目的 以马尿泡Przewalskia tangutica种子无菌苗带腋芽茎段为材料,建立了马尿泡的茎段快速繁殖体系。方法 在无菌条件下,将生长30 d左右的马尿泡种子无菌苗剪成2～3 cm长的带腋芽的茎段接种在不同的芽增殖培养基和生根培养基上,筛选出最适合芽增殖培养基和生根培养基,并利用HPLC法测定组培苗不同部位和愈伤组织的4种托烷类生物碱的量。结果 筛选出最适合芽增殖培养基为MS+6-BA 2.0 mg/L+NAA 0.5 mg/L,诱导率最高为96.67%,最适合不定芽生根培养基为MS+NAA 0.5 mg/L,根的诱导率最高,为71.67%。利用HPLC法测定试管苗不同部位4种生物碱量的结果表明,根中总生物碱量最高,高达141.25 μg/g。愈伤组织中总生物碱的量低于根、茎、叶中总生物碱的量。结论 马尿泡组培苗不同部位总生物碱的量和不同种类生物碱占总生物碱的比例不同,表明器官建成或脱分化过程可能影响马尿泡组培苗植株体内生物碱的合成和积累。

Objective To establish the stem rapid propagation culture system of Prezewalskia tangutica using aseptic seedling of P. tangutica as materials. Methods The stems from one month aseptic seedling were inoculated on Muragshige and Skoog media supplemented with different phytohormones to select the optimum medium for inducing differentiation of the adventitious buds and roots. Furthermore, the contents of four active alkaloids were measured with HPLC. Results The optimum medium was MS + 6-BA 2.0 mg/L + NAA 0.5 mg/L for inducing the propagation of the adventitious buds. The adventitious roots could be induced on MS + NAA 0.5 mg/L media. The inductivity was 71.67%. The HPLC data suggested that the content of total alkaloids is highest in root (141.25 μg/g). The content of total alkaloids in callus was lower than those in roots, stems, and leaves. Conclusion The contents of total alkaloids in different fractions of P. tangutica seedlings are different, and the proportion of different alkaloids in total alkaloids is different, which indicates that the formation and dedifferentiation of organ can influence the contents of alkaloids in P. tangutica seedlings.

国家自然科学基金资助项目(31070208);2010年度中国科学院“西部之光”人才培养计划——藏药马尿泡托烷类生物碱合成的分子调控及生物反应器的研究(Y229151211,Y129331211);中国科学院重点创新计划((KSCX2-EW-J-26)