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The main aim of present study was to overcome the problems associated with the in vitro culture initiation in Terminalia arjuna. The micropropagation of tree species is not easy as shrubs and herbs. Many problems encountered from explant collection to in vitro culture establishment. The problems that have been occurred during T. arjuna micropropagation were culture contamination, phenolic exudation, bud growth inhibition, shoots yellowing and leaf fall. All these problems have been solved by applying certain treatments prior to explant collection and inoculation. The mother tree was lopped in November months (six months prior to explant collection) to remove any inhibitory substance and release bud growth. Different sterilizing agents were used to minimize the bacterial and fungal contamination. Some modification in culture media (use of different concentration of NH4NO3 and KNO3 salts and adenine sulphate) was done. Surface sterilization of nodal explants collected from lopped branches with 0.1% HgCl2 for 8 min., treatment with chilled antioxidant solution (Ascorbic acid, Citric acid and PVP) and half strength of NH4NO3 and KNO3 salts of MS medium supported 100% bud break response with proliferation of green and healthy in vitro shoots. Removing these hurdles already in the initial stage of micropropagation is very important and maximize mass in vitro propagation of this medicinally important Arjun tree. Original Research Article Choudhary et al.; ARRB, 35(12): 154-160, 2020; Article no.ARRB.64676 155

Problems Encountered during In vitro Culture Establishment in Terminalia arjuna