<p>Light-emitting diodes (LEDs) are increasingly used in horticultural studies to control light quality. <i>Spathiphyllum cannifolium</i>, a species widely cultivated as an ornamental plant, uses tissue culture as one of its primary propagation methods. However, similar to other plants, in vitro rooting presents common challenges for the multiple propagated shoots of <i>S. cannifolium</i>. Therefore, in this study, an ex vitro rooting method was applied in a microponic system to elucidate the effects of LEDs on rooting efficiency by investigating plantlet growth, photosynthesis, and sucrose metabolism. The results showed that, compared to red (R) and blue (B) LEDs, as well as fluorescent light, the combination of R and B significantly (<i>p</i> &lt; 0.05) enhanced plantlet growth, as evidenced by higher values in leaf number, leaf area, shoot height, shoot dry weight, root number, and root dry weight. Photosynthetic parameters, including net photosynthetic rate, stomatal conductance, and transpiration rate peaked under R + B, while intercellular carbon dioxide concentration decreased. Additionally, the R + B treatment resulted in the highest chlorophyll content and sucrose accumulation, along with increased activities of key enzymes involved in sucrose metabolism, such as sucrose phosphate synthase (EC 2.4.1.14) and sucrose synthase (EC 2.4.1.13), and decreased activities of acid invertase (EC 3.2.1.26) and neutral invertase (EC 3.2.1.26). These findings suggest that the combination of red and blue LEDs is the most effective in promoting <i>S. cannifolium</i> growth and physiological functions during ex vitro rooting and indicate the potential for optimizing plant development in controlled environments.</p>