The genus Hertia, which belongs to the Asteraceae family, is a flowering genus with 12 species found in Africa, North and South. Among the species present in Algeria, Hertia cheirifolia L. is distributed in the eastern regions of Algeria. The aim of this study is to evaluate its phytochemical composition with following pharmacological assessments: the antioxidant, antibacterial, and antifungal activities of Hertia cheirifolia L. essential oil (EO). GC-MS analysis was used to analyze the chemical constituents of H. cheirifolia essential oil. The antioxidant capacity was assessed using DPPH, FRAP, and H2O2 tests. The EO was also tested for its ability to inhibit six strains of microorganisms, including two Gram (+) and four Gram (-) strains. The antifungal activity was tested by analyzing the effect of the EO on the mycelial growth of Fusarium oxysporum f.sp. lycopersici (FOL) fungi. Results showed that primary volatile components were α-pinene (32.59%), 2-(1-cyclopent-1-enyl-1-methylethyl) cyclopentanone (14.62%), (-)-germacrene D (11.37%), and bakkenolide A (9.57%). H. cheirifolia EO showed inhibitory effects against DPPH, H2O2, and FRAP (IC50 = 0.34 ± 0.1, 0.053 ± 0.1, and 0.047 ± 0.01 mg mL-1, respectively). The EO also exhibited moderate antibacterial effects against Staphylococcus aureus ATCC 25923 (S. aureus), Streptococcus pneumoniae ATCC 49619 (S. pneumoniae), and Enterobacter aerogenes ATCC 13048 (E. aerogenes), as well as significant antioxidant potential and varied antifungal activity based on dosage and fungal strain. To our knowledge, no previous research has examined the antifungal capacity of H. cheirifolia oil and oil-mycelial development of the FOL relationship. To fully explore the benefits of H. cheirifolia EO, more in vivo research is necessary, along with more testing on other bacterial and fungal strains.