Regeneration of Cichorium intybus L. in tissue culture and analysis of its secondary metabolites

Cichorium intybus L. was collected from field and regenerated in vivo in soil and in vitro in three different media; Murashige and Skoog (MS), Gamborg B5 and White medium supplemented with different concentration and combinations of auxins and cytokinins. Callus production was also achieved in the basic media supplemented with plant growth regulators (PGRs). Plant was regenerated by direct organogenesis from leaf explant in MS and B5 medium supplemented with Indole-3-butyric acid (IBA), and Naphthalene acetic acid (NAA). Root and shoot regeneration occurred. Plantlets were maturated in growth chamber in sterile condition and acclimatized in laboratory in ambient conditions and transferred to soil. Callus was produced from leaf explant in MS, B5 and White medium supplemented with auxin, cytokinin and gibberellic acid. The best result for callus production was provided from MS medium supplemented with Indole-3-acetic acid IAA and Benzyl amino purine (BAP). Secondary metabolites provided from in vivo and in vitro samples were analysed by High Performance Liquid Chromatography (HPLC) and Spectrophotometry. Phenolic compounds (esculine, caftaric acide, chlorogenic acid and cichoric acid) were extracted in ethanol/water (80:20 v/v) and quantified by HPLC. Inulin was quantified by spectrophotometry. The highest caftaric acid concentration was found in flowers from in vivo samples. However, the highest concentration of esculine was found in leaf extract of in vitro regenerated plants. Chlorogenic acid concentration was higher in vivo samples then that of in vitro whereas chichoric acid concentration was found to be higher in in vitro samples than that of in vivo. Inulin was found to be higher in in vivo samples.