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Multiple shoots of Chinese chestnut (Castanea mollissima Blume), produced in vitro from axillary buds of juvenile shoots, were successfully rooted and transplanted to the greenhouse. Benzyladenine (BA) at 0.44 μM promoted the proliferation of axillary shoots, but 4.44 μM and 44.4 μM of BA inhibited buds from sprouting and promoted callus growth. Zeatin at 4.56 μM, induced longer and more vigorous shoots than BA, but did not promote the multiplication of axillary shoots. Rooting was achieved by a 1 sec basal dip of excised microshoots in 9.8 mM or 14.8 mM indolebutyric acid (IBA) solution. The treated shoots were transferred to a plant growth regulator-free medium in flasks or to plastic flats containing sand under high humidity. Roots developed within 30 days in both rooting regimes.