The pollination biology of Notylia nemorosa was elucidated from field and ex situ observations. Field observations were carried out in Minas Gerais, southeastern Brazil, where this orchid is pollinated by males of Eulaema nigrita (Euglossini). Plants cultivated in Campinas (Sao Paulo State, southeastern Brazil) were eventually pollinated by males of Eulaema nigrita and Euglossa melanothricha (Euglossini). In both cases, Euglossini males collect aromatic compounds produced on the surface of the lip, fix the pollinaria on the dorsal surface of the labrum and perform pollination. This species is protandrous. Recently opened flowers present their stigmatic surfaces blocked and thus flowers act only as pollen donors. After 2-3 days the viscidium dries out and pollinaria cannot be removed anymore. Then, a narrow slit opens in the stigmatic cavity, allowing pollen deposition. Experiments performed with cultivated plants suggest that this orchid species is strongly, though not completely self-incompatible. Pollination experiments showed that bees tend to stay several minutes in each inflorescence. This behavior may promote some degree of geitonogamous pollination because flowers in male and female phases coexist in the same inflorescence. The combination of self-incompatibility with this specific pollinator behavior may explain the rarity of fruits in several neotropical reward-offering Orchidaceae.11611925