In Vitro Production of Plantlets and Microrhizomes, Genetic Fidelity Assessment, and Metabolic Profiling of Boesenbergia rotunda (L.) Mansf.

Fingerroot (Boesenbergia rotunda (L.) Mansf.) is valued for its therapeutic benefits, both in Thailand and internationally. This study optimized in vitro propagation and induced microrhizomes (MRZ) to produce cleaned plantlets to support organic farming using disease-free plantlets, which is crucial for preventing and eradicating diseased plantlets, reducing the use of chemicals, and alternative approaches to enhancing phytochemical diversity. Shoots cultured on ½-strength MS medium with 1 mg L−1 of 6-benzylaminopurine (BAP) showed the highest shoot formation (69%) and shoot multiplication (3.45 ± 0.29 shoots per explant). Plantlets acclimatized in peat moss or a peat moss–coconut coir (1:1) mixture achieved a 100% survival rate. Genetic fidelity was confirmed using SSR markers, showing genetic consistency with the mother plant. The MRZ formation was the highest (98.33%) under white LED light with 30 g L−1 of sucrose. Nuclear magnetic resonance (NMR) analysis in MRZ revealed aspartate, a precursor to pinocembrin and pinostrobin. Additionally, nine unique metabolites not previously identified in fingerroot were detected in the MRZ, suggesting some potential in novel therapeutic applications. These findings support the development of efficient micropropagation methods and highlight MRZ as a source of diverse bioactive compounds, contributing to the medicinal value of B. rotunda in sustainable and large-scale production.