Abstract: Many Orchidaceae, especially those occupying periodically dry, epiphytic microhabitats in the humid tropics, are believed to engage in the water-conserving crassulacean acid metabolism (CAM) photosynthetic pathway. However, the photosynthetic pathway has been studied in only c. 5% of all orchid species. Here we extend the survey to 1079 orchid species, mainly from Colombia, by assessing the presence of CAM based on the carbon isotopic signature (δ 13C values) of herbarium specimens. Ninety-six species, representing 8.9% of those analysed, had δ 13C values less negative than −20‰, indicating CAM. Epiphytism was the predominant life form (75.2% of species sampled), and 9.4% of these epiphytes showed a CAM-type isotopic signature. Isotope values suggested CAM in 19 terrestrial orchid species, 14 species from high elevation (2000–3400 m) and species from six genera that were previously unknown to engage in CAM (Jacquiniella, Meiracyllium, Pabstiella, Psychopsis, Pterostemma and Solenidium). We conclude that CAM is the major pathway of carbon acquisition in a small but broadly distributed fraction of tropical orchids and is more prevalent at lower elevations.