Abstract
Approximately fourfold increases in the extractable activity of the enzyme chalcone isomerase (CHI, EC 5.5.1.6) were observed within 24 h of treatment of cell suspension cultures of Phaseolus vulgaris with a crude elicitor preparation heatreleased from the cell walls of the bean pathogen Colletotrichum lindemuthianum. The induction of CHI activity was highly dependent upon elicitor concentration, with maximum induction occurring in two discrete concentration ranges. A basal half-life for CHI>32 h in control cultures was determined by labelling with 2H from 2H2O followed by analysis of the equilibrium distribution of enzyme activity in CsCl density gradients. Comparative density labelling indicated that at both the lower and higher effective elicitor concentrations, the induced appearance of CHI activity was the result of an apparent initial activation of pre-existing enzyme followed by an increase in the rate of de-novo synthesis of the enzyme as compared with non-elicited controls. The increased appearance of the enzyme over the first 8 h in elicitor-treated cultures was inhibited by cycloheximide, cordycepin and actinomycin D. The results are discussed in relation to the mechanisms of co-ordinate enzyme induction operating in French-bean cell cultures exposed to fungal elicitors.
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Abbreviations
- CHI:
-
chalcone isomerase (EC 5.5.1.6)
- CHS:
-
chalcone synthase
- 4CL:
-
4-hydroxycinnamic acid:CoA ligase (FC 6.2.1.12)
- PAL:
-
phenylalanine ammonia-lyase (EC 4.3.1.5)
References
Acton, G.J., Schopfer, P. (1975) Control over activation or synthesis of phenylalanine ammonia-lyase by phytochrome in mustard (Sinapis alba). A contribution to eliminate some misconceptions. Biochim. Biophys. Acta 404, 231–242
Acton, G.J., Drumm, H., Mohr, H. (1974) Control of synthesis de novo of ascorbate oxidase in the mustard seedling (Sinapis alba L.) by phytochrome. Planta 121, 39–50
Boudet, A., Humphrey, T.J., Davies, D.D. (1975) The measurement of protein turnover by density labelling. Biochem. J. 152, 409–416
Dixon, R.A., Bendall, D.S. (1978a) Changes in phenolic compounds associated with phaseollin production in cell suspension cultures of Phaseolus vulgaris. Physiol. Plant Pathol. 13, 283–294
Dixon, R.A., Bendall, D.S. (1978b) Changes in the levels of enzymes of phenylpropanoid and flavonoid synthesis during phaseollin production in cell suspension cultures of P. vulgaris. Physiol. Plant Pathol. 13, 295–306
Dixon, R.A., Lamb, C.J. (1979) Stimulation of de novo synthesis of L-phenylalanine ammonia-lyase in relation to phytoalexin accumulation in Colletotrichum lindemuthianum clicitor-treated cell suspension cultures of French bean (Phaseolus vulgaris). Biochim. Biophys. Acta 586, 453–463
Dixon, R.A., Dey, P.M., Murphy, D.L., Whitehead, I.M. (1981) Dose responses for Colletotrichum lindemuthianum elicitor-mediated enzyme induction in French bean cell suspension cultures. Planta 151, 272–280
Dixon, R.A., Dey, P.M., Whitehead, I.M. (1982) Purification and properties of chalcone isomerase from cell suspension cultures of Phaseolus vulgaris. Biochim. Biophys. Acta 715, 25–33
Fourcroy, P. (1978) Advantages of potassium bromide gradients for isopycnic centrifugation of proteins. Biochem. Biophys. Res. Commun. 84, 713–720
Geissmann, T.A., Clinton, R.O. (1946) Flavanones and related compounds. I. The preparation of polyhydroxy-chalcones and-flavanones. J. Am. Chem. Soc. 68, 697–700
Hahlbrock, K., Grisebach, H. (1979) Enzymic controls in lignin and flavonoid biosynthesis. Annu. Rev. Plant Physiol. 30, 105–130
Hahlbrock, K., Zilg, H., Grisebach, H. (1970) Stereochemistry of the enzymatic cyclisation of 4,2′,4′-trihydroxychalcone to 7,4′-dihydroxyflavanone by isomerases from mung bean seedlings. Eur. J. Biochem. 15, 13–18
Hu, A.S.L., Bock, R.M., Halvorson, H.O. (1962) Separation of labeled from unlabeled proteins by equilibrium density gradient sedimentation. Anal. Biochem. 4, 489–504
Johnson, C.B. (1977) The use of density labeling techniques in investigations into the control of enzyme levels. In: The regulation of enzyme synthesis and activity in higher plants, pp. 225–244, Smith, H., ed. Academic Press, London New York
Johnson, C.B. (1979) Activation, synthesis and turnover of nitrate reductase controlled by nitrate and ammonium in Chlorella vulgaris. Planta 147, 63–68
Kuhn, B., Forkmann, G., Seyffert, W. (1978) Genetic control of chalcone-flavanone isomerase activity in Callistephus chinensis. Planta 138, 199–203
Lamb, C.J., Dixon, R.A. (1978) Stimulation of de novo synthesis of L-phenylalanine ammonia-lyase during induction of phytoalexin biosynthesis in cell suspension cultures of Phaseolus vulgaris. FEBS Lett. 94, 277–280
Lamb, C.J., Rubery, P.H. (1976) Interpretation of the rate of density labelling of enzymes with 2H2O. Possible implications for the mode of action of phytochrome. Biochim. Biophys. Acta 421, 308–318
Lamb, C.J., Bell, J.N., Norman, P.M., Dixon, R.A., Bailey, J.A., Rowell, P.M. (1983) Early molecular events in the phytoalexin defense response. In: NATO-ASI Structure and function of the plant genome, pp. 313–328, Dure, L., Cifferri, O., eds. Plenum Press, New York
Lawton, M.A., Dixon, R.A., Lamb, C.J. (1980) Elicitor modulation of the turnover of L-phenylalanine ammonia-lyase in French bean cell suspension cultures. Biochim. Biophys. Acta 633, 162–175
Lawton, M.A., Dixon, R.A., Hahlbrock, K., Lamb, C.J. (1983a) Rapid induction of the synthesis of phenylalanine ammonia-lyase and of chalcone synthase in elicitor-treated plant cells. Eur. J. Biochem. 129, 593–601
Lawton, M.A., Dixon, R.A., Hahlbrock, K., Lamb, C.J. (1983b) Elicitor induction of mRNA activity. Rapid effects of elicitor on phenylalanine ammonia-lyase and chalcone synthase mRNA activities in bean cells. Eur. J. Biochem. 130, 131–139
Leggett-Bailey, J (1962) In: Techniques in protein chemistry, pp. 293–295. Elsevier, Amsterdam
Sharma, R., Schopfer, P. (1982) Sequential control of phytochrome-mediated synthesis de novo of β-amylase in the cotyledons of mustard (Sinapis alba L.) seedlings. Planta 155, 183–189
Smith, H., Billett, E.E., Giles, A.B. (1977) The photocontrol of gene expression in higher plants. In: Regulation of enzyme synthesis and activity in higher plants: problems and techniques, pp. 93–126, Smith, H., ed. Academic Press, London New York
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Dixon, R.A., Gerrish, C., Lamb, C.J. et al. Elicitor-mediated induction of chalcone isomerase in Phaseolus vulgaris cell suspension cultures. Planta 159, 561–569 (1983). https://doi.org/10.1007/BF00409146
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DOI: https://doi.org/10.1007/BF00409146