The results showed the ability of the Axillary Bud explants to produce higher number of shoot lets without intervening callus phase, where all the plantlets were uniform in height and growth. This study establishes a simple, rapid, high frequency micropropagation method for Dendrobium barbatulum Lindl. from axillary bud explants and also this present system of normal root culture would be beneficial for the sustainable utilization of this rare endemic medicinal plant for its bioactive ingredients, thereby providing an alternative method rather than destroying whole plants that are not under cultivation. From these studies it can be concluded that B5 medium suitable for Dendrobium barbatulum Lindl.. This study also revealed that a low concentration of 0.5 mg BAP + 3 mg NAA + 50 ml CM was found to be more suitable for plantlets and multiple plantlets. B5 medium supplemented with Basal B5 Medium + 1 mg BAP+ 3mg IAA+ 50 ml CM +500 AC giving highest percentage was found to be suitable for In vitro Rooting