Regulacija produkcije labdanskih diterpena u kulturi in vitro Cistus creticus L. i analiza njihove biološke aktivnosti
Regulation of labdane diterpenes production in Cistus creticus L. in vitro culture and analysis of their biological activity.
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Cistus creticus L. is a perennial, bushy plant that inhabits mainly dry and warm areas of the Mediterranean. The resin of this species has been actively collected by the local inhabitants of Crete since ancient times due to its aromatic and medicinal properties. The predominant bioactive component of the resin and the shoots of this plant are labdane diterpenes, while the chemical composition of the roots is still unknown. In order to find the optimal conditions for the labdane diterpenes production, the subject of this thesis was to investigate the regulation of labdane-type diterpenes production in in vitro grown plants of C. creticus, as influenced by various abiotic factors and growth media with varying content of different carbohydrates. In vitro culture of C. creticus was successfully established on hormone-free MS medium, and the parameters of growth and development of explants showed no significant differences among analyzed clones. Optimal concentrations of carbohydrates for in vitro growth and development of C. creticus ranged from 0.03 M to 0.1 M sucrose or glucose. Genetic transformation of C. creticus was performed with Agrobacterium rhizogenes A4M70GUS and the culture of transformed roots was established. Regeneration of transformed plants was unsuccessful. Histochemical analysis of the leaves from in vitro cultured C. creticus shoots showed that labdane diterpenes and phenolic compounds accumulate in the type II capitate glandular trichomes. Phytochemical characterization of in vitro grown C. creticus has enabled the identification and quantification of two major groups of secondary metabolites (terpenes and phenols), which are produced under these conditions. Predominant compounds accumulating in shoots of C. creticus grown in vitro were labdane diterpenes, the most abundant among them being labd-7,13-diene-15-ol and labd-7,13-dien-15-ol acetate. Amongst the volatile secondary metabolites, the most abundant were monoterpenes and sesquiterpenes. Labd-7,13-dien-15-ol acetate was the most abundant compound in dichloromethane extracts of type II capitate trichomes exudates. In the roots of C. creticus, the presence of the phenolic compounds and fatty acids was determined, whereas labdane diterpenes were not detected...
Keywords:Cistus creticus L.; Labdane diterpenes; In vitro culture; Phenolic compounds; MVA and MEP biosynthetic pathway; Antioxidant activity; Antimicrobial activity; Cytotoxic activity
Source:University of Belgrade, Faculty of Biology, 2014, 1-207
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