灵芝诱导子对南方红豆杉悬浮培养细胞产紫杉醇的影响

Effect of Ganoderma lucidum Elicitor on Paclitaxel Production by Taxus wallichiana var.mairei Suspension Culture Cells

为提高红豆杉悬浮细胞紫杉醇产量,采用制备的灵芝(Ganoderma lucidum)诱导子处理南方红豆杉(Taxus wallichiana var.mairei)悬浮细胞,通过分析细胞生长量、紫杉醇总产量,研究灵芝诱导子对红豆杉悬浮细胞的影响。结果表明,灵芝诱导子添加质量浓度为100µg/mL时,红豆杉悬浮细胞生长指数和紫杉醇总产量显著高于其他质量浓度(P<0.05),该质量浓度为灵芝诱导子的最佳添加质量浓度;灵芝诱导子在细胞生长的第8天加入时,第21天的细胞生长指数和紫杉醇总产量显著高于其他添加时间(P<0.05),细胞生长的第8天为灵芝诱导子的最佳添加时间;灵芝诱导子持续作用6 d时,细胞生长指数和紫杉醇总产量显著高于其他持续作用时间(P<0.05),该持续作用时间为灵芝诱导子的最佳持续作用时间。在悬浮细胞培养的第8天加入100µg/mL灵芝诱导子培养至第21天时,细胞干质量和紫杉醇总产量达到峰值,分别为对照(等量蒸馏水处理)的221.13%、569.69%;细胞膜表面黏膜和球状物质增加;苯丙氨酸解氨酶活性和总酚含量提高,细胞活力和多酚氧化酶活性降低。

To improve the production of paclitaxel in suspension cells of Taxus wallichiana var.mairei,Ganoderma lucidum inducers were prepared to treat the suspension cells of Taxus wallichiana var.mairei.The effects of Ganoderma lucidum inducers on the suspension cells of Taxus wallichiana var.mairei were studied by analyzing cell growth and total production of paclitaxel.The results showed that when the concentration of Ganoderma lucidum elicitor added was 100µg/mL,the growth index and total production of paclitaxel of Taxus wallichiana var.mairei suspension cells were significantly higher than other concentrations(P<0.05),so 100µg/mL was the optimal concentration for adding Ganoderma lucidum elicitors;When the Ganoderma lucidum elicitor was added on the 8th day of cell growth,the cell growth index and total production of paclitaxel on the 21st day were significantly higher than other addition time(P<0.05),so the 8th day of cell growth was the optimal addition time for the Ganoderma lucidum elicitor;When the Ganoderma lucidum elicitor continuously treated the suspension cells for 6 days,the growth index of suspension cells and total production of paclitaxel were significantly higher than other sustained treatment time(P<0.05),so 6 days were the optimal duration of the Ganoderma lucidum elicitor.When 100µg/mL of Ganoderma lucidum elicitor was added on the 8th day of suspension cell culture to the 21st day,the dry weight of cells and the total production of paclitaxel reached the peak,which were 221.13%and 569.69%of the control(adding the same amount of distilled water),respectively;Membrane surface mucosa and globular substances increased;Phenylalanine ammoniase activity and total phenol content were increased,while cell viability and polyphenol oxidase activity were decreased.