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Effect of exogenously applied jasmonic acid (JA) in combination with calcium and verapamil (a calcium channels blocker) on the production of flavonoids and isoflavonoids in suspension cultures of Trifolium pratense L. was investigated. The culture was cultivated in Gamborg medium with an addition of 2 mg.l(-1) of 2,4-dichlorophenoxyacetic acid and 2 mg.l(-1) of 6-benzylaminopurine, at the temperature of 25 °C, 16-hr light/8-hr dark period. The best effect of jasmonic acid on the production of flavonoids and isoflavonoids was manifested after a 24-hour application of the 50 μmol.l(-1) concentration. The maximum production of JA-induced suspension culture was observed when cells were treated with a high level of calcium (10 mmol.l(-1)). The addition of all concentrations of verapamil to JA-induced suspension culture decreased production of flavonoids and isoflavonoids.

The plant cell may respond to the excess of heavy metals in its environment by various mechanisms, including enhanced biosynthesis of secondary metabolites. In this study, zinc (0 to 1500 μM) and cadmium ions (0 to 100 μM) were tested as potential elicitors of the production of coumarins in angelica cell suspension cultures. In addition, the toxicity of both metals was assessed by evaluating their effect on cell growth (characterized by fresh and dry biomass at the end of a two-week subculture). It has been found that fresh biomass was not influenced up to zinc concentrations of 150 and 300 μM in the dark-grown and light-grown cultures, resp. Then it declined with an increasing zinc level. Zinc at 1500 μM diminished it by 54% and 24% in the dark-grown and light-grown cultures, resp. Dry biomass was influenced in a similar way. Zinc at 1500 μM reduced dry cell weight by 30% and 20% in cultures in the dark and in the light, resp. Cadmium ions did not affect fresh and dry weights of cells up to concentrations of 10 μM and 50 μM in cultures in the dark and in the light, resp. Toxic concentrations of cadmium are by an order of magnitude lower than those of zinc. Cadmium at 50 μM reduced fresh and dry cell weights by 66% and 59%, resp., in the dark-grown cultures. Cadmium at 100 μM caused a decrease in fresh and dry biomass by 40% and 44%, resp., in the light-grown cultures. Neither zinc nor cadmium improved production of coumarins.