Light-emitting diodes (LEDs) have become very popular for the production of horticultural crops. LEDs represent an alternative lighting source to regular fluorescent (FL) bulbs, increasing the quality of plants and minimizing production costs. LEDs also provide selective light intensity and quality, suitable for commercial micropropagation. The objective of this study was to evaluate the growth and development of in vitro ornamental bananas under different light sources. Two ornamental banana varieties were selected for this study: Musa ‘Little Prince’ and Musa ‘Truly Tiny’. Light quality and intensity of three different light sources were evaluated: LED-1 (116 μmol m−2 s−1), LED-2 (90 μmol m−2 s−1), and FL (100 μmol m−2 s−1). Length and biomass of plantlets were greater under LED-1 compared to FL but not significantly different from LED-2. The fresh and dry weight of shoots and roots, number of leaves, and number and length of roots were not significantly different between treatments. Chlorophyll content was greater under LEDs. Leaf number and stomata number and size were greater under FL. Our results indicate that shoot length and biomass could be improved by optimizing light quality and intensity. Different responses to light sources between the two banana varieties also indicated a genotype effect.