Abstract
A rapid and improved micropropagation protocol was developed for Curcuma aromatica, a threatened aromatic medicinal plant, using rhizome sprout as the explant. Stepwise optimization of different plant growth regulators, carbon sources, and basal media was adopted to establish an efficient micropropagation protocol. When cytokinins, such as benzyl amino purine (BAP) or 6-(alpha,alpha-dimethylallylamino)-purine (2iP), were used either singly or in combination with naphthalene acetic acid (NAA) for shoot induction and multiplication, a single use of BAP was the most effective. As a carbon source, 3% (w/v) sucrose exhibits the greatest promotive effect on shoot initiation and proliferation compared with other carbon sources used. Among the basal media, full strength Murashige and Skoog (MS) media produced the best results, compared to other media studied. By using the most effective treatment from each category, an average of 13.2 shoots/per explant were produced after 6 weeks of culture. Moreover, 85% survival was achieved when rooted explants acclimatized ex vitro using a mixture of sterile sand, soil, and farmyard manure (1: 1: 1). In addition, antimicrobial activities of rhizome extracts were evaluated. Petroleum ether and chloroform extracts of field-grown rhizome showed potential antimicrobial properties against several human pathogenic bacteria including Bacillus subtilis, Staphylococcus aureus, Pseudomonas aeruginosa, Shigella sonnei, and Shigella dysenteriae, with a minimum inhibitory concentration (MIC) ranging from 0.03 to 0.5 mg/mL. Thus, the optimized micropropagation protocol may offer large-scale production of plantlets to meet industrial demand for the rhizome. Moreover, our results suggest the rhizome extract of C. aromatica is a promising antimicrobial agent.