In Catharanthus roseus cell cultures, cytokinins (CK) improve monoterpenoid indole alkaloids (MIAs) accumulation. This metabolite production is correlated with an increase of transcripts corresponding to genes encoding both elements of the CK-signaling pathway and enzymes implicated in MIAs biosynthesis. However, it has not been demonstrated that the CK signal, leading to MIAs accumulation, comes through components identified as belonging to the CK-signaling pathway. In this work, we addressed this question, by transgenesis, using an inducible RNAi system targeting element of CK-signaling. In transgenic lines, the up-regulation by CK of two genes involved in MIA biosynthesis was abolished. These results demonstrate a relationship between the CK-signaling and the MIAs biosynthetic pathways.