Cattleya trianae is an endemic species from the tropical rainforest in the Colombian Andes. Its survival is currently threatened due to habitat loss and commercial overexploitation. This study evaluates ten substrates, some organic (pine bark, coconut fiber and wood shavings), some inert icopor (polystyrene foam), vegetable coal and their combinations, and the effects these have on morphometric and phenotypic traits in the hardening phase of 250 plants of C. trianae cultivated in vitro. Recorded data include percent survival, length of longest leaf, biomass (wet weight) and number of roots and leaves at the beginning and at the end of the experiment. After the hardening phase, the plants were taken to a greenhouse and later to the natural environment. Coconut fiber alone or mixed in equal parts with pine bark and coal was the most efficient substrate when percent survival (80±SE=0.3742), biomass, and leaf length were evaluated. Hardened plants displayed qualitative characteristics such as vigor, hardness and waxy texture, strength of green coloration in the leaves, and velamen formation. Under greenhouse conditions, plants grew better with filtered light, relative humidity bordering on 80 %, permanent aeration, misting with water, and an average temperature of 25±2 °C. Invigorated plants were firmly anchored on their host trees.