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    Analysis of metabolic variations throughout growth and development of adventitious roots in Silybum marianum L. (Milk thistle), a medicinal plant
    (Springer, 2015) Khan, Mubarak Ali; Abbasi, Bilal Haider; Shah, Naseer Ali; Yücesan, Buhara; Ali, Huma
    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 alpha-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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    Evaluation of biochemical markers during somatic embryogenesis in Silybum marianum L.
    (Springer Heidelberg, 2016) Abbasi, Bilal Haider; Ali, Huma; Yücesan, Buhara; Saeed, Sabahat; Rehman, Khalid
    In present report effects of explants type, basal media and plant growth regulators (PGRs) were tested for induction of indirect somatic embryogenesis in medicinally important plant Silybum marianum L. Leaf, petiole and root explants were exploited in vitro on B5 (Gamborg), SH (Schenk and Hildebrandt) and MS (Murashige and Skoog) media for induction of embryogenic callus followed by somatic embryogenesis. Highest callus induction frequency (76 +/- 4.8 %) was recorded when petiole explants of in vitro derived plantlets were cultured on B5 medium supplemented with 1.5 mg l(-1) 2,4-dichlorophenoxyacetic acid (2,4_D) in combination with 1.5 mg l(-1) Thidiazuron (TDZ). Induction and multiplication of somatic embryos were observed, when the embryogenic calluses were sub-cultured on to B5 medium containing 0.5 mg l(-1) 2,4-D plus 1.5 mg l(-1) TDZ. At this PGRs treatment, 77 % of the cultures responded with 39.1 somatic embryos per callus. Furthermore, MS0 medium was indicated more reponsive for growth and maturation of somatic embryos. Analysis of biochemical markers during various growth phases in somatic embryogenesis revealed that somatic embryos exhibited highest level of total carbohydrate, starch, ascorbic acid and total free amino acids. However, higher protein levels were detected in non-embryogenic callus. Nevertheless, considerable amount of silymarin (4.1 mg g(-1) DW) was detected in somatic embryos than other growth phases. Thus, the present study concluded that biochemical and physiological changes during embryogenesis are influenced by interplay of explants type, basal media and PGRs.