Centaurium erythraea plants obtained by indirect organogenesis are described in the paper. The plants were initiated from a single adventitious shoot regenerated from callus derived from the cotyledon of a 30-day-old seedling. The shoot was multiplied on MS medium supplemented with IAA (0.1 mg·L-1) and BAP (1.0 mg·L-1). The multiplication rate (28 shoots per culture within 4 weeks) was highest at the first subculture and decreased in further subcultures. The shoots were rooted on MS medium. The effect of IBA (0.1 mg·L-1) on the number of shoots forming roots differed depending on the composition of the basal medium (MS). The rooted shoots were transplanted to soil and grown in a greenhouse with 90% effectiveness. RAPD analysis was done with adventitious shoots of C. erythraea from in vitro culture. In shoots and whole plants regenerated from the callus tissue, secoiridoid content was determined by the HPLC method. We showed significant differences in morphology (leaf size, fresh and dry weight and height of plants) and changes in the DNA profiles as compared to earlier reports for shoot tip-derived shoots and plants of C. erythraea, but the two groups of plants biosynthesized the same qualitative pattern and similar levels of secoiridoids, up to 150 mg·g-1 dry weight; the increased biomass of plants regenerated from callus tissue makes them a better source of secondary metabolites.