| Summary: | 碩士 === 國立屏東科技大學 === 熱帶農業暨國際合作系所 === 96 === Pharmacologically, Radix Sophorae flavescentis is one of the most broadly used medicinal raw material in Oriental Medicine. Advanced biotechnological methods of culturing plant cells and tissues provides new means of conserving and rapidly propagation of this valuable endangered medicinal plant. In this report, micro-propagating system based on the seeds and young stem node segments of Sophora flavescens Ait (Fabaceae) was established. The effect of in vitro tissue propagated source of the plant on matrine alkaloids were investigated by reversed phase HPLC. Murashige and Skoog (MS) basal medium based on Gelrite at 0.18% has the higher the growth responses than MS basal medium based on Agar 0.7% without plant growth regulators (PGRs) and then MS basal mediums supplemented with PGRs. Results shown MS basal medium based on Gelrite at 0.18% 1ppm IBA induced maximum shoot growth with 84.4± 3.3 rooting from the base of the regenerated shoot, which was higher than 76.9 ± 3.4 rooting of Agar at 0.7% supplemented with 1ppm IBA. Among the different concentrations of Gelrite tested, maximum proliferation rate was recorded at 0.18% of Gelrite. Combination of Agar and Gelrite tested by four variations. The combinations reduced the proliferation rate. The main alkaloid of S. flavescens were analyzed reversed phase C-18 column by gradient elution using 0.01% acetic acid–acetonitrile (60:40 v/v) as mobile phase. Calibration graphs were constructed in the range 0.2–27.7 µg/ml for matrine. The linearity of the plot concentrations (x, µg/ml) for each matrine alkaloid versus peak areas (y) was investigated; the results are expressed as the values of the determination of coefficient (R2). Good linear relationships were showed with correlation coefficients for the analytes exceeded 0.992. Intra-day assay and inter-day assay precision of the analytes were less than 2.97%. HPLC analysis showed that the samples all contained quinolizidine alkaloids and a main alkaloid chromatographic peaks were identified by referring to standard compounds. Among six S. flavescens alkaloids, the content of matrine is the highest in all of the samples. Matrine alkaloids were slightly different between the processed S. flavescens roots in pod and field, and two different MS medium, with a character of that matrine has a higher content than the others. Higher content of matrine is obtained in the wild roots, which is slightly different from commercial one and also higher in asexual propagation with Gelrite solidified media than Agar solidified media.
|
|---|
