Background: Acampe ochracea (Lindl.) Hochr. is an epiphytic orchid of significant ornamental and medicinal value distributed across Bangladesh and Southeast Asia? Due to habitat destruction, over-collection and slow natural propagation rates, its population is declining. Conventional propagation methods are insufficient to meet commercial and conservation demands.

Methods: This study established an efficient in vitro micropropagation protocol using nodal and leaf segments excised from in vitro raised seedlings. Explants were cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of 6-Benzylaminopurine (BAP), Kinetin (Kn), α-Naphthaleneacetic acid (NAA), Indole-3-acetic acid (IAA) and Picloram (Pic) to induce direct organogenesis and protocorm-like bodies (PLBs). Subsequent shoot elongation was evaluated in both solid and liquid MS media, followed by rooting in auxin enriched media and acclimatization.

Results: The highest frequency of multiple shoot buds (MSBs) from nodal segments (93.33 ± 1.72%) with a maximum number of shoots (6.2 ± 0.58 per explant) was achieved on MS medium supplemented with 2.0 mg/l BAP + 1.0 mg/l NAA. Leaf segments produced the highest percentage of green PLBs (77.33 ± 1.96%) on the same medium. For shoot elongation, solid MS medium fortified with 1.5 mg/l BAP + 0.9 mg/l NAA proved superior, yielding shoots of 2.81 ± 0.03 cm. Rooting was optimal on MS medium supplemented with 0.5 mg/l IAA + 0.5 mg/l NAA, producing 5.17 ± 0.40 roots per shoot. The survival rate of transplanted plantlets was 70% during acclimatization.