Catharanthus roseus is a medicinal plant containing very important alkaloids such as Vincristine and Vinblastin that are used to treat a variety of cancers. Due to the importance of the plasticity genes such as rolC in the biosynthesis of alkaloids, this study we investigated possible improvement of rooting and root induction in C. roseus using the rolC gene. To this end, the seeds of this plant were superficially sterilized and cultured in in vitro conditions. The leaf explants of these in vitro plants were inoculated by Agrobacterium tumefaciens carrying pBI121-rolC plasmid (rolC under control of the CaMV 35S promoter). The inoculated explants were cultured in five different media with or without hormones for rooting. After nine days, the first roots appeared in leaf explants on the MS medium without hormones and containing cefotaxime antibiotic. Two lines had very rapid rooting and the roots branched quickly. Molecular analysis by PCR using rolC specific primers confirmed the presence of the rolC gene in the transgenic plants. Results showed that the rolC gene could be used to induce rooting in the medicinal plants and that the rolC gene is also more successful in induction of hairy roots than various different combinations of plant hormones.