High-Frequency Somatic Embryogenesis and Molecular Validation of Genetic Fidelity in Kaempferia marginata Carey ex Roscoe: An Endangered Medicinal Plant

Abstract

Kaempferia marginata Carey ex Roscoe is a valuable medicinal plant currently threatened by over-exploitation and habitat loss. We established an efficient in vitro micropropagation protocol via somatic embryogenesis. Rhizome buds were cultured on Murashige and Skoog (MS) medium supplemented with varying concentrations of auxins and cytokinins. The highest frequency of callus induction (87.00 ± 0.10%)—producing friable, embryogenic callus—and subsequent shoot regeneration (7.5 ± 0.16 shoots/explant) was achieved with 2.0 mgL⁻¹ α-naphthalene acetic acid (NAA). Histological analysis confirmed the ontogeny of somatic embryos progressing from globular to torpedo stages. For multiplication, a combination of 4.0 mgL⁻¹ 6-benzylaminopurine (BAP) and 1.0 mg L⁻¹ NAA yielded the maximum number of shoots (16.35 ± 0.22) within 4 weeks. Genetic stability was assessed using 25 RAPD and 11 ISSR primers, which generated 84 and 76 reproducible bands, respectively. All profiles were monomorphic across regenerants and the mother plant, confirming true-to-type propagation suitable for large-scale conservation and commercial utilization.