An efficient protocol for in vitro propagation of gloxinia has been established in this study. Adventitious shoot regeneration was compared among various leaf-derived explants cultured on MS medium supplemented with different concentrations of BA, TDZ and NAA for investigating optimal explants and size of leaves for obtaining efficient shoot regeneration. Different organogenesis programs including shoot regeneration, caulogenesis and rhizogenesis were firstly observed within 3 weeks of culture. A great number of vigorous shoots were directly obtained from in vitro intact leaves, leaf laminas and leaf segments. Results showed that leaf laminas when cut into 6 parts were optimal for in vitro multiplication with a great number of newly forming shoots (46.2 shoots/explant) obtained in medium supplemented with 1.0 mg l^-1 BA and 0.2 mg l^-1 NAA, giving rise to a 100% of shoot formation within 4 weeks. Shoots were vigorously rooted on medium containing 0.2 mg l^-1 BA and 0.1 mg l^-1 NAA after 4 weeks of culture, and plantlets were then successfully acclimatized in greenhouse. Results from this study reveal a potential procedure for plant production simply via leaf culture and promise an efficient approach for transformation technique of this valuable plant because of its capacity of direct regeneration from leaf explants.