Plants are an important source of bioactive compounds, in particular Anti-cancer programs that may be linked to Plant products-phytochemicals Vinblastine and Vincristine mg/L are alkaloids derived from the Madagascar periwinkle plant Catharanthus roseus. These compounds have cell cycle-specific activity in the M phase, which is consistent with their ability to inhibit tubulin polymerization and prevent formation of the mitotic spindle. So in this study we evaluated the effects of several pathogenic and non-pathogenic fungi such as Alternaria alternate, Exserohilum rostratum on outcome production of Vinblastine and Vincristine in the Catharanthus roseus plant. The present study included several tests involved molecular diagnostic and assessment of fungal pathogenicity by using of polymerase chain reaction (PCR) and Bio-agent indicator techniques respectively, additional to estimation of outcome production of vinblastine and vincristine by using of High-performance liquid chromatography (HPLC) that is a chromatographic method used to estimate of vinblastine and vincristine after the plant infection of Catharanthus roseus plant with the fungi described above.Our result showed significant differences between the two fungi, as the results revealed that the fungus it was found that Alternaria alternate decresed the outcome of vinblastine and vincristine production (122.7mg/l and 153.8mg/l) respectively compared with control (150 and 160) mg/l respectively. Concerning Exserohilum rostratum the results were counterproductive, as the fungus in question increased the final product of vinblastine and vincristine. It was found that the amount was (175 mg/L and 181.3 mg/L) for vinblastine and vincristine respectively.