We aimed to evaluate the bioherbicide potential of Eucalyptus saligna leaf litter essential oil, its fractions and major compounds. Six essential oil fractions were obtained by preparative thin‐layer chromatography and analyzed by gas chromatography/mass spectrometry. Effects of the oil, its fractions and major compounds (1,8‐cineole and/or α‐pinene) diluted in aqueous solution were assessed on germination and seedling growth. Recipient species were Lactuca sativa (model), Amaranthus viridis (weed), Eragrostis plana (weed), and Paspalum notatum (forage). The essential oