Research Perspective on Biological Science Vol. 11

Kaempferia marginata Carey ex Roscoe (Zingiberaceae) is a small perennial herb that belongs to the Zingiberaceae family. The present study aims to standardise a highly efficient and reproducible in vitro micropropagation protocol via somatic embryogenesis for Kaempferia marginata, an endangered and medicinally significant herb, and to concurrently validate the genetic fidelity of the regenerated plantlets. Because traditional propagation via rhizomes is extremely slow and susceptible to soil-borne pathogens, somatic embryogenesis was selected to provide a high-frequency, scalable alternative. Sprouting rhizome buds were utilised as explants and cultured on Murashige and Skoog (MS) basal medium. To evaluate callus induction and subsequent shoot multiplication, the media were supplemented with various concentrations of plant growth regulators (PGRs), including α-naphthalene acetic acid (NAA), 2,4-Dichlorophenoxyacetic acid (2,4-D), indole acetic acid (IAA), 6-benzylaminopurine (BAP), and Kinetin. Furthermore, to assess the genetic uniformity of the in vitro regenerants against the mother plant, genomic DNA was extracted, and molecular profiling was conducted using Random Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeat (ISSR) markers. The highest frequency of embryogenic callus induction (87.00 ± 0.10%) was achieved on MS medium fortified with 2.0 mg L⁻¹ NAA. Following the developmental stages of somatic embryos (globular to torpedo), transfer to regeneration media revealed that a synergistic combination of 4.0 mg L⁻¹ BAP and 1.0 mg L⁻¹ NAA yielded the maximum number of shoots (16.35 ± 0.22 shoots per explant). Molecular analysis utilising 15 RAPD primers and 11 ISSR primers generated 84 and 76 scorable bands, respectively. The banding patterns across all 15 randomly selected regenerants showed no detectable polymorphism using RAPD and ISSR markers, indicating high genetic stability. The absence of somaclonal variation confirms that this optimised somatic embryogenesis protocol provides a reliable and scalable method for the mass propagation of true-to-type elite clones. This offers a highly effective biotechnological tool for both the sustainable commercial supply of pharmaceutical raw materials and the ex situ conservation of K. marginata germplasm.