This study was investigated to develop efficient protocols for adventitious and hairy root cultures of Plumbago zeylanica. Adventitious roots were initiated from leaf and stem explants cultured on MS medium with different concentrations and combinations of auxins. The highest number of roots was obtained when the explants was cultured on MS medium with 1.0 mg·L-1 IBA and 0.5 mg·L-1 NAA. Hairy root culture of P. zeylanica was obtained by infecting leaf explants cultured in vitro with Agrobacteriumrhizogenes MTCC 532. The highest frequency of explant transformation was about 93%. The developed culture exhibited fast growth and high lateral branching on growth regulator free MS medium. The rootcultures obtained were inoculated into B5, MS and SH media supplemented with different carbon sources with or without auxins and were placed on a rotary shaker 80 rpm for 35 days under dark or light conditions. Of the three media tested, MS medium sustained better root growth than others and sucrose proved to be the best carbon source. The biomass in hairy root culture was higher than in nontransformed root culture