Production of secondary metabolites in the explant cultures of Trichocereus pachanoi (Cactaceae) Lucie Urbanová Diploma thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Pharmacy Key words: Trichocereus pachanoi, the explant cultures, mescaline, secondary metabolites, prodrugs, elicitation, cinnamic acid, sodium cinnamate, tryptophan, ozone, methylene blue The aim of this thesis was the research of effect of selected precursors and elicitors for secondary metabolite production of mescaline in explant cultures Trichocereus pachanoi. Suspension cultures were cultivated on the Murashige and Skoog medium with addition of the growth regulators 2,4-dichlorophenoxyacetic acid and 6-benzylaminopurine. To observe changes in production these precursors: cinnamic acid, sodium cinnamate and tryptophan, and these elicitors: ozone and methylene blue, were selected. Precursors were prepared and used in concentrations of 50, 100 and 500 mg·l-1. Ozone was added in two time slots, 5 and 10 seconds. Methylene blue has been used in concentrations of 1, 10 and 100 mg·l-1. Cultivation with added compounds lasted for 24 and 168 hours. The content of mescaline was determined by HPLC analysis. A statistically significant effect on the production of mescaline in vitro culture was proved only with...

Affecting tissue cultures with precursors is one of methods that increases the production of secondary metabolites in vitro. This thesis deals with precursors affecting production of mescaline in suspension tissue cultures of Trichocereus pachanoi. As precursors dopamine, D, L-tyrosine, casein hydrolysate and shikimic acid were used. Three concentrations of these precursors were prepared. Suspension cultures with different concentrations of precursors were analysed by HPLC method after 48 and 168 hours. Murashige and Skoog medium was used for cultivation. Mescaline biosynthesis was most affected by dopamine. The largest amount of mescaline was produced in suspension culture with the highest concentration of dopamine 50 mg/100 ml.The culture was cultivated for 168 hours. Suspension cultures that were cultivated for 48 hours produced the highest amount of mescaline with the highest concentration of dopamin 50 mg/100 ml. The biosynthesis of mescaline was inhibited by other precursores (D, L-tyrosine, casein hydrolysate, shikimic acid).