Abstract An efficient seed germination system was developed for an endemic and endangered orchid of the prairies of Bogotá, Colombia. The effects of three culture media [Murashige and Skoog (MS) salts (1962); Knudson C (KC) salts (1946); and Hydro-Coljap® salts], activated charcoal [0 and 0.5% (w/v)], α-naphthaleneacetic acid (NAA; 0.0, 2.68 and 5.37 μM), and four light regimes (white light, darkness, red light, and far-red light) on asymbiotic germination of Odontoglossum gloriosum, were studied. The best germination percentage and germination time was obtained on agar-solidified medium supplemented with Hydro-Coljap® salts and 2.68 μM NAA under red light with a 16-h photoperiod. The addition of activated charcoal did not stimulate either germination or the development of O. gloriosum seedlings. This study describes a single medium-based protocol able to achieve more than 330 000 seedlings within 40 wk, starting from a single capsule. This protocol is sufficient for both large-scale propagation and in vitro conservation of this threatened orchid.