<p>The present study focused on the micropropagation of three cultivars of <i>Begonia rex</i>, namely ‘Silver Dollar’, ‘Jurassic’, and ‘Silver King’. The research aimed to assess the effectiveness of various plant growth regulators (PGRs) in inducing callus formation and direct bud induction in leaf explants, as well as to determine the most suitable potting substrates including a mixture of cocopeat and perlite (CP + P) (1:1, w/w), peat moss and perlite (PM + P) (1:1, w/w), leaf mold (LM), loamy sand soil (S), and a combination of loamy sand soil and leaf mold (S + LM) (1:1, w/w) for acclimatization of the plantlets of three cultivars. The leaf explants were cultured on Murashige and Skoog (MS) medium, supplemented with different concentrations of 1-Naphthaleneacetic acid (NAA), Indole-3-butyric acid (IBA), thidiazuron (TDZ), and 6-benzyladenine (BA). Results showed that the medium containing a combination of 0.05 mg L⁻¹ BA and 0.2 mg L⁻¹ TDZ exhibited the highest percentage (22.00%) of direct bud formation per leaf explant. However, all treatments showed some level of indirect bud formation, with percentages ranging from 33.0% to 100%. The most significant bud proliferation (100%) was observed in the medium with 1.0 mg L⁻¹ IBA and 1.0 mg L⁻¹ BA, specifically in the ‘Jurassic’ cultivar. Interestingly, the acclimatization process of the plantlets yielded varied results for each cultivar when cultivated in each substrate. In terms of having higher values of the Selection Index of Ideal Treatment (<i>SIIT</i>) parameter calculated from 10 different features, both CP + P and LM substrates showing promising results for ‘Silver Dollar’ and CP + P substrate for ‘Jurassic’, while S and CP + P performed well for the ‘Silver King’ cultivar. These findings provided valuable insights for gardeners and researchers looking to efficiently propagate and acclimatize <i>B. rex</i> cultivars.</p>