Abstract
Stilbene synthase is an inducible enzyme occurring in a small number of plants. The enzyme is amenable to analysis and biochemical studies only after the cells are subjected to induction. Cell suspension cultures of peanut react very selectively if elicited with biotic inducers. Just as intact peanut plants produce stilbene phytoalexins when attacked by fungi so also do sterile cultured cells when treated with sterilized insoluble fungal cell walls. Both systems react by synthesizing stilbene synthase. The time courses of increase in enzyme activity, protein synthesis and mRNA activity were studied, and their relation to other activities of the cells was elaborated. The results show that, after applying the fungal elicitor, the system responds very quickly and selectively: within 2 hours the synthesis rate of stilbene synthase protein is increased more than 30-fold, the increase being detectable 40 min after induction. The first increase in translatable mRNA for stilbene synthase can be seen 20 min after application of the stimulus. Stilbene synthase synthesized in vivo was compared to stilbene synthase prepared by translation in vitro. There was no difference in size, and limited proteolysis did not indicate significant differences in the peptide structure of the primary translation product and the active enzyme.
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References
Booth C: Fungal culture media. In: Booth C (ed.) Methods in Microbiology Vol. 4. Academic Press, London, New York (1971) pp 49–94.
Cleveland DW, Fischer ST, Kirschner MW, Laemmli UK: Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem 252: 1102–1106 (1977).
Ebel J: Phytoalexin synthesis: The biochemical analysis of the induction process. Annu Rev Phytopathol 24: 235–264 (1986).
Fritzemeier K-H, Kindl H: Coordinate induction by UV-light of stilbene synthase, phenylalanine ammonia-lyase and cinnamate 4-hydroxylase in leaves of Vitaceae. Planta 151: 48–52 (1981).
Fritzemeier K-H, Kindl H: 9,10-Dihydrophenanthrenes as phytoalexins of Orchidaceae. Eur J Biochem 133: 545–550 (1983).
Fritzemeier K-H, Rolfs C-H, Pfau J, Kindl H: Action of UV-C on stilbene formation in callus of Arachis hypogaea. Planta 159: 25–29 (1983).
Gamborg OL, Miller RA, Ojima K: Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50: 151–158 (1968).
Gerdes H-H, Kindl H: Partial purification of mRNAs encoding glycollate oxidase and catalase. Planta 167: 166–174 (1986).
Kindl H: Biosynthesis in stilbenes. In: Higuchi T (ed.) Biosynthesis and Biodegradation of Wood Compounds. Academic Press, New York (1985) pp. 349–377.
Kreuzaler F, Ragg H, Heller W, Tesch R, Witt I, Hammer D, Hahlbrock K: Flavanone synthase from Petroselinum hortense. Eur J Biochem 99: 89–96 (1979).
Kuhn DN, Chappell J, Boudet A, Hahlbrock K: Induction of phenylalanine ammonia-lyase and 4-coumarate: CoA ligase mRNA in cultured plant cells by UV light and fungal elicitor. Proc Natl Acad Sci USA 81: 1102–1106 (1984).
Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685 (1970).
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275 (1951).
O'Farrell PH: High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250: 4007–4021 (1975).
Rolfs CH, Fritzemeier K-H, Kindl H: Cultured cells of Arachis hypogaea susceptible to induction of stilbene synthase (Resveratrol forming). Plant Cell Reports 1: 83–85 (1981).
Rolfs C-H, Kindl H: Stilbene synthase and chalcone synthase. Two different constitutive enzymes in cultured cells of Picea excelsa. Plant Physiol 75: 489–492 (1984).
Rolfs C-H, Schön H, Steffens M, Kindl H: Cell suspension culture of Arachis hypogaea: Model system of specific enzyme induction in secondary metabolism. Planta 172: 238–244 (1987).
Ryder TB, Cramer CL, Bell JN, Robbins MP, Dixon RA, Lamb CJ: Elicitor rapidly induces chalcone synthase mRNA in Phaseolus vulgaris cells at the onset of the phytoalexin defense response. Proc Natl Acad Sci 81: 5724–5728 (1984).
Schenk RU, Hildebrandt AC: Medium and techniques for induction and growth of monocotyledonous and dicotyledonous plant cell cultures. Can J Bot 50: 199–204 (1972).
Schöppner A, Kindl H: Stilbene synthase (Pinosylvin synthase) and its induction by UV-light. FEBS Lett 108: 349–352 (1979).
Schöppner A, Kindl H: Purification and properties of a stilbene synthase from induced cell suspension cultures of peanut. J Biol Chem 259: 6806–6811 (1984).
Schröder J, Kreuzaler F, Schäfer E, Hahlbrock K: Concomitant induction of phenylalanine ammonia-lyase and flavanone synthase mRNAs in irradiated plant cells. J Biol Chem 254: 57–65 (1979).
Stephan D, vanHuystee RB: Some aspects of peroxidase synthesis by cultured peanut cells. Z Pflanzenphysiol 101: 313–322 (1981).
Stoessl A: Biosynthesis of phytoalexins. In: Bailey JA, Mansfield JW (eds) Phytoalexins. Blackie, Glasgow (1982) pp 133–180.
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Vornam, B., Schön, H. & Kindl, H. Control of gene expression during induction of cultured peanut cells: mRNA levels, protein synthesis and enzyme activity of stilbene synthase. Plant Mol Biol 10, 235–243 (1988). https://doi.org/10.1007/BF00027400
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DOI: https://doi.org/10.1007/BF00027400