The Catharanthus roseus L. G. Don, commonly known as madagascar periwinkle, is a medicinally important plant which is of interest because it contains more than 130 terpenoid indole alkaloids (TIAs), some of which exhibit strong pharmacological activities. Two of these bioactive compounds, vinblastine and vincristine from C. roseus have been used extensively as anticancer drugs in the treatment of lymphoma and leukemia like. These compounds are produced in small quantities in the shoot tissues of periwinkle and expensive to harvest and purify. Several attempts has been performed to increase TIAs production by media optimization, precursor feeding and genetic manipulations of pathway genes. Despite significant efforts heretofore, limited success has been brought both in cell suspension and hairy root cultures of C. roseus . Increase in global demand for these two medicines have drawn attention to use biotechnological methods to increase the production of these important alkaloids in intact plant. TIA pathway is highly regulated, so that changes in precursor concentration or protein concentration are probably controlled by the plant phyto-hormones such as jasmonic acid (JA) and salicylic acid (SA). Therefore the aim of present study was to investigate the inducible effect of JA and SA hormones on expression of dat , d4h , str and g10h genes in TIAs pathway in shoot and root tissues and changes in accumulation of vinblastine and vincristine alkaloids which are products of this pathway. For do this, the total RNA was extracted from roots and leaves of plants at 0, 6, 24, and 72 hours after treatment with 50mM JA and 100mM SA hormones. cDNA was synthesized using specific primers and gene expression pattern was monitored by real time PCR technic. The result of thi study showed no production of vinblastine and vincristine in the roots which can be due to non expression of dat and d4h genes in this tissue. These genes seem to have an important role in production of these alkaloids. str and g10h genes were expressed in the root and shoot but the expression in the root is more than shoot. Analytical measurements indicated that both JA and SA hormones positively affect the expression of these four genes, though the effect of SA was significantly more than JA.Vinblastine and Vincristine alkaloids of treated plants were extracted by hollow fiber-based liquid-phase microextraction (HF-LPME) method and analyzed by high performance liquid chromatography (HPLC). Data analysis revealed an increase in vinblastine and vincristine alkaloids content after the treatment of hormones, which SA showed more pronounced effect. The most increase in both gene expression and alkaloids production was observed at 72 h after treatment of SA and JA. Based on the results of real-time PCR and HPLC experiments, it can be concluded that phyto-hormone treatment can increase expression of genes involved in TIAs pathway and consequently affect the production of vinblastine and vincristine anticancer alkaloids. Key words : Periwinkle, Vinblastin, Vincristin, jasmonic acid, salicylic acid, Real Time PCR, HPLC