Abstract
Stressed plant cells often show increased oxygen uptake which can manifest itself in the transient production of active oxygen species, the oxidative burst. There is a lack of information on the redox status of cells during the early stages of biotic stress. In this paper we measure oxygen uptake and the levels of redox intermediates NAD/NADH and ATP and show the transient induction of the marker enzyme for redox stress, alcohol dehydrogenase. Rapid changes in the redox potential of elicitor-treated suspension cultures of French bean cells indicate that, paradoxically, during the period of maximum oxygen uptake the levels of ATP and the NADH/NAD ratio fall in a way that indicates the occurrence of stress in oxidative metabolism. This period coincides with the maximum production of active oxygen species particularly H2O2. The cells recover and start producing ATP immediately upon the cessation of H2O2 production. This indicates that the increased O2 uptake is primarily incorporated into active O2 species. A second consequence of these changes is probably a transient compromising of the respiratory status of the cells as indicated in expression of alcohol dehydrogenase. Elicitor-induced bean ADH was purified to homogeneity and the Mr 40 000 polypeptide was subjected to amino acid sequencing. 15% of the whole protein was sequenced from three peptides and was found to have nearly 100% sequence similarity to the amino acid sequence for pea ADH1 (PSADH1). The cDNA coding for the pea enzyme was used to demonstrate the transient induction of ADH mRNA in elicitor-treated bean cells. Enzyme activity levels also increased transiently subsequently. Increased oxygen uptake has previously been thought to be associated with provision of energy for the changes in biosynthesis that occur rapidly after perception of the stress signal. However the present work shows that this rapid increase in oxygen uptake as a consequence of elicitor action is not wholly associated with respiration.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Andrews DL, Cobb BG, Johnson JR, Drew MC: Hypoxic and anoxic induction of alcohol dehydrogenase in roots and shoots of seedlings of Zea mays. Plant Physiol 101: 407–414 (1993).
Baker CJ, Orlandi EW, Mock NM: Harpin, an elicitor of the hypersensitive response in tobacco caused by Erwinia amylovora, elicits active oxygen production in suspension-cultured cells. Plant Physiol 102: 1341–1344 (1993).
Bolwell GP: Dynamic aspects of the plant extracellular matrix. Int Rev Cytol 146: 261–324 (1993).
Bolwell GP: A role for phosphorylation in the down-regulation of phenylalanine ammonia-lyase in suspension-cultured cells of French bean. Phytochemistry 31: 4081–4086 (1992).
Bolwell GP, Mavandad M, Millar DJ, Edwards KJ, Schuch W, Dixon RA: Inhibition of mRNA levels and activities by trans-cinnamic acid in elicitor-treated bean cells. Phytochemistry 27: 2109–2117 (1988).
Bolwell GP, Coulson V, Rodgers MW, Murphy DL, Jones D: Modulation of the elicitation response in cultured French bean cells and its implication for the mechanism of signal transduction. Phytochemistry 30: 397–405 (1991).
Bradley DJ, Kjellbom P, Lamb CJ: Elicitor-and wound-induced oxidative cross-linking of a proline-rich plant cell wall protein: A novel rapid defence response. Cell 70: 21–30 (1992).
Carver KA, Walker DA: The effect of adenylate kinase activity in a reconstituted chloroplast system; determination of ATP, ADP and AMP levels with luciferin-luciferase. Sci Tools 30: 1–5 (1983).
Cleveland DW, Fischer SG, Kirschner MW, Laemmli UK: Peptide mapping by limited proteolysis in sodium dodecyl sulphate and analysis by gel electrophoresis. J Biol Chem 252: 1102–1106 (1977).
Constabel CP, Matton DP, Brisson N: Concurrent synthesis and degradation of alcohol dehydrogenase in elicitor-treated and wounded potato tubers. Plant Physiol 94: 887–891 (1990).
Davis D, Merida J, Legendre L, Low PS, Heinstein P: Independant elicitation of the oxidative burst and phytoalexin formation in cultured plant cells. Phytochemistry 32: 607–611 (1993).
Dennis ES, Olive M, Dolferus R, Millar A, Peacock WJ, Setter TL: Biochemistry and molecular biology of the anaerobic response. In: Wray JL (ed), Inducible Plant Proteins, pp. 231–245. Cambridge University Press, Cambridge, UK (1992).
Devlin WS, Gustine DL: Involvement of the oxidative burst in phytoalexin accumulation and the hypersensitive reaction. Plant Physiol 100: 1189–1195 (1992).
Dixon RA, Lamb CJ: Stimulation of de novo synthesis of L-phenylalanine ammonia-lyase in relation to phytoalexin accumulation in Colletotrichum lindemuthianum elicitor-treated cell suspension cultures of French bean (Phaseolus vulgaris). Biochim Biophys Acta 586: 453–463 (1979).
Dixon RA, Lamb CJ: Molecular communication in interactions between plants and microbial pathogens. Annu Rev Plant Physiol Plant Mol Biol 41: 339–367 (1990).
