Blackstonia perfoliata (L.) Huds. (fam.Gentianaceae) is annual plant, 10-40 cm in height. This critically endangered plant grows at wet, sandy and slightly salty meadows. The habitats of this weak competitor are also clayly and calcite rich soils, and wet alluviums. As a representative of the Mediterranean areal type, the plant reaches the altitude of 1300 m above sea level, but it is more spread in the plains. Yellow wort contains gentiopicroside, so it is considered as a substitute for Centaurii Herba.

In vitro root culture of B.perfoliata was initiated on MS medium supplemented with different concentrations of auxins and cytokinins. The experiment was performed to investigate the effects of various growth regulators in in vitro plant regeneration. In the presence of cytokinins numerous adventitious buds were formed, which further developed into shoots. Auxins significantly decreased number of buds, but in their presence numerous root branches were developed. Various root parts, used in this experiment, were equally effective in adventitious buds formation. On hormone-free medium shoots were successfully rooted, plants were grown and flowered. Those plants gave viable seeds, which produced F1 generation. In vitro root culture is convenient method for plant regeneration and may be the suitable system for transgenic plant production.