This research aimed to measure the impact of provenance on the yield, chemical profile and antimicrobial activity of Rosmarinus officinalis essential oil (EO) collected from cultivated and wild plants from two different regions in Morocco (Fez and Figuig). The chemical composition analysis was accomplished utilizing the GC-MS techniques. Sixteen compounds were identified in the EO of both samples, dominated by 1,8-cineole (32.18%), camphor (16.20%) and α-pinene (15.40%) in cultivated type. The α-pinene (51.19) presents the majority compound in the rosemary samples collected from the wild populations.The antimicrobial activity was investigated by using the broth dilution methods against yeast, four bacterial strains and two molds. Rosemary aerial part provided EO with the highest yield and comes from Figuig. The GC-MS analysis demonstrated the existence of two chemotypes of oils: α-pinene as well as 1,8-cineole/camphor/α-pinene. Both EOs showed good antimicrobial activity against all microbial strains. The essential oil produced by Fez plants was the most effective against the selected microorganisms having MIC values of 0.315-2.5 mg/L.