基于定量PCR技术探讨紫杉醇生物合成的限速步骤

Exploration of the Speed-limiting Process for Taxol Biosynthesis Resorting to Quantitative PCR

次生代谢产物生物合成受到发育和诱导的调控,本实验研究了组织分化和诱导处理对紫杉醇生物合成的影响,并采用定量PCR技术分析了紫杉醇生物合成不同阶段关键酶基因的动态表达特征.结果表明,紫杉醇主要分布在中国红豆杉(T axus ch inensis)树皮和根皮组织中,针叶内含量很少,催化紫杉醇功能官能团连接的关键酶基因也主要定位在树皮和根皮组织中;茉莉酸甲酯(M J)和真菌诱导子F 5分别提高了中国红豆杉悬浮培养细胞HG-1紫杉醇得率8倍和10倍,同时有效诱导紫杉醇生物合成基因的表达,发现催化紫杉醇侧链连接的基因与紫杉醇生物合成呈正相关.结果表明,紫杉醇生物合成的限速步骤是催化功能官能团连接的步骤.

Secondary metabolite biosynthesis is regulated by development and inducement on and then this study explored the effects of tissue differentiation and induction on taxol synthesis in Taxus chinensis and resorted to quantitative PCR to analyze the dynamic gene expressions of the key enzymes at different stages of taxol biosynthesis. The results showed that taxol mainly distributed in bark and root-bark tissues of Taxus chinensis and appeared in the needle leaves in a very low content, and the genes of the key enzymes connected with the functional groups catalyzing taxol functions were also mainly located in the bark and root-bark tissues;Methyl jasmonate （M J） and the fungal induction factor F5 increased the taxol rates in eight and ten folds in the suspension culture of HG-1 cells of Taxus chinensis ,respectively ,and effectively induced the expression of the genes of taxol biosynthesis;The study found that the genes catalyzing the linkage of the lateral chains of taxol positively correlated with taxol biosynthesis. The results of the study revealed that the speed-limiting process of taxol biosynthesis was the process of catalyzing the connection of the functional groups.