论文部分内容阅读
目的以具有产石杉碱甲能力的蛇足石杉离体叶状体为研究对象,探究蛇足石杉离体培养物累积石杉碱甲的影响因素及生产动态变化规律。方法应用正交设计对蛇足石杉离体培养叶状体的植物生长物质浓度进行筛选,采用HPLC法对叶状体提取物中的目标成分进行定性定量检测,用Logistic模型拟合叶状体的生长曲线。结果以白光为主要光源,光照时间为12 h/d,1/4 MS固体培养基,为蛇足石杉叶状体产石杉碱甲的最佳条件,相对增殖率达到3 174.5%,石杉碱甲累积量为72.45μg/L;蛇足石杉离体叶状体增殖的生长曲线呈“S”形,继代周期以50 d左右为最适;60 d后生长进入稳定阶段,累积石杉碱甲量明显加快,直至85 d达到最大值7.453μg/g,即叶状体先快速增殖,后累积石杉碱甲。结论叶状体累积石杉碱甲与叶状体增殖呈非生长偶联型,叶状体增长规律符合Logistic生长模型,最大的比生长速率(μmax)为0.071·d-1。
OBJECTIVE: To study the influencing factors and the dynamic changes of huperzine A accumulation in the isolated culture of Huperzia serrata in vitro, and to study the mechanism of the production of huperzine A in vitro. Methods The orthogonal design was used to screen the concentration of phytohormones in vitro of Phyllostachys heterocycla cv. Phyllostachys pubescens. The target components in the extracts of phyllodes were qualitatively and quantitatively determined by HPLC. Growth curve. Results White light as the main light source, light time of 12 h / d, 1/4 MS solid medium, the best conditions for the Huperzia serrata leaf phytate, the relative proliferation rate reached 3 174.5% The accumulation of alkali and alkaloids was 72.45μg / L. The growth curve of detached fronds from the genus Hippeastrum was “S” shape, and the subculture cycle was about 50 days. The growth was stable after 60 days Huperzine A significantly accelerated until 85 d reached a maximum of 7.453μg / g, that the rapid proliferation of fronds after the first accumulation of huperzine A. Conclusions The accumulation of huperzine A and the proliferation of the fronds in the fronds was non-growth-coupled. The growth pattern of the fronds accorded with the Logistic growth model. The maximum specific growth rate (μmax) was 0.071 · d-1.