雷公藤不定根两相培养初步研究

Preliminary Study on Two-phase Culture of Tripterygium wilfordii Adventitious Root

以雷公藤(Tripterygium wilfordii)不定根为材料,通过两相培养技术,研究有机溶剂邻苯二甲酸二丁酯(DBP)不同浓度及培养时间对雷公藤不定根生长及次生代谢产物含量的影响。结果显示,DBP浓度为6%、培养至第6 d时,不定根增长量达1.14 g/瓶,为对照(1.08 g/瓶)的1.06倍;DBP浓度为2%、培养至第8 d时,内酯醇含量达74.96μg/g,为对照(53.67μg/g)的1.40倍;DBP浓度为2%、培养至第2 d时,所收获的内酯醇总产量最高(0.40 mg/瓶),且为对照(0.27 mg/瓶)的1.48倍。本研究结果表明,在培养基中添加一定浓度DBP,虽然适合雷公藤内酯醇的形成,但不适合雷公藤生物碱的形成;不论添加DBP浓度大小、培养时间长短,不定根中吉碱和次碱含量及每瓶总产量均低于对照。

Through two-phase culture,we studied the effects of culture time extension and volume percentages of dibutylphthalate（ DBP） on growth and secondary metabolite content in the adventitious roots of Tripterygium wilfordii Hook. f.. Results showed that adventitious root growth increased 1. 06-fold when the volume percentage of DBP was 6% and addition of organic solvent was on day 6. Triptolide content in the root was 1. 40-fold higher when the volume percentage of DBP was 2% and addition of organic solvent was on day 8. Triptolide yield in the two-phase culture was 1. 48-fold higher than that of the control when the volume percentage of DBP was 2% and addition of organic solvent was on day 2. Thus,DBP in the medium was suitable for triptolide accumulation, but alkaloid accumulation was inhibited significantly. The contents of wilforgine and wilforine in the adventitious roots and yields of wilforgine and wilforine per flask were all lower than that of the control,regardless of the concentration of DBP added to the medium or the extension of culture time.