We used the aseptic seedlings of 2 cultivars (Phalaenopsis ssp. cv. ‘Green Bear’ and Phalaenopsis ssp. cv. ‘Big Chilli’) to investigate the effect of LED in different light qualities on multiplication and rooting in vitro. The experimental design included 15 different light sources composed of six kinds of LED light, namely RED(630), R(630)8B2, R(630)7B3, R(630)5B5, R(630)3B7, R(630)2B8, BLUE, RED(660), R(660)7B3, R(660)5B5, CW5R(630)5, Warm W, R(630)6B3FR1, R(630)4B4FR2, R(630)3B6FR1, and WFL (control). Under red light illumination, seeding growth and the number of shoots were promoted, but chlorophyll content of leaves was reduced, but it was increased under the mixture of blue and red LEDs. Blue light had a positive effect on chlorophyll content. Root activity was increased with increasing ratio of blue in the combined red and blue LEDs and increased under the three light sources supplemented with far-red light. Blue and far-red light were more effective in root length. Shoots could be more successfully induced under Warm W light. Compared with WFL light, the number of shoots of the 2 varieties was increased by 53.17% and 46.37%, respectively. Therefore, the best light recipe for the multiplication and rooting of Phalaenopsis were Warm W and R(630)3B6FR1 light. The present study provides a proof of application of LEDs in Phalaenopsis tissue culture.