Fluridone-Mediated Strigolactone Inhibition Enables Complete Micropropagation of Echinacea purpurea: From Shoot Multiplication to Field Establishment

Background Echinacea purpurea represents a high-value medicinal crop with global market demand exceeding $338 million annually. However, micropropagation faces significant limitations due to strigolactone-mediated apical dominance, while complete protocols encompassing rooting and acclimatization phases remain underdeveloped for commercial applications. Methods We systematically optimized fluridone concentrations (0–10 µM) for shoot multiplication through strigolactone biosynthesis inhibition, developed factorial IBA×NAA rooting protocols, and implemented a three-stage acclimatization system with controlled environmental transitions. Endogenous strigolactone quantification via LC-MS/MS validated the mechanistic basis. Results Optimal fluridone treatment (3 µM) achieved 9.7 ± 0.6 shoots per explant, representing a 273% increase over controls (P < 0.001, η² = 0.647). Root induction using 2.5 µM IBA combined with 0.5 µM NAA yielded 8.3 ± 0.4 roots per shoot with 95.3% success rate and superior architecture scores (4.6 ± 0.3). The integrated three-stage acclimatization protocol achieved 89.3% field survival with robust plant quality parameters. Complete protocol success rate reached 76.2% from explant to field-ready plants in 16 weeks. Conclusions This strigolactone-targeted approach enables efficient commercial-scale E. purpurea production, addressing critical bottlenecks in medicinal plant biotechnology while providing mechanistic insights into hormonal regulation in tissue culture systems. The protocol supports sustainable production of 106,000 plants annually from 1,000 explants.