切花菊悬浮细胞系的建立及生长规律

Establishment of Cell Suspension Culture and Growth Regularity of Dendrathema morifolium

以切花菊‘粉贵人’(Dendrathema morifolium‘Fenguiren’)淡黄色、质地疏松、增殖速度快的愈伤组织为起始材料,接种在MS+2,4-D0.5 mg/L+6-BA0.2 mg/L+30 g/L蔗糖的液体培养基上,在23℃黑暗条件下振荡(100 r/min)培养30～40 d,建立分散性好、均匀、生长迅速的悬浮细胞系。在悬浮细胞系培养过程中研究了悬浮细胞的生长规律,结果表明:在40 mL培养液中,接种细胞起始数量最适为1.6×103～4.8×103个;pH值范围在5.29～5.68时,悬浮细胞增殖率最高,继代时保留1/3体积的旧培养液有利于其细胞增殖。

Dendrathema morifolium ＇ Fenguiren＇ of pale yellow callus with loose structure and high proliferation rate was selected as material for cell suspension culture. It was cultured in modified liquid MS medium MS＋2,4-D0.5 mg/L＋6-BA0.2 mg/L＋30 g/L, and shook at 100 r/min under dark condition at 23 ℃. About 50-40 days later, stably growing cell suspension cultures were established. The growth pattern of the cultures was investigated. The results showed there are 1.6×10^3 -4.8×10^3 initial inoculation cells in the liquid medium of 40 mL. In 5.29-5.68 of pH, suspension cell has the highest rate of cell proliferation. 1/3 of old liquid medium left is fit for cell proliferation during subculture.