Vanda tricolor is endemic orchid which lives on the slopes of Mount Merapi. The use of medium and plant growth regulator in multiplication of in vitro cultures will increase the growth of Vanda tricolor orchids. The purpose of this study was to determine the best concentration of 2,4-D on New Dogashima Medium and Foliar Fertilizer for Orchid Vanda tricolor embryosomatic induction. The study was arranged in a single completely randomized design (CRD) consisting of 6 treatments, namely without 2.4 D, 2.5 mg / l 2.4 D, 5 mg / l 2.4 D in the New Dogashima mMedium and Foliar Fertilizer. Each treatment consisted of 3 replications, each replication consisted of 3 samples, so that the total number was 54 units, each treatment was supplemented with BAP 0.5 mg / l. The results showed that Foliar Fertilizer was able to substitute New Dogashima Medium and a concentration of 2.5 mg / l 2.4 D gave the best results for the percentage of callus explants (22.22%), the callus emergence (9.7), and the embryo emergence (6,7).