The aim of the present work was to evaluate the effect of 2,4-D and BAP on the protocorm formation from meristematic areas of the root and leaf segments of Miltonia flavescens Lindl., as well as to test the effect of sucrose concentrations and pH levels on the in vitro growth of protocorms. Root apexes and leaf segments of in vitro-germinated plants were cultured on MS/2 culture medium supplemented with 2,4-D and BAP and kept in the dark for 210 days. Explants were then transferred to light and cultured on culture medium lacking growth regulators. Percentage of explants with protocorms and average number of protocorms regenerated per explant were evaluated. The combination of 3 mg L-1 2,4-D and 1 or 3 mg L-1 BAP provided the best results for protocorm formation. Protocorms were cultured on MS/2 culture medium to test the effect of sucrose (0, 15, 30, 45, and 60 g L-1) and pH (5.0, 5.8, and 6.6) on their growth. The concentrations of 15 and 30 g L-1 sucrose, independently of the pH, were more efficient for the in vitro culture of this species, promoting an increase in the number and length of the roots, plant height, and fresh weight. The pH was significant only in combination with 15 g L-1 sucrose for the variables length of the largest root and fresh weight.
Orchidaceae; growth regulators; sucrose; pH; Miltonia flavescens
