代谢中间产物和诱导子对南方红豆杉培养细胞生长和紫杉醇含量的影响

Influence of Metabolic Intermediate Products on Culture Cell Growth and Taxol Content of Taxus chinensis var. mairei

以南方红豆杉 (Taxuschinensisvar.meirei)悬浮细胞为材料 ,利用紫杉醇合成途径中 3 -羟基 -3 -甲基戊二酸还原酶 (HMGR)抑制剂研究其合成途径的多样性 ,以及代谢中间产物 (牛儿醇 ,牛儿牛儿醇 ,蒎烯 ,甲瓦龙酸 )和诱导子 (花生四烯酸 ,茉莉酸甲酯 )对培养细胞生长和紫杉醇含量的影响 ,以探讨紫杉醇生物合成途径人工调控的可能性。结果 :从培养细胞生长曲线看 ,细胞干重和紫杉醇含量均在培养第 30天达到最高 ,HMGR抑制剂compactin对紫杉醇生物合成的抑制作用不明显 ;在培养开始时 ,加入牛儿牛儿醇 (5mg/L)能显著提高紫杉醇含量 ,牛儿醇 (10mg/L)、甲瓦龙酸 (10 0mg/L)对紫杉醇的合成也有一定的促进作用 ,而蒎烯(10mg/L)没有作用 ;加入花生四烯酸 (3 .2 μmol/L)和茉莉酸甲酯 (10 0 μmol/L)分别提高紫杉醇含量 1倍和 10倍。结论 :在南方红豆杉培养细胞中 ,紫杉醇生物合成除了经典的乙酸 /甲羟戊酸途径外 ,还可能存在其他途径。加入合适的代谢中间产物的诱导子可以提高植物培养细胞中的紫杉醇含量 ,但由于其生物合成步骤复杂 。

In order to understand influence of metabolic intermediate products of geraniol, geranyl geraniol, pinene, and hiochic acid on culture cell growth and taxol content of Taxus chinensis var, mairei and to explore the biological synthetic channel and artificial regulation of taxol, suspension cells to Taxus chinensis var. mairei cultured continuously for over six generations were taken as experimental materials for observation. The results showed that the inhibition function of HMGR inhibitor, compactin, for biological synthesis of taxol was not obvious, indicating that beside the channel of typical acetic acid/methylhydroxyl valeric acid synthesis, there might be other channels. Judging from the curve line of culture cell growth, the dry weight of cells and taxol content reached their peak in the thirtieth day. After geranyl geraniol (5 mg/L) was added, taxol content was remarkably increased. There was also certain promoting function in geraniol (10mg/L) and hiochic acid (100mg/L) for taxol synthesis. But there was no any function in pinene. The findings indicate that if the proper metabolic intermediate products were added, it is possible to enhance taxol content in culture cells of the plants. But due to complicated steps in its biological synthesis, the influence of the early metabolic intermediate products is comparatively slight for taxol synthesis.