Doke N: Involvement of superoxide anion generation in the hypersensitive response of potato tuber tissues to infection with an incompatable race of Phytophthora infestans and to hyphal wall components. Physiol Plant Path 23: 345–357 (1983).
Farmer EE, Ryan CA: Octadecanoid precursors of jasmonic acid activate the synthesis of wound-inducible protease inhibitors. Plant Cell 4: 129–134 (1992).
Felder MR, Scandalios JG, Liu EH: Purification and partial characterisation of two genetically defined alcohol dehydrogenase isoenzymes in maize. Biochim Biophys Acta 318: 149–159 (1973).
Felix G, Grosskopf DG, Regenass M, Basse CW, Boller T: Elicitor induced ethylene biosynthesis in tomato cells. Plant Physiol 97: 19–25 (1991).
Golden S, Katz J: The determination of reduced nicotinamide adenine dinucleotide and metabolic intermediates in pmol amounts with bacterial luciferase. Biochem J 188: 799–805 (1980).
Kurosaki F, Nishi A: Stimulation of calcium influx and calcium cascade by cyclic AMP in cultured carrot cells. Arch Biochem Biophys 302: 144–151 (1993).
Kurosaki F, Kaburaki H, Nishi A: Synthesis and degradation of cyclic AMP in cultured carrot cells treated with forskolin. Arch Biochem Biophys 303: 177–179 (1993).
Legendre L, Rueter S, Heinstein PF, Low PS: Characterisation of the oligogalacturonide-induced oxidative burst in cultured soybean (Glycine max) cells. Plant Physiol 102: 233–240 (1993).
Lindner WA, Hoffmann C, Grisebach H: Rapid elicitorinduced chemiluminescence in soybean cell suspension cultures. Phytochemistry 27: 2501–2503 (1988).
Llewellyn DJ, Finnegan EJ, Ellis JG, Dennis ES, Peacock WJ: Structure and expression of an alcohol dehydrogenase 1 gene from Pisum sativum (cv. Greenfeast). J Mol Biol 195: 115–123 (1987).
Matton DP, Constabel CP, Brisson N: Alcohol dehydrogenase gene expression following eliclitor and stress treatment. Plant Mol Biol 14: 775–783 (1990).
O'Farrell PH: High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250: 4007–4021 (1975).
Okimoto R, Sachs MM, Porter EK, Freeling M: Patterns of polypeptide synthesis in various maize organs under anaerobiosis. Planta 150: 89–94 (1980).
Raskin I: Role of salicylic acid in plants. Annu Rev Plant Physiol Plant Mol Biol 43: 439–463 (1992).
Ricard B, Mocquot B, Fournier A, Delseny M, Pradet A: Expression of alcohol dehydrogenase in rice embryos under anoxia. Plant Mol Biol 7: 321–329 (1986).
Robinson PS, Cooke CJ, Newton RP, Walton TJ, Smith CJ: Cyclic nucleotide phosphodiesterase activity of Medicago sativa L. Biochem Soc Trans 20: 355 (1992).
Ryan CA, Farmer FE: Oligosaccharide signals in plants: A current assessment. Annu Rev Plant Physiol Plant Mol Biol 42: 651–674 (1991).
Sachs MM, Freeling M, Okimoto R: The anaerobic proteins of maize. Cell 20: 761–767 (1980).
Sanchez LM, Doke N, Kawakita K: Elicitor-induced chemiluminescence in cell suspension cultures of tomato, sweet pepper and tobacco plants and its inhibition by suppressors from Phytophthora spp. Plant Sci 88: 141–148 (1993).
Schwacke R, Hager A: Fungal elicitors induce a transient release of active oxygen species from cultured spruce cells that is dependent upon Ca2+ and protein kinase activity. Planta 187: 136–141 (1992).
Segal AW, Abo A: The biochemical basis of the NADPH oxidase of phagocytes. Trends Biochem Sci 43–47 (1993).
Showalter AM, Bell JN, Cramer CL, Bailey JA, Varner JE, Lamb CJ: Accumulation of hydroxylation-rich glycoprotein mRNAs in response to fungal elicitor and infection. Proc Natl Acad Sci USA 82: 6551–6553 (1985).
Stab MR, Ebel J: The effects of Ca2+ on phytoalexin induction by fungal elicitor in soybean cells. Arch Biochem Biophys 257: 416–423 (1987).
Walton TJ, Cooke C, Newton RP, Smith CJ: Evidence that generation of inositol 1,4,5-triphosphate and hydrolysis of phosphatidylinositol 4,5-bisphosphate are rapid responses following addition of fungal elicitor which induces phytoalexin synthesis in lucerne (Medicago sativa) suspension culture cells. Cell Signalling 5: 345–356 (1993).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Robertson, D., Davies, D.R., Gerrish, C. et al. Rapid changes in oxidative metabolism as a consequence of elicitor treatment of suspension-cultured cells of French bean (Phaseolus vulgaris L.). Plant Mol Biol 27, 59–67 (1995). https://doi.org/10.1007/BF00019178
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00019178