Elicitors cause biosynthesis and accumulation of secondary metabolites by inducing defense responses. In this study, we treated the cell suspension cultures of Silybum marianum L. with MeJA (methyl jasmonate) (100 μM) and YE (yeast extract) (0.1 w/v) as elicitors and measured the content of Silymarin accumulation by HPLC (High Performance Liquid Chromatography). Accumulation of Silymarin significantly increased after 48 h of MeJA and YE application. In order to investigate the effect of abiotic (MeJA) and biotic (YE) stresses on expression of proteins in S. marianum cell suspension cultures, we employed high resolution two-dimensional gel electrophoresis coupled with MALDI-TOF-TOF (matrix assisted laser desorption ionization time of flightetime of flight) mass spectrometry. At least, 249 protein spots showed reproducible and significant changes in the gel. Spots were up or down regulated upon MeJA and YE treatments. Ten protein spots were identified using MALDI-TOF-TOF-MS. The identified proteins belong to different functional categories. The proteins were classified based on carbohydrate metabolism (spots 6 and 22), Nitrogen metabolism (spot 20), storage protein (spot 29), transport process (spot 12), protein modification and chaperones (spot 8), pathogenesis related (spot 1 and 2) and secondary metabolism (spot 3 and 53). The potential role of these proteins in the biosynthetic pathway of flavonoids and the finding of proteins in non-sequenced plants such as S.marianum requires further research.