Secoiridoid glycosides are naturally occurring phytochemicals of great importance for the food and pharmaceutical industry because of their various biological activities. Certain Gentiana and Centaurium species., which are recognized as the most important sources of these compounds, have become critically endangered due to overexploitation. In this study we describe a laboratory-scale approach for further implementation in large-scale production of secoiridoid glycosides, using a hairy root culture system of Centaurium maritimum L. Fritch, an underutilized and phytochemically unexplored species. Hairy roots were induced by Agrobacterium rhizogenes strain A40M70GUS and grown in Erlenmeyer flasks, as well as in RITA temporary immersion bioreactors (TIBs). About 2-4 times higher biomass production rate and up to 8 times higher total secoiridoid glycosides production rate were achieved in RITA bioreactors. Among the selected hairy root lines, line HR3 cultured in RITA (R) TIBs proved to be the most efficient considering both biomass and secoiridoid glycosides production rate. (C) 2013 Elsevier Ltd. All rights reserved.