Rhynchostylis retusa, popularly known as the Foxtail Orchid, is an epiphytic orchid of significant ornamental and medicinal value. Due to
indiscriminate collection and habitat destruction, its natural populations are declining, necessitating efficient in vitro micropropagation
protocol for conservation and commercial production. The present study established a rapid and reproducible micropropagation protocol
using shoot meristem and leaf explants excised from in vitro raised plantlets. Murashige and Skoog (MS) medium supplemented with
various concentrations of Plant Growth Regulators (PGRs) was used. shoot meristem segments exhibited direct organogenesis, yielding
the highest number of multiple shoot buds (6.3±0.67 shoots/explant) on MS medium fortified with BAP (2.0 mgL-1) and NAA (1.0 mgL-1).
Conversely, leaf segments underwent somatic embryogenesis, producing the maximum percentage (77.33±1.96%) of Protocorm-like
bodies (PLBs) on the same hormonal regime. For shoot elongation, agar solidified MS medium supplemented with BAP (1.5 mgL-1) and NAA
(0.9 mgL-1) proved better than liquid nutrient media, achieving a shoot length of 2.84 ±0.04 cm. Rooting was best induced on half strength
MS medium supplemented with a combination of IBA (0.5 mgL-1) and NAA (0.5 mgL-1), resulting in an average of 5.17 roots per plantlet. The
plantlets were successfully acclimatized with a survival rate of 70% in a potting mixture of coconut coir, sawdust, and coal. This protocol
offers a viable pathway for the mass propagation and ex situ conservation of R. retusa.