Ex-Situconser Ex-Situconservation of Protectedmedicinal Plants:Valeriana Jatamansi Jones Andrauvolfia Serpentinal. Benth.Ex Kurz. By Tissue Culture Technique2

Abstract

Subculture of in-vitro nodes ofValeriana jatamansii Jones was carried out for sixconsecutive batches or passages on MS medium supplemented with 1 ppm BAP and0.5 ppm NAA. Optimum number of shoots (average 11 shoots) was obtained in thefifth batch. The shoots were acclimatized and hardened in three different media:cocopit, sand and the mixture of sand and soil (1:1 v/v). The highest percentage ofsurvival in-vivo(91.6%) was achieved in cocopit followed by sand (83.3%). Theshoots and their roots appeared healthy and longer in cocopit. However, the in-vitroplants hardened in sand adapted to the soil relatively faster and with over 95%survival rate. The density of roots was relatively higher in the plants hardened in sand.The size of the rhizome seemed bigger in those plants which were hardened in sandand then transferred them into the soil. The in-vitroraised plants hardened in cocopitwere slow and ineffective for rapid adaptation in the soil. Nodes from the in-vitro developed shoots of Valeriana jatamansii Jones weresubcultured in MS medium supplemented with four different concentrations of NAA:0.5 ppm, 1 ppm, 1.5 ppm and 2 ppm NAA for the production of root mass. profuse ofroot mass of root was observed in MS + 0.5 ppm NAA. Next, the most effectivestrength of MS for the optimum proliferation of hairy roots was verified byinoculating the in-vitrodeveloped nodal explants in four different strengths of MS medium: 41 MS, 21 MS, 43 MS and full strength of MS, supplemented each with 0.5 ppm NAA. Maximum proliferation of hairy roots was observed in NAA. 21 Next, shoot tips and nodes from thein-vivoplants ofRauvolfia serpentinaL. Benth.ex Kurz were inoculated on MS media with fifteen different combinations andconcentrations of BAP and NAA. Shoot tip explants responded with healthy anddistinct multiple shoots in MS + 2 ppm BAP + 0.5 ppm NAA. Multiple shoots werediffirentiated from the callus. The nodal explants differentiated into, healthy andrelatively sturdy shoots in MS + 0.5 ppm BAP + 0.5 ppm NAA. There was no callusformation. This protocol may be appropriate to produce genetically homogenous progeny of the plant. Shoot tip explants differentiated optimum callus, without anyshoot and leaf, in MS + 0.5 ppm BAP + 1 ppm NAA. Nodal explants produced soft,dark-brown callus with upper green surface in MS + 1.5 ppm BAP + 1 ppm NAA.Single shoot appeared fromthe callus.