藏红花细胞悬浮培养体系的建立及优化  

Establishment and Optimization of Crocus sativus L. Cell Suspension Culture System

作　　者：陈书安[1,2] 王晓东[1] 袁晓凡[1] 赵兵[1] 王玉春[1] 

机构地区：[1]中国科学院过程工程研究所生化工程国家重点实验室,北京100190 [2]科技部中国生物技术发展中心,北京100036

出　　处：《生物技术通报》2010年第7期157-160,共4页Biotechnology Bulletin

基　　金：国家"863"计划(2002AA241121)

摘　　要：基于诱导的藏红花细胞系,通过摇瓶法,优化了其液体培养基、接种量和种龄等培养条件,以建立藏红花细胞悬浮培养体系。结果表明,将生长在固体培养基上的藏红花愈伤组织接种在MS液体培养基(添加了2mg/L2,4-D,1mg/L6-BA和300mg/LCH)中,于(22±0.3)℃,120r/min的摇床上,暗培养30d,便可获得藏红花的悬浮细胞系。经优化其培养基、接种量和种龄,将种龄为20d的细胞系,按照5%接种量接种在液体B5培养基(添加了2mg/LNAA,1mg/L6-BA和300mg/LCH)中,于(22±0.3)℃,120r/min的摇床上,培养36d,细胞生物量(13.4g/L)和藏红花素产量(0.91g/L)均达到最高。本研究建立的藏红花细胞悬浮培养体系为其生物反应器放大培养奠定了基础。Based on the screened callus line,the liquid medium,inoculation size and cell age were optimized by shake-flask culture to establish the cell suspension culture system of Crocus sativus L. cell. When callus was cultured in MS liquid medium supplemented with 300 mg/L CH, 1 mg/L 6-BA and 2.0 mg/L 2,4-D on shaking bed at（22 ± 0.3 ） ℃ for 30 d, its suspension cell could be obtained. The maximum biomass（ 13.4 g/L） and crocin production （ 0.91 g/L） reached when 20 d age cells were cultured in B5 liquid medium supplemented with 300 mg/L CH ,2 mg/L NAA and 1 mg/L 6-BA by 5% inoculation size. The cell suspension culture system established in this research could provide references for the scale-up culture of Crocus sativus L. cell by bioreactor.

关 键 词：藏红花 悬浮 培养体系 摇瓶 生物反应器 

分 类 号：S567.219[农业科学—中草药栽培]