目的 建立蒺藜Tribulus terrestris愈伤组织培养体系,并考察种子、全草及愈伤组织中总黄酮醇苷量。方法 采用正交试验研究不同外植体、培养基、光照条件、激素种类及其配比等对蒺藜愈伤组织诱导和增殖的影响,确定蒺藜愈伤组织培养体系,采用HPLC测定其种子、全草及愈伤组织中总黄酮醇苷量。结果 以蒺藜茎段为外植体,诱导愈伤组织的适宜培养基为MS+2,4-D 0.5 mg/L+6-BA 0.1 mg/L,25℃全天恒温暗培养;最佳增殖培养基为MS+2,4-D 1.0 mg/L+6-BA 0.2mg/L+ZT 1.0 mg/L,12 h/d光培养,pH 6.2,继代周期为35 d。种子、全草及愈伤组织中总黄酮醇苷量分别为1.46%、2.01%、5.22%。结论 愈伤组织质地疏松,适合建立稳定高效的组织培养体系,其有效成分总黄酮醇苷量高于种子和全草。

Objective To establish the callus culture system of Tribulus terrestris and investigate the contents of the total flavonoid glycosides in its seeds, whole herb, and callus. Methods By using the orthogonal test design, different explants, the effects of culture medium, light condition, suitable hormones, and their ratio on callus induction and proliferation were studied. Callus culture system of T. terrestris was determined and the total flavonoid glycosides of its seeds, whole herb, and callus were detected. Results The stems of T. terrestris were used as the experimental materials. The optimal medium for the callus induction was MS+2,4-D 0.5 mg/L+6-BA 0.1 mg/L, with temperature of 25℃ and darkness of 24 h/d. The optimal medium for callus differentiation was MS+2,4-D 1.0 mg/L+6-BA 0.2 mg/L+ZT 1.0 mg/L, with light of 12 h/d, pH of 6.2, and subculture cycle of 35 d. The total flavonoid glycosides in the seeds, whole herb, and callus of T. terrestris were 1.46%, 2.01%, and 5.22%. Conclusion Callus induced by tissue culture technique are soft, luster, and transparent, which are suitable for establishing the stable and efficient callus culture system of T. terrestris. The total flavonoid glycosides in callus are more than those in the seeds and whole herb.

吉林省科技发展计划项目(20120923)