目的为扩大和提高肉苁蓉Cistanchedeserticola资源的利用效率。方法以肉苁蓉肉质茎的不同组织部位作为外植体,采用正交实验方法,筛选诱导愈伤组织产生的不同培养基和培养条件,并继代培养。应用HPLC方法对愈伤组织培养物中松果菊苷和洋丁香苷(毛蕊花糖苷)的量进行测定。结果肉苁蓉肉质茎的维管组织部分是诱导愈伤组织的最适外植体,鳞片叶次之,髓组织部分诱导效果较差。在暗培养、25~27℃条件下以B5为基本培养基附加6-BA(0.5~2mg/L)与IAA(0.5~1.5mg/L)诱导愈伤组织效果最佳;在半光照(光培养10h/d,暗培养14h/d)条件下,愈伤组织生长正常,最佳继代时间为25~30d。愈伤组织培养物中松果菊苷和洋丁香苷(毛蕊花糖苷)达4.37%。结论筛选出了适宜的肉苁蓉愈伤组织培养方法,且培养物中主要药用有效成分松果菊苷和洋丁香苷(毛蕊花糖苷)的量达到并超过了《中国药典》要求(0.3%)的标准。

Objective In order to explore and improve the utilization efficiency of Cistanche deserticola resources. Methods The different tissue parts of fleshy stem of C. deserticola were tested as the explants and cultured on various culture media and condition for callus induction and subculture by orthogonal test. The content of acteoside in callus was determined by HPLC. Results The vascular tissue parts of fleshy stem of C. deserticola was optimum explants of callus induction, the following was scale leaf, while pith tissue was relatively bad. Under 25 ℃— 27 ℃ conditions, B5 as basic media and adding 6-BA (0.5—2 mg/L) and IAA (0.5—1.5 mg/L) was able to achieve the best effect of callus induction by growing in the dark. Under given half light (the light culture was 10 h/d, the dark culture was 14 h/d) conditions, the growth of callus was normal, the optimum time of subculture was 25—30 d. The content of echinacoside and acteoside in callus was 4.37%. Conclusion The optimized culture method of callus of C. deserticola is obtained and the content of the primary medical constituent in callus has beed up to or even over the standard (0.3%) in Pharmacopoeia of China.

国家自然科学基金项目(30260008)；国家科技部攻关西部专项(2002BA901A32)；宁夏自然科学基金项目(NZ0522)