几种理化因子对青钱柳悬浮细胞生长的影响

Effects of Several Factors on the Growth of Cyclocarya paliurus Suspension Cells

为了利用细胞悬浮培养技术生产青钱柳细胞及其代谢物质以解决青钱柳资源稀缺问题,采用单因素和正交试验优化青钱柳细胞悬浮培养条件。结果表明,在供试的WPM、MS、1/2MS、B5、N6、DKW、White7种培养基中,WPM、MS、B5均有利于悬浮细胞的生长;有利于悬浮细胞生长的碳源是果糖和蔗糖;2,4-D是影响青钱柳悬浮细胞生长的关键因子,对细胞生长影响极显著,最佳激素组合为0.4mg/L 2,4-D+0.4mg/LNAA+0.4mg/L KT+0.4mg/L 6-BA;通过L9(34)正交试验得出几种物理因子对悬浮细胞生长的影响效应排序为接种量>装液量>转速>pH,最佳条件组合为接种量80g/L、装液量40%、摇床转速115r/min、pH 5.0。

In order to solve the scarcity of Cyclocarya paliurus resources, the cell suspension culture technology was used to produce C. paliurus cells and its metabolites. The optimal growing conditions of suspension cells were investigated,by single factor and orthogonal experiment. The results showed as follows： among the seven basic medium of WPM, MS, 1/2MS, B5, N6, DKW and White tested, WPM, MS and B5 were favorable for the growth of suspension cells. Sucrose and fructose were favorable for the growth of suspension cells. 2,4-D was the key factor for the growth of suspension cells. Suspension cells grew well in MS medium supplemented with 0.4 mg/L 2,4-D, 0.4 mg/L NAA, 0.4 mg/L KT and 0.4 mg/L 6-BA. Experimental results indicated that the impact order of four factors on the growth was listed as follows： inoculum densities〉liquid volume 〉 rotation speed 〉initial pH, the optimal culture condition combination was 8% of Inoeulum densities, 40% of liquid medium volume, 115 r/min of rotation speed and initial pH 5.0.