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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 ,较早期的代谢中间产物对紫杉醇合成影响较弱
Using Taxus chinensis var. Meirei suspension cells as materials, the synthesis pathway diversity and metabolite intermediates were studied using 3-hydroxy-3-methylglutarate reductase (HMGR) inhibitors in the paclitaxel synthesis pathway. Effects of geraniol, geraniol, terpene, mevalonate and elicitors (arachidonic acid, methyl jasmonate) on cell growth and paclitaxel content in culture to explore taxol biosynthesis The possibility of artificial regulation. Results: From the growth curve of cultured cells, cell dry weight and paclitaxel content reached the highest level at the 30th day of culture. The inhibitory effect of the HMGR inhibitor compactin on taxol biosynthesis was not significant. At the beginning of the culture, the calf yak was added. Alcohol (5mg/L) can significantly increase the content of paclitaxel, geraniol (10mg/L), mevalonate (100mg/L) can also promote the synthesis of paclitaxel, while terpenes (10mg/L). No effect; addition of arachidonic acid (3.2 μmol/L) and methyl jasmonate (10 μmol/L) increased the content of paclitaxel by 1 and 10 times, respectively. Conclusion : Taxol biosynthesis may have other pathways in addition to the classical acetate/mevalonate pathway in Taxus chinensis culture cells. The addition of appropriate metabolic intermediate elicitors can increase paclitaxel content in plant culture cells, but due to the complex biosynthetic steps, the earlier metabolic intermediates have a weaker effect on paclitaxel synthesis.