Ligusticum chuanxiong Hort. is a renowned genuine medicinal material from Sichuan with significant medicinal value. However, the lack of an efficient tissue culture system and quality deterioration in conventional cultivation—caused by pests, diseases, environmental pollution, and continuous cropping obstacles—have hindered the sustainable utilization and industrial development of its superior germplasm resources. This study successfully developed an efficient regeneration system via suspension cell cultures of L. chuanxiong. Callus induction from petioles on MS medium supplemented with 1.5 mg/L 2,4-D, 1.5 mg/L 6-BA, and 30 g/L sucrose achieved an induction rate of 86.15%. The callus was then transferred to liquid MS medium containing 1 mg/L 2,4-D and 1 mg/L 6-BA at an inoculation density of 40 g/L, yielding embryogenic suspension cells capable of sustained growth and division. When cultured on solid MS medium with 1 mg/L 6-BA and 0.50 mg/L NAA, the suspension cell aggregates achieved a shoot regeneration rate of 93.64%. The highest rooting rate (76.67%) was obtained on MS medium containing 1 mg/L IBA and 0.5 mg/L NAA. Rooted plantlets transplanted into pots with a peat: perlite ratio (2:1 v/v) showed a survival rate of 90.91% after 3–4 weeks. Genetic fidelity analysis using four ISSR primers revealed no somaclonal variation. Treatment with 100 μmol/L salicylic acid increased senkyunolide A and senkyunolide I contents by 10.10-fold and 25.54-fold, respectively, while 150 μmol/L salicylic acid enhanced butylidenephthalide, ligustilide, caffeic acid, and biomass by 10.29-fold, 3.73-fold, 2.84-fold, and 1.27-fold, respectively. This study accomplished rapid, large-scale plant regeneration through L. chuanxiong suspension cells, offering essential support for industrial propagation, an ideal cell source for genetic transformation, and key solutions to germplasm degradation.
Chen et al. (Tue,) studied this question.