粉叶小檗愈伤组织单细胞克隆  

A single cell clones from calli of Berberis pruinosa

作　　者：查宏波 李启任 王东 陈敏 

机构地区：[1]云南省烟草昭通市公司烟草科学研究所,云南昭通657000

出　　处：《生物技术通讯》2003年第4期291-293,共3页Letters in Biotechnology

摘　　要：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-57,其平均生长速率为每天14.412mg/L,为原始株系的1.91倍,平均小檗碱含量为干重的2.17%,是原始株系的2.26倍。The calli of Berberis pruinosa were used as material and were transferred to fluid medium B 5 ,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 of3or4generations.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-57had been selected according to successive subculturing of5generation and analysed by TLC-Spectrophotometry.The average growth rate of clone line CV-57was14.412mg /(L·d)and was1.91folds higher than to the original strain.Its average content of berberine was2.17%(DW)and was2.26folds higher than that of the original strain.

关 键 词：粉叶小檗 愈伤组织 单细胞克隆 克隆系 平板克隆 小檗碱 

分 类 号：Q943[生物学—植物学] Q813.11