An efficient and scalable in vitro propagation protocol of Atractylodes macrocephala Koidz.: A key traditional medicinal plant within Food-Medicine Continuum

Atractylodes macrocephala Koidz., an important medicinal species within the food–medicine continuum, is widely utilized across Korea, China, and Japan. However, its traditional seed-based cultivation is increasingly unsustainable due to low germination rates, slow multiplication, and short seed viability, necessitating alternative propagation strategies to meet growing demand. This study developed an efficient and scalable in vitro propagation protocol using shoot tip explants. The effects of culture media, cytokinins for shoot proliferation, and auxins for root induction were assessed, alongside physiological, biochemical, and genetic comparisons between in vitro-regenerated and maternal plants. Shoot tips cultured on Murashige and Skoog (MS) medium supplemented with 1.5 mg/L benzyl aminopurine (BAP) yielded the highest shoot proliferation, producing 17.5 ± 1.21 vigorous shoots per explant. Optimal rooting was achieved on MS medium with 1.5 mg/L indole-3-acetic acid (IAA), generating 21.2 ± 1.32 roots with an average root length of 8.76 ± 0.85 cm and a surface area of 252.20 ± 4.92. Acclimatization produced a 93.33% survival rate, indicating strong adaptation of regenerated plantlets to ex vitro conditions. Comparative assessments showed that regenerated plants displayed similar chlorophyll content, photosynthetic efficiency, genetic stability (flow cytometry), and biochemical characteristics (FT-NIR) to maternal plants, confirming the fidelity and quality of in vitro-derived plants. Overall, this cost-effective and reproducible micropropagation protocol enables rapid mass production of uniform, disease-free A. macrocephala plantlets. The platform offers significant potential for conservation, commercial production, and long-term germplasm preservation of this valuable food-medicinal plant.