A very efficient and rapid regeneration system via multiple shoot formation was developed for Cichorium intybus L. when leaf explants excised from sterile seedlings were cultured on medium supplemented with different concentrations and combinations of various plant growth regulators. In a comparison of leaf lamina and petiole explants, lamina explants produced over three times more shoots than petiole explants, with a mean of 7.5 shoots compared to 2.4. Of the combinations of KIN/ IAA, KIN/NAA, BAP/IAA, or BAP/NAA, 0.5 mg l -1 KIN combined with 0.3 mg l -1 IAA was the most effective, producing a mean of 19.7 shoots per lamina explant while the control treatment involving no plant growth regulators produced no shoots at all. When either cytokinin was used alone, BAP was found nearly twice more successful than KIN. However, the most effective treatment of all was the combination of 0.01 mg l -1 TDZ and 1.0 mg l -1 IAA, producing as many as 35.8 shoots per lamina explant. This rate of shoot regeneration is remarkably higher than those previously reported for C. intybus, most likely due to the highly inductive effect of TDZ, which was tested for the first time in this species. Rooting of the shoots was readily achieved on medium containing different concentrations of IAA or IBA. IAA was more effective than IBA and resulted in the highest frequency of shoots that rooted (100%) and mean number of roots per shoot (4.2) when used at 0.5 mg l -1 . Hardening off process resulted in a production of more than 80% healthy plantlets.
Silybum marianum L. is a medicinal plant used in the treatment for jaundice and liver diseases. In this study, an adventitious root culture was developed for the production of health promoting phytochemicals. Adventitious roots were induced from nodal explants on solid Murashige and Skoog medium supplemented with 1.0 mg l -1 of a-Naphthalene acetic acid. Growth kinetics of the roots was investigated every week, for 8 weeks of culture period. Highest fresh biomass formation (153 mg l -1 ) was observed in 6-week old cultures. Adventitious roots were harvested from different growth stages as control (CTR), lag phase (LAG), logarithmic phase (LOG) or stationary phase (STN). Metabolite profiling of the samples was investigated using electro spray ionization time of flight mass spectrometry. Significant phenylpropanoids such as cinnamic acid and di-hydro kaempferol were predominantly found in LOG phase, whereas the highest amount of malonic acid was detected in STN as compared to other growth phases. More sucrose content was detected in CTR, while the tryptophan content was higher in LOG phase. Among the vital fatty acids, prostaglandin A1 and phenyl acetic acid were at highest levels in STN phase. However, more brassicasterols were observed in LAG phase than other growth phases. Punicic acid and lignan pinoresinol were detected abundantly in the LOG phase. Biochemical characterization revealed significant correlations between silymarin content and DPPH as well as TPC and TFC in the growth curve. Interestingly, among all growth stages there was no correlation of PAL activity with TFC and silymarin content.
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