The aerial parts of Artemisia annua L. were collected from Varzob, Rudaki, and 1-lisor regions of Tajikistan. The essential oil was obtained by hydrodistillation and analyzed by gas chromatography-mass spectrometry. The essential oils of aerial parts of A. annua were dominated by the monoterpenoids like camphor (32.5%-58.9%), 1,8-cineole (13.7%-17.8%), camphene (4.5%-8.4%), and alpha-pinene (1.9%-7.3%). Hierarchical cluster analysis of A. annua essential oils indicated the existence of 3 A. annua chemotypes: camphor/1,8-cineole, camphor, and artemisia ketone. The essential oils of A. annua show weak antioxidant activity and average antibacterial activity. In our opinion, the antibacterial activity of A. annua essential oils is related to the presence of 1,8-cineole. To our best knowledge, this is the first report concerning the chemical composition, chemotypic variation, antioxidant, and antimicrobial activities of the essential oils obtained from the aerial parts of A. annua, growing wild in Tajikistan.