The decline of wild orchid populations derives from the clandestine looting and fragmentation of their natural habitat. In vitro orchid seedling propagation can represent a solution to this problem. This study aimed to establish an in vitro propagation system in Myrmecophila grandiflora, from germination in a semi-solid medium to development in a liquid medium, using temporary immersion (TI) techniques. For in vitro germination of the Myrmecophila grandiflora seeds, effect of different combinations and concentrations of full- and half-strength Murashige and Skoog medium (MS) supplemented with 6-benzylaminopurine (BAP, 0, 0.5, and 1 mg l^-1) were researched. Different immersion frequencies (every 4, 8, and 12 h) and volumes of liquid culture medium (5, 10, and 20 ml of medium per explant) were evaluated for in vitro development, using the ebb-and-flow bioreactor. In the semi-solid medium, for researching of germination percentage of seeds at seven weeks of culture, and in the liquid medium we used 0.5 cm seedlings in TI to evaluate their development. All treatments had germination percentages greater than 89%. The best development in TI occurred with an immersion frequency of 2 min every 4 h, with 10 ml medium per explant.