摘要
用黄连幼叶切块，接种在含１０～２．０ｍｇ／Ｌ２，４Ｄ和０１ｍｇ／ＬＫＴ的６，７Ｖ固体培养基上，诱导形成愈伤组织。选择较松散的愈伤组织转入液体悬浮培养，获得游离细胞和细胞聚集体。通过平板培养和细胞株的筛选，选择小檗碱含量较高的细胞株。经３５次转代培养后，进行固体、液体培养，收集悬浮培养细胞进行化学提取和分离，获得黄色晶体，经ＴＬＣ、ＵＶ、ＩＲ、ＭＳ鉴定分析为小檗碱。
The callus had been induced and formed by inoculating the pieces of cut young leaves of Coptis chinensis on 6.7 V solid medium containing 1.0～2.0mg/L of 2.4 D and 0.1mg/L of kinetin.The loose callus were choosed and transfered into fluid suspension culture,free cells and cell aggregate were obtained.Cell line with higher content of berberine had been selected out by plating technique and screening.After 35times of transfering,the selected cell line was cultured on solid and in liquied medium.The cells in fluid suspension culture had been collected,extracted and isolated with chemicals,a yellow crystal could be obtained.Identificated and analysised by TLC,UV,IR,MS,it was proved that the crystal was berberine,which possessed same structure and physiological activity with that from natural plant.
出处
《生物工程学报》
CAS
CSCD
北大核心
1997年第3期284-288,共5页
Chinese Journal of Biotechnology
基金
贵州省科委资助
关键词
黄连
细胞培养
小檗碱
分离
鉴定
Coptis chinensis ,cell culture,berberine,isolation,identification
