不同发育期菘蓝和毛状根中直铁线莲宁B的含量变化规律研究

Dynamic Analysis of Clemastanin B of the Plant at Different Growth Stages and Hairy Roots of Isatis indigotica

目的:测定不同生长时期菘蓝及不同悬浮培养时间毛状根中抗流感病毒活性成分直铁线莲宁B的含量,研究其累积变化规律,为菘蓝的采收和菘蓝毛状根体系评价提供参考。方法:应用超高效液相色谱测定菘蓝叶片、根及毛状根中直铁线莲宁B的含量。结果:菘蓝叶片中未检测到直铁线莲宁B;在整个生育期内,菘蓝根中直铁线莲宁B动态累积变化曲线为"单峰型",含量在生长100 d左右达到峰值后逐渐降低;毛状根中直铁线莲宁B含量随悬浮培养时间持续增加,在20 d达到5.40 mg·g^(-1),超过根中最大含量。结论:整个生长周期内,菘蓝和毛状根中直铁线莲宁B累积变化呈一定规律性,原植物中直铁线莲宁B在九月下旬产量达到最大;毛状根在10~20 d期间直铁线莲宁B含量增长处于生长对数期,20 d后即超过原植物的最大含量,说明菘蓝毛状根体系是研究直铁线莲宁B生物合成的理想体系,也可为后续利用菘蓝毛状根体系大量制备直铁线莲宁B提供参考。

Objective： To investigate the changes of clemastanin B of the plant at different growth stages and hairy root of lsatis indigotica. Methods： Ultra Performance Liquid Chromatography was applied to determine the content of clemas- tanin B from the plant and hairy roots of I. indigotica. Results ： Clemastanin B was undetectable in the leaves of I. indigotica, and the content of clemastanin B exhibited obvious change across different growing stages and increased with culture time in hairiry roots. The content of clesmastanin B showed a continuous increasing trend in hairy roots, reaching 5.40 mg · g^ -1 in 20 days, which exceeding the maximum content of clemastanin B in roots, while its content was gradually decreased after reaching the peak at 100 days. Conclusion： Investigation of clemastanin B content at t different growth stages and hairy roots in I. indigotica present a certain law. The content of clemastanin B in the roots reached the maximum in 100 days after seeding. However, the content of clemastanin B in hairy roots were at the logarithmic phase of growth from 10 days to 20 days, even exceeding the maximum content in the roots at 20 days after seeding. From this angle, the hairy roots may provide us an i- deal system to dissect the mechanism of the biosynthesis of clemastanin B and produce clemastanin B.