OPTIMIZATION OF TISSUE CULTURE PROTOCOL FOR REGENERATION AND DIRECT MULTIPLICATION OF MORINGA OLEIFERA BY SEED GERMINATION

Moringaoleifera(G.H.No.:95436,Dr.MuneebaKhan,CentreforPlantConservation,University of Karachi) known as the miracle tree, is a plant of great importance. The plant is well known for its diverse medicinal and nutritional properties. The objective of the research is focused on tissue culture of Moringaoleifera through callus induction, direct multiplication as well as from seed culture.Thestudyisalsofocusedonoptimizationofsterilizationprotocolandmediaforthetissue culture processes. As expected, Moringaoleifera gave promising results using MS Media supplementedwithdifferentconcentrationsofPlantGrowthRegulatorssuchasIAA,NAA,BAP, Kinetin and 2,4-D. Seed initiation gave optimum results on MS Basal media. Direct multiplication of the plant was observed with optimum results on MS media supplemented with 0.1 mgL-1BAP. For callus induction, effects of Plant Growth Regulator and photoperiod were observed. In Dark conditions, maximum growth was seen on MS media supplemented with 2, 4-D (2 mgL-1), and that in light conditions, MS media with 2,4-D (2 mgL-1and 3 mgL-1) gave almost similar results. Initially, seed coats were removed inside the Laminar Flow Hood but this was affecting the sterilization negatively. Seed sterilization was then modified by removing seed coats and sterilizing them outside the Laminar Flow Hood. The seeds were then transferred to the LFH for furthersterilization,andthenculturedonthe growthmedia.Forthesterilizationoftheexplantfor directmultiplicationandcallusinduction,firstlyEthanolwasusedwhichcausedbleachingeffect. Later, Ethanol was removed from the protocol, reducing the bleaching effect to zero, and sterilization was maximized.