| Summary: | The application of natural products for pest control is important in modern farming. In the present study, <i>Artemisia santonicum</i> L. and <i>Artemisia lerchiana</i> Weber essential oil and exudate profiles were determined, and their potential as inhibitors of seed germination, acetylcholinesterase, and phytopathogenic mycelium growth were evaluated. Essential oils (EO) were obtained via hydrodistillation and exudates (AE) by washing aerial parts of the species with acetone. EO and AE’s composition was identified using GC/MS. Eucalyptol (1,8-cineole) and camphor were found to be the main components of <i>A. lerchiana</i> EO, while β-pinene, <i>trans</i>-pinocarveol, α-pinene, α-terpineol, and spathulenol were established as major compounds of <i>A. santonicum</i> EO. Strong inhibition on <i>Lolium perenne</i> seed germination was found at 2 µL/mL and 5 mg/mL using aqueous solutions of EO and AE, respectively. An inhibitory effect on acetylcholinesterase was established, with an IC<sub>50</sub> value of 64.42 and 14.60 μg/mL for EO and 0.961, >1 mg/mL for the AE of <i>A. lerchiana</i> and <i>A. santonicum</i>, respectively. The low inhibition on the mycelium growth of studied phytopathogenic fungi was established by applying 2 µL of EO and 15 µL of 100 mg/mL of AE, with the exception of <i>A. lerchiana</i> AE against <i>Botrytis cinerea</i>. These results show that the studied EO and AE exhibited strong phytotoxic and AChE inhibitory activities, providing new data for these species.
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