<p>This study compared the effects of two culture systems, semi-solid (SS) and temporary immersion bioreactor (TIB®), on the micropropagation and morphoanatomical features of two banana varieties (‘Preciosa’ and ‘Thap Maeo’). The multiplication phase utilized Murashige and Skoog (MS) medium with 2.5&#xa0;mg L⁻¹ of 6-benzylaminopurine (BAP) in both systems. Following multiplication, elongation and rooting stages were conducted, and growth-related variables were evaluated after 40 d. Acclimatization was performed for 60 d, and morphoanatomical studies were carried out on banana leaves. Results showed that the TIB® system promoted greater height and biomass increment during multiplication, along with an increase in the number of leaves in ‘Preciosa’ variety. However, it resulted in lower multiplication rates. Specific anatomical modifications were observed in each variety. ‘Preciosa’ displayed enhanced functionality of adaxial stomata and increased abaxial stomatal density, while ‘Thap Maeo’ showed increased functionality of adaxial stomata and thicker palisade parenchyma. After acclimatization, the TIB® system led to greater height and diameter increments in ‘Preciosa’ plants, while the SS system improved the survival rate of ‘Thap Maeo.’ However, ‘Thap Maeo’ plants in the TIB® system exhibited lower survival rates due to issues such as etiolation and hyperhydricity. Regardless of the culture system, acclimatization for 30 d resulted in increased epicuticular wax deposition, higher stomatal density, and reduced stomatal functionality.</p>