In Catharanthus roseus, the first step of monoterpenoid indole alkaloids (MIA) biosynthesis results from the condensation of the indole precursor tryptamine with the terpenoid precursor secologanin. Secologanin biosynthesis requires two successive biosynthetic pathways, the plastidial methyl-D: -erythritol 4-phosphate (MEP) pathway and the monoterpene secoiridoid pathway. In C. roseus cell culture, the expression of several genes encoding enzymes of these two pathways is dramatically down-regulated by auxin, while strongly enhanced by cytokinin and methyl-jasmonate. Furthermore, our previous studies have shown that protein prenylation events are also involved in the transcriptional activation of some of these genes. In the present work, we investigate the involvement of protein prenylation in the jasmonate signalling pathway leading to MIA biosynthesis. Inhibition of protein prenyltransferase down-regulates the methyl-jasmonate-induced expression of MEP and monoterpene secoiridoid pathway genes and thus abolishes MIA biosynthesis. Jointly, it also inhibits the methyl-jasmonate-induced expression of the AP2/ERF transcription factor ORCA3 that acts as a central regulator of MIA biosynthesis. Finally, a specific silencing of protein prenyltransferases mediated by RNA interference in C. roseus cells shows that inhibition of type I protein geranylgeranyltransferase (PGGT-I) down-regulates the methyl-jasmonate-induced expression of ORCA3, suggesting that PGGT-I prenylated proteins are part of the early steps of jasmonate signalling leading to MIA biosynthesis.