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目的研究中国红豆杉Taxus chinensis细胞悬浮培养中蔗糖、柠檬酸三铵、水杨酸和氨基酸前体组合对细胞生长和紫杉醇(taxol)积累的影响。方法采用L9(34)设计试验,考察了第9天添加蔗糖(10、16、22 g/L)、柠檬酸三铵(0、154、1 540 mg/L),第21天添加蔗糖(20、260、g/L)和不同时间组合添加水杨酸及氨基酸前体4个因素对红豆杉细胞悬浮培养和紫杉醇合成的影响。结果当在细胞培养的第0天添加1.67 mg/L硝酸银,第9天添加10 g/L蔗糖,第9天添加1 540 mg/L柠檬酸三铵,紫杉醇达到最高,在此最优组合处理时紫杉醇质量浓度达到39.2 mg/L,相对于最差组合处理的(2.1 mg/L)时提高18.7倍。结论促进剂组合对中国红豆杉细胞悬浮培养的生长和紫杉醇的合成都有很明显的影响。
Objective To study the effects of sucrose, triammonium citrate, salicylic acid and amino acid precursors on cell growth and taxol accumulation in Taxus chinensis cell suspension cultures. Methods L9 (34) design was used to test the effects of sucrose (10,16,22 g / L), triammonium citrate (0,154,1 540 mg / L) and sucrose , 260, g / L) and different time combinations of salicylic acid and amino acid precursors on Taxus cell suspension culture and paclitaxel synthesis. Results When adding 1.67 mg / L silver nitrate on the 0th day of cell culture, 10 g / L sucrose on the 9th day, and 1 540 mg / L triammonium citrate on the 9th day, the paclitaxel reached the highest level. The optimal combination The paclitaxel concentration was 39.2 mg / L at the time of treatment, which was 18.7 times higher than that at the worst combination treatment (2.1 mg / L). Conclusion The combination of promoters has a significant effect on the growth of Taxus chinensis cell suspension culture and paclitaxel synthesis.