Axillary buds of Japanese persimmon (Diospyros kakiThun. cv. ‘Hiratanenashi’) were decontaminated, their budscales removed aseptically and placed in modified Murashige and Skoog (MS) medium containing 4.5 μMzeatin. Over 60% of the buds initiated shoots in 6 weeks. A multiplication rate of over 10-fold could be achieved in 6–8 weeks by two transfers (shoot proliferation and shoot elongation) in the same medium. Attempts to initiate shoots from meristem tips were unsuccessful. Using shoots that are > 25 mm long, c. 85% rooting was accomplished over a 6-week period in diluted (nitrogen content, 1/4 strength; other nutrients, 1/2 strength) MS medium containing 2% activated charcoal and 4.9 μMindole-3-butyric acid (IBA) with a liquid overlay of the undiluted medium containing 4.5 μMtrans-zeatin (t-Z). Techniques are also described for the regeneration of whole plants from leaf segments (from in vitro propagated plants) using a combination of t-Z (10 μMand anyone of three auxins tested—indole-3-acetic acid, IBA or (α-naphthalene acetic acid (0.001-0.01 μM). The presence of auxins was not essential but slightly accelerated the organogenic callus formation and organogenesis. Rooted plantlets were planted into a mixture of peat, perlite, and CaCO3and acclimatised in a fog tent for 4–8 weeks.