Breaking the bottleneck in Dendrobium officinale plantlets production: A novel “multiplication-rooting integration” system with dual regeneration pathways

The large-scale propagation of Dendrobium officinale Kimura et Migo, a precious medicinal orchid, is hindered by long seedling cycles, high genetic heterogeneity, and costly conventional micropropagation. This study aimed to develop an efficient integrated in vitro regeneration system that combines shoot multiplication and rooting in a single medium. Through orthogonal optimization of basal medium and plant growth regulators, an optimal formulation was established. This system achieved a multiplication coefficient of 10.21, 100% rooting within 90 days, and an additional protocorm-like body pathway with a multiplication coefficient of 20.34. Plantlets showed > 95% survival after transplanting. The system shortens the production cycle, reduces costs, and ensures genetic uniformity, providing a commercially viable approach for large-scale production and sustainable industrial utilization of D. officinale. • Integrated system enables simultaneous shoot proliferation and rooting in a single medium. • Production cycle shortened from 12 months to 70–90 days, with a proliferation coefficient of 10.21. • Genetic uniformity is ensured via nodal segment direct organogenesis, avoiding trait segregation. • Dual regeneration pathways provide flexibility for high-fidelity, high-yield propagation. • Scalable, cost-effective system has industrial adoption, with 25 million seedlings produced.