Species from the genus Gentiana (fam. Gentianaceae) are a valuable source of secondary metabolites: secoiridoides glucosides, xantones and other compounds that have commercial application. Gentiana pneumonanthe is a perennial plant, 30 - 70 cm tall. In the wild it grows in boggy meadows, fens, moist heaths and sandy or peaty clearings in woods, sometimes on acid soils. Its flowering season is from July to August. The species is critically endangered in Serbia. In vitro cultures of many gentians are already established in our laboratory. Here we present results of in vitro flowering shoot cultures of G. pneumonanthe. Vegetative shoot cultures were obtained from seedlings and buds from adult plants. They were grown under long day illumination, on 1/2 WPM media containing cytokinin and auxin. After several subcultures, shoots formed flower buds and subsequently flowered in vitro. Flowering shoot segments showed a vertical gradient in morphogenic capacity for callus, root and/or flower bud formation. This material seems to be a suitable system for fertilisation in vitro and seeds production as well as genetic manipulations.