粉叶小檗愈伤组织单细胞克隆

以粉叶小檗愈伤组织为材料 ,用B5液体培养基进行悬浮培养建立悬浮细胞系。经 3～ 4次继代培养即可得到悬浮的单细胞。悬浮细胞通过细胞平板克隆 (一般B5培养基平板克隆 ,优化培养基平板克隆和条件培养基平板克隆 ) ,经 5代连续继代培养观察和薄板层析 -分光光度法分析 ,发现用优化培养基进行平板克隆植板率最高 ,且克隆最易成功 ,并且还筛选到一株小檗碱产率高且稳定的克隆CV 5 7,其平均生长速率为 14 .4 12mg .Fw/L .d ,为原始株系的 1.91倍 ,平均小檗碱含量为 2 .17%干重 ,是原始株系的 2 .2 6倍。

A Single Cell Clones From Calli of Berberis Pruinosa

The calli of Berberis pruinosa were used as material and were transferred to fluid medium B5 , which was suitable for suspension culture of calli cell , and in the result the cell suspension line was established .Suspension single cells were obtained after successive subculturing of 3 or 4 generations.With cell plate clone (normal plate culture, superiority culture and condition culture)of suspension cells, it was found that the plating efficiency was the highest in superiority media. Furthermore, a more stable high yield Berberine clone line CV 57 had been selected according to successive subculturing of 5 generation and analysed by TLC Spectrophotometry. The average growth rate of clone line CV 57 was 14.412mg.Fw/L.d and was 1.91 folds higher than to the original strain. Its average content of Berberine was 2.17%(DW) and was 2.26 folds higher than that of the original strain.