Rosa damascena Mill. is a very fragrant member of Rosaceae family, exhibiting strong antioxidant and antimicrobial activity. In this study, in vitro cell cultures from Rosa damascena Mill. flower petals were established and the effects of precursor L-phenylalanine, the elicitor methyl jasmonate and light/dark treatments on the accumulation of characteristic rose volatile compounds, tocopherols and beta-carotene in addition to the antibacterial activity of petal callus extracts were investigated with Solid-Phase Microextraction Combined with Gas Chromatography-Mass Spectrometry (GC-MS). The amount of beta-pinene increased by 236 times when 500 mu M Phe was added to the culture medium or under light treatment. Strong antibacterial activity from petal callus extracts, up to 2.6-fold compared to amikacin as well as original petal tissues, was obtained under dark treatment for P. aeruginosa ATCC 27853 and S. aureus ATCC 25923. These results revealed that petal callus extracts are sources of natural antioxidants and antibacterial compounds.