Production of secondary metabolites in plant tissue cultures The goal of this study is to determine the influence of sodium molybdate and sodium tungstate as elicitors on production of scopoletin in cell suspension culture of Angelica archangelica L. The culture was grown in a liquid culture medium Murashige and Skoog on a roller apparatus in the dark and light. The content of scopoletin was in cells and in the culture medium determined by high performance liquid chromatography with fluorometric detection. The results show that sodium molybdate served as an elicitor to the production of scopoletin positively, application of sodium tungstate did not increase scopoletin production in any case. The highest production of scopoletin after application of sodium molybdate versus the control cells was reached in the suspension culture of Angelica archangelica L. cultured in the dark at a concentration of 25,50 mg/l. Scopoletin content increased by 166,7 %. After application of sodium tungstate the production always decreased, in the medium of a suspension culture of Angelica archangelica L. cultivated in the light at a concentration of 66.00 mg/l, the production decrease was by up 62,5 %.

An in vitro system of generating protoplasts from their callus cultures was established. The friable callus was more productive in terms of producing protoplasts than the green compact callus. The concentration of the various cell wall degrading enzymes had an effect on the viability of the protoplasts in the medium. The protoplast system developed from the experiments was stable and could be used for the transformation experiments of Albizia lebek and for other plant improvement practices.

Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmacognosy Candidate: Adriana Blahušová Supervisor: PharmDr. Marie Kašparová, Ph.D. Title of diploma thesis: The study of secondary metabolites in plant tissue cultures I A principal precondition for successful elicitation used to increase the production of secondary metabolites is, among other, finding a suitable elicitor, its concentration and the optimal period of time of the action of the elicitor on the plant culture in vitro, which was the aim of the present diploma thesis. The effect was examined of a 6, 24, 48 and 168 hour action of the solution of chitosan (in four concentrations) on the production of flavonoids in the suspension culture Trifolium pratense L. (variety Sprint, Tempus and DO-8). The culture was cultivated in Gamborg medium to which 2 mg/l of 2,4- dichlorophenoxyacetic acid and 2 mg/l of 6-benzylaminopurine were added, at the temperature of 25řC and 16 hours light/8 hours dark period. The best elicitation effect of chitosan on the production of flavonoids was the lowest concentration of 1 mg/100ml in all three studied varieties after 48 hours of application.