with multiple defense functions. Cell. 1994 May 20;77(4):551–563. doi: 10.1016/0092-8674(94)90217-8. [DOI] [PubMed] [Google Scholar]
  22. Greenspan H. C., Aruoma O. I. Oxidative stress and apoptosis in HIV infection: a role for plant-derived metabolites with synergistic antioxidant activity. Immunol Today. 1994 May;15(5):209–213. doi: 10.1016/0167-5699(94)90245-3. [DOI] [PubMed] [Google Scholar]
  23. Guern J., Mathieu Y., Kurkdjian A., Manigault P., Manigault J., Gillet B., Beloeil J. C., Lallemand J. Y. Regulation of Vacuolar pH of Plant Cells: II. A P NMR Study of the Modifications of Vacuolar pH in Isolated Vacuoles Induced by Proton Pumping and Cation/H Exchanges. Plant Physiol. 1989 Jan;89(1):27–36. doi: 10.1104/pp.89.1.27. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Isfort R. J., Cody D. B., Asquith T. N., Ridder G. M., Stuard S. B., LeBoeuf R. A. Induction of protein phosphorylation, protein synthesis, immediate-early-gene expression and cellular proliferation by intracellular pH modulation. Implications for the role of hydrogen ions in signal transduction. Eur J Biochem. 1993 Apr 1;213(1):349–357. doi: 10.1111/j.1432-1033.1993.tb17768.x. [DOI] [PubMed] [Google Scholar]
  25. Kieffer F., Simon-Plas F., Maume B. F., Blein J. P. Tobacco cells contain a protein, immunologically related to the neutrophil small G protein Rac2 and involved in elicitor-induced oxidative burst. FEBS Lett. 1997 Feb 17;403(2):149–153. doi: 10.1016/s0014-5793(97)00038-0. [DOI] [PubMed] [Google Scholar]
  26. Levine A., Tenhaken R., Dixon R., Lamb C. H2O2 from the oxidative burst orchestrates the plant hypersensitive disease resistance response. Cell. 1994 Nov 18;79(4):583–593. doi: 10.1016/0092-8674(94)90544-4. [DOI] [PubMed] [Google Scholar]
  27. Marrè M. T., Moroni A., Albergoni F. G., Marrè E. Plasmalemma redox activity and h extrusion: I. Activation of the h-pump by ferricyanide-induced potential depolarization and cytoplasm acidification. Plant Physiol. 1988 May;87(1):25–29. doi: 10.1104/pp.87.1.25. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Mehdy M. C. Active Oxygen Species in Plant Defense against Pathogens. Plant Physiol. 1994 Jun;105(2):467–472. doi: 10.1104/pp.105.2.467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Nishihara T., Akifusa S., Koseki T., Kato S., Muro M., Hanada N. Specific inhibitors of vacuolar type H(+)-ATPases induce apoptotic cell death. Biochem Biophys Res Commun. 1995 Jul 6;212(1):255–262. doi: 10.1006/bbrc.1995.1964. [DOI] [PubMed] [Google Scholar]
  30. Nürnberger T., Nennstiel D., Jabs T., Sacks W. R., Hahlbrock K., Scheel D. High affinity binding of a fungal oligopeptide elicitor to parsley plasma membranes triggers multiple defense responses. Cell. 1994 Aug 12;78(3):449–460. doi: 10.1016/0092-8674(94)90423-5. [DOI] [PubMed] [Google Scholar]
  31. Ojalvo I., Rokem J. S., Navon G., Goldberg I. P NMR Study of Elicitor Treated Phaseolus vulgaris Cell Suspension Cultures. Plant Physiol. 1987 Nov;85(3):716–719. doi: 10.1104/pp.85.3.716. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Petitot A. S., Blein J. P., Pugin A., Suty L. Cloning of two plant cDNAs encoding a beta-type proteasome subunit and a transformer-2-like SR-related protein: early induction of the corresponding genes in tobacco cells treated with cryptogein. Plant Mol Biol. 1997 Oct;35(3):261–269. doi: 10.1023/a:1005833216479. [DOI] [PubMed] [Google Scholar]
  33. Ricci P., Bonnet P., Huet J. C., Sallantin M., Beauvais-Cante F., Bruneteau M., Billard V., Michel G., Pernollet J. C. Structure and activity of proteins from pathogenic fungi Phytophthora eliciting necrosis and acquired resistance in tobacco. Eur J Biochem. 1989 Aug 15;183(3):555–563. doi: 10.1111/j.1432-1033.1989.tb21084.x. [DOI] [PubMed] [Google Scholar]
  34. Roberts J. K., Jardetzky O. Monitoring of cellular metabolism by NMR. Biochim Biophys Acta. 1981 Nov 9;639(1):53–76. doi: 10.1016/0304-4173(81)90005-7. [DOI] [PubMed] [Google Scholar]
  35. Roberts JKM., Aubert S., Gout E., Bligny R., Douce R. Cooperation and Competition between Adenylate Kinase, Nucleoside Diphosphokinase, Electron Transport, and ATP Synthase in Plant Mitochondria Studied by 31P-Nuclear Magnetic Resonance. Plant Physiol. 1997 Jan;113(1):191–199. doi: 10.1104/pp.113.1.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Roby C., Martin J. B., Bligny R., Douce R. Biochemical changes during sucrose deprivation in higher plant cells. Phosphorus-31 nuclear magnetic resonance studies. J Biol Chem. 1987 Apr 15;262(11):5000–5007. [PubMed] [Google Scholar]
  37. Rossi F. The O2- -forming NADPH oxidase of the phagocytes: nature, mechanisms of activation and function. Biochim Biophys Acta. 1986 Nov 4;853(1):65–89. doi: 10.1016/0304-4173(86)90005-4. [DOI] [PubMed] [Google Scholar]
  38. Rubinstein B., Stern A. I. Relationship of Transplasmalemma Redox Activity to Proton and Solute Transport by Roots of Zea mays. Plant Physiol. 1986 Apr;80(4):805–811. doi: 10.1104/pp.80.4.805. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Satre M., Martin J. B., Klein G. Methyl phosphonate as a 31P-NMR probe for intracellular pH measurements in Dictyostelium amoebae. Biochimie. 1989 Aug;71(8):941–948. doi: 10.1016/0300-9084(89)90076-x. [DOI] [PubMed] [Google Scholar]
  40. Schroeder J. I. Anion channels as central mechanisms for signal transduction in guard cells and putative functions in roots for plant-soil interactions. Plant Mol Biol. 1995 Jun;28(3):353–361. doi: 10.1007/BF00020385. [DOI] [PubMed] [Google Scholar]
  41. Schroeder J. I., Hedrich R. Involvement of ion channels and active transport in osmoregulation and signaling of higher plant cells. Trends Biochem Sci. 1989 May;14(5):187–192. doi: 10.1016/0968-0004(89)90272-7. [DOI] [PubMed] [Google Scholar]
  42. Segal A. W., Abo A. The biochemical basis of the NADPH oxidase of phagocytes. Trends Biochem Sci. 1993 Feb;18(2):43–47. doi: 10.1016/0968-0004(93)90051-n. [DOI] [PubMed] [Google Scholar]
  43. Segal A. W., Geisow M., Garcia R., Harper A., Miller R. The respiratory burst of phagocytic cells is associated with a rise in vacuolar pH. Nature. 1981 Apr 2;290(5805):406–409. doi: 10.1038/290406a0. [DOI] [PubMed] [Google Scholar]
  44. Slayman C. L., Long W. S., Lu C. Y. The relationship between ATP and an electrogenic pump in the plasma membrane of Neurospora crassa. J Membr Biol. 1973;14(4):305–338. doi: 10.1007/BF01868083. [DOI] [PubMed] [Google Scholar]
  45. Slonczewski J. L., Rosen B. P., Alger J. R., Macnab R. M. pH homeostasis in Escherichia coli: measurement by 31P nuclear magnetic resonance of methylphosphonate and phosphate. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6271–6275. doi: 10.1073/pnas.78.10.6271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Tavernier E., Wendehenne D., Blein J. P., Pugin A. Involvement of Free Calcium in Action of Cryptogein, a Proteinaceous Elicitor of Hypersensitive Reaction in Tobacco Cells. Plant Physiol. 1995 Nov;109(3):1025–1031. doi: 10.1104/pp.109.3.1025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Tenhaken R., Levine A., Brisson L. F., Dixon R. A., Lamb C. Function of the oxidative burst in hypersensitive disease resistance. Proc Natl Acad Sci U S A. 1995 May 9;92(10):4158–4163. doi: 10.1073/pnas.92.10.4158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Thuleau P., Ward J. M., Ranjeva R., Schroeder J. I. Voltage-dependent calcium-permeable channels in the plasma membrane of a higher plant cell. EMBO J. 1994 Jul 1;13(13):2970–2975. doi: 10.1002/j.1460-2075.1994.tb06595.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Viard M. P., Martin F., Pugin A., Ricci P., Blein J. P. Protein Phosphorylation Is Induced in Tobacco Cells by the Elicitor Cryptogein. Plant Physiol. 1994 Apr;104(4):1245–1249. doi: 10.1104/pp.104.4.1245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Ward J. M., Pei Z. M., Schroeder J. I. Roles of Ion Channels in Initiation of Signal Transduction in Higher Plants. Plant Cell. 1995 Jul;7(7):833–844. doi: 10.1105/tpc.7.7.833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Xing T., Higgins V. J., Blumwald E. Race-specific elicitors of Cladosporium fulvum promote translocation of cytosolic components of NADPH oxidase to the plasma membrane of tomato cells. Plant Cell. 1997 Feb;9(2):249–259. doi: 10.1105/tpc.9.2.249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Zimmermann S., Nürnberger T., Frachisse J. M., Wirtz W., Guern J., Hedrich R., Scheel D. Receptor-mediated activation of a plant Ca2+-permeable ion channel involved in pathogen defense. Proc Natl Acad Sci U S A. 1997 Mar 18;94(6):2751–2755. doi: 10.1073/pnas.94.6.2751. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Plant Cell are provided here courtesy of Oxford University Press

RESOURCES