肉苁蓉细胞悬浮培养体系的建立

Establishment of Cistanche deserticola cell lines in suspension culture

目的研究由肉苁蓉愈伤组织建立细胞悬浮培养体系的培养条件。方法在固体培养基上添加不同比例的生长激素,考察生长激素、细胞种龄和接种量对细胞生长和次生代谢产物苯乙醇糖苷(PeG)量的影响。结果在添加1mg/L2,4-D的B5固体培养基上形成的愈伤组织松散易碎,适于进行悬浮培养。细胞种龄为25d和接种量为3%～4%时细胞悬浮培养的结果最好,PeG产量达到最大,为0.82g/L。细胞在悬浮培养时聚集体由大变小,直径小于0.5mm的细胞聚集体由接种时的5%逐渐增加至第20天时的81%,其PeG量为10.1%。结论通过摇瓶培养建立了较好的细胞悬浮培养体系。

Objective To investigate the culture conditions for the establishment of suspension cell lines from Cistanche deserticola callus. Methods Adding different kinds and ratios of plant growth regulator to solid B5 medium, the effects of cell age, inoculation size, and plant growth regulator on the cell growth and the second metabolite accumulation, phenylethanoid glycosides （PEG） were studied. Results Callus formedon solidified B5 medium supplemented with 1 mg/L 2,4-D was spongy and suitable for suspension culture. The production of PeG reached the highest of 0.82 g/L when cell suspended culture with the cell age of 25 d and inoculation size of 3%-4%. In suspension culture, the ratio of cell aggregation smaller than 0. 5 mm changed from 5% at the start to 81% at the day 20. It＇s PeG content reached 10. 1%. Conclusion Optimal cell suspension lines are established after flask culture.