Gentiana scabra is an important medicinal herb that is used to treat inflammation, tuberculosis, rheumatism, cholecystitis, and hepatitis. The primary bioactive components in G. scabra are secoiridoids derived from the roots and rhizomes of the plant. In this study, we sought to establish a means of reliably producing these secoiridoids through G. scabra in vitro adventitious root culture. To that end, we studied the effect of phytohormones, light condition, explant type, and medium composition on adventitious root induction and secoiridoid production. Maximal induction frequency (100%) was ultimately achieved when using leaf explants cultured on Murashige and Skoog (MS) medium containing 3.0 mg L⁻¹ NAA and 0.25 mg L⁻¹ TDZ with a 16/8-h photoperiod, yielding 53.20 ± 3.15 roots per explant. We explored root growth curves over an 8-wk period in MS liquid medium prepared as above, and observed maximal fresh weight (FW) and dry weight (DW) values of 26.94 ± 0.38 g per flask and 1.79 ± 0.032 g per flask, respectively, in 4-wk-old cultures. The accumulation of secoiridoids (gentiopicroside, swertiamarin, and sweroside) in these adventitious root cultures was assessed via high-performance liquid chromatography (HPLC), revealing maximal gentiopicroside (25.59 ± 0.65 mg g⁻¹ DW), swertiamarin (1.61 ± 0.04 mg g⁻¹ DW), and sweroside (4.42 ± 0.11 mg g⁻¹ DW) levels after 4 wk. Together, our results reveal that G. scabra adventitious root cultures represent a viable approach to the in vitro production of secoiridoids. This is the first study to our knowledge to have explored the isolation of secoiridoids following the in vitro growth of G. scabra adventitious roots. These data may represent an important step toward the development of large-scale adventitious root cultures for commercial secoiridoids production.

龙胆草是一种重要的药材，用于治疗炎症、肺结核、风湿病、胆囊炎和肝炎。G. scabra中的主要生物活性成分是来自植物根和根茎的二环萜类化合物。在这项研究中，我们试图建立一种通过G. scabra 体外不定根培养可靠生产这些类环烯醚萜类化合物的方法。为此，我们研究了植物激素、光照条件、外植体类型和培养基成分对不定根诱导和类环烯醚萜类生成的影响。当使用在含有 3.0 mg L ^-1 NAA 和 0.25 mg L ^{-1 的}Murashige 和 Skoog (MS) 培养基上培养的叶外植体时，最终实现了最大诱导频率 (100%)TDZ 的光周期为 16/8 小时，每个外植体产生 53.20 ± 3.15 个根。我们在如上制备的 MS 液体培养基中探索了 8 周内的根生长曲线，并观察到最大鲜重 (FW) 和干重 (DW) 值分别为每瓶 26.94 ± 0.38 克和每瓶 1.79 ± 0.032 克，在 4 周前的文化中。通过高效液相色谱法 (HPLC) 评估了这些不定根培养物中的秒环烯醚萜苷（龙胆苦苷、芥末苷和 sweroside）的积累，揭示了最大的龙胆苦苷（25.59 ± 0.65 mg g ^-1 DW）、芥末苷（1.61 mg ± 0.04 mg ± 0.04 mg）^-1 DW) 和4 周后的苦参苷 (4.42 ± 0.11 mg g ^-1 DW) 水平。总之，我们的结果表明G. scabra不定根培养代表了体外生产二环萜类化合物的可行方法。据我们所知，这是第一项探索G. scabra不定根体外生长后分离类环烯醚萜类的研究。这些数据可能代表了开发用于商业化环萜类化合物生产的大规模不定根培养的重要一步。