Mass propagation of Satyrium nepalense D.Don.—A medicinal orchid via seed culture

An in vitro plant regeneration protocol was successfully established in Satyrium nepalense, a terrestrial orchid by culturing immature seeds from unripe fruits. Seeds were germinated on Murashige and Skoog, Knudson C and Knudson C modified Morel medium. The germination of the seeds and development of protocorm was highest in MS medium (86.7%) followed by Knudson C modified Morel medium (74%) and Knudson C medium (61.2%). Among the cytokinins tried for multiple shoot induction from the protocorm, 1-phenyl-3-(1,2,3-thiadiazol-5-yl)-urea (TDZ) was found to be superior. Indole-3-butyric acid (IBA) was effective for inducing healthy roots. Well-developed plantlets were hardened in vermicompost (leaf litter + cow dung 1:1), sand and coconut husk (1:1:1).