The research aimed to explore the active ingredients of rue extract and essential oil as an ornamental and medicinal plant species and to assess their antimicrobial activity. So, rue shoots were collected from the natural habitat of this species in the elevations of Chaboksar, Guilan province in October and November 2019. The leaves and stems were oven-dried at 75 °C for 24 hours, and their extracts and essential oils were extracted with a Clevenger. The active ingredients of the extracts and essential oils were detected with GC-MS. The most abundant compounds in the leaf essential oil, stem essential oil, leaf extract, and stem extract were found to be hexadecanoic acid, isomaturnin, 2-ethyl-1,3,4,5,6,7,8-heptamethy, and 12-methoxy-19-norpodocarpa, respectively. The microbial assay by the disk diffusion method on the strains of Pseudomonas aeruginosa, Staphylococcus aureus, Acinetobacter baumannii, and Escherichia coli showed that the leaf and stem extracts were most influential in inhibiting A. baumannii. The assessment of MIC and MBC revealed that S. aureus was the most sensitive bacterium to the leaf and stem essential oils with the lowest MIC and MBC and A. baumannii was the most sensitive bacterium to the leaf extract with the lowest MBC. So, rue extract and essential oil can be used as an antimicrobial compound against human pathogenic bacteria. Manuscript profile