Wu Shuangxiu, Zu Yuangang, Wu Madeline
Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, SAR, PR China.
J Biotechnol. 2003 Dec 5;106(1):33-43. doi: 10.1016/j.jbiotec.2003.07.009.
Salidroside has been identified as the most potent ingredient of the Chinese medicine herb, Rhodiola sachalinensis. Since the natural supply of this herb is rapidly decreasing, we established a compact callus aggregate (CCA) strain and culturing system for high yield salidroside production. Several callus strains induced from the explants originated from root, stem, leaf and cotyledon of R. sachalinensis were established and screened for rapid growth rate, high salidroside content and easy propagation in suspension culture condition. The CCA strain was established from a callus strain initiated from the cotyledon. The kinetics of dry weight accumulation and cellular salidroside content in various culture conditions for the strain was determined. For high salidroside production, the optimal inoculum amount was 10% and the optimal concentration for 6-benzylaminopurine and indole-3-butyric acid added in the liquid medium was 5 and 2.5 mg l-1, respectively. The acidic culture medium and a faster shaking speed favored the salidroside accumulation. The addition of 2,4-D, in the liquid MS medium and the utilization of L-tyrosol for chemical feeding enhanced salidroside production. Using a proper combination of culture condition and treatment, salidroside accumulation could reach 57.72 mg g-1 dry weight, that was 5-10-fold higher than that detected in field-grown plants. The corresponding salidroside yield was 555.13 mg l-1, a level suitable for cost effective commercial production to compensate the natural resource shortage of R. sachalinensis.
红景天苷已被确定为中药高山红景天中最有效的成分。由于这种草药的天然供应量正在迅速减少，我们建立了一种紧密愈伤组织聚集体（CCA）菌株和培养系统，用于高产红景天苷的生产。从高山红景天的根、茎、叶和子叶外植体诱导出了几种愈伤组织菌株，并对其在悬浮培养条件下的快速生长速率、高红景天苷含量和易于繁殖进行了筛选。CCA菌株是从一个由子叶起始的愈伤组织菌株建立的。测定了该菌株在各种培养条件下干重积累和细胞红景天苷含量的动力学。为了高产红景天苷，最佳接种量为10%，液体培养基中添加的6-苄基腺嘌呤和吲哚-3-丁酸的最佳浓度分别为5和2.5 mg l-1。酸性培养基和较快的振荡速度有利于红景天苷的积累。在液体MS培养基中添加2,4-D以及利用对羟基苯乙醇进行化学补料可提高红景天苷的产量。通过适当组合培养条件和处理，红景天苷积累量可达57.72 mg g-1干重，比田间种植的植物中检测到的含量高5-10倍。相应的红景天苷产量为555.13 mg l-1，这一水平适合进行具有成本效益的商业生产，以弥补高山红景天自然资源的短缺。
