南方红豆杉细胞悬浮培养体系优化及动力学研究  

Study on Optimization of Cell Suspension Culture System and Kinetics of Taxus chinensis var. mairer

作　　者：徐志荣 王婷[1] 娄佳兰 魏赛金[1] XU Zhi-rong;WANG Ting;LOU Jia-lan;WEI Sai-jin(Jiangxi Agricultural University /Jiangxi Agricultural Microbial Resource Development and Utilization Engineering Lab, Nanchang 330045, Jiangxi, China)

机构地区：[1]江西农业大学/江西省农业微生物资源开发与利用工程实验室,江西南昌330045

出　　处：《林业科学研究》2019年第1期8-14,共7页Forest Research

基　　金：江西省科技计划项目(20133BBG70090);江西农业大学大学生创新创业训练项目(201710410067)

摘　　要：[目的]南方红豆杉细胞悬浮培养体系优化及动力学模型构建。[方法]以南方红豆杉1年生枝条诱导产生的愈伤组织为材料,在优化愈伤组织接种量、培养基初始pH、植物激素类型及配比的基础上,分析南方红豆杉悬浮细胞生长和紫杉醇积累的动力学关系及主要营养成分的变化。[结果]适合南方红豆杉悬浮培养的培养基为:B5+0.4 mg·L^(-1) 2,4-D+0.3 mg·L^(-1) NAA+1.2 mg·L^(-1) 6-KT+30 g·L^(-1)蔗糖,培养基最佳初始pH为5.8,最佳接种量为0.09 g·mL^(-1)。在1个培养周期内,培养液中的糖、磷元素,铵态氮、硝态氮在细胞培养的第18天基本被消耗完全或保持不变;动力学拟合表明,南方红豆杉细胞悬浮培养生长符合Logistic生长模型,最大比生长速率为0.143 65 d^(-1),底物消耗和紫杉醇合成可用Luedeking-Piret模型描述,悬浮细胞生长与紫杉醇积累属于部分生长偶联型。[结论]南方红豆杉细胞悬浮培养体系优化后,通过模型的回归拟合,获得了反映南方红豆杉细胞培养的动力学参数,试验所构建的发酵动力学模型在一定程度上揭示了南方红豆杉细胞产紫杉醇代谢过程的特征。[Objective] To optimizing the cell suspension culture system and building the kinetics model of Taxus chinensis var. mairer .[Method] The effects of basic callus inoculum size, initial pH value, phytohormone types and concentration on T. chinensis var. mairer cell growth was investigated using the callus induced from annual branches of T. chinensis var. mairer . Based on this, the kinetic relationship between T. chinensis var. mairer cell growth and paclitaxel production and the nutrient consumption were also analyzed.[Result] The optimal media for T. chinensis var. mairer cell suspension culture is B5 medium supplemented with 0.4 mg·L^-1 2,4-D, 0.3 mg·L^-1 NAA, 1.2 mg·L^-1 6-KT and sucrose 30 g·L^-1 . The optimum callus inoculum size is 0.09 g·mL^-1 and the optimum initial pH of medium is 5.8. During the process, the sugar, phosphor-N and nitrate-N in the culture medium are nearly used up or remain unchanged on the 18th day. The growth law of T. chinensis var. mairer is consistent with Logistic growth model and the maximum specific growth rate is 0.143 65 d^-1 . Substrate consumption and paclitaxel production by Luedeking-Piret model are described. The growth of suspension cell and the accumulation of paclitaxel belong to partial growth coupling type.[Conclusion] The cell suspension culture system is optimized. Through regression model, the kinetic parameters are obtained to reflect the cell culture of T. chinensis var. mairer . The fermentation kinetics model constructed in this experiment reveals the characteristics of paclitaxel production in Taxus chinensis cells to a certain extent.

关 键 词：南方红豆杉 悬浮细胞 紫杉醇 优化 动力学模型 

分 类 号：S718.46[农业科学—林学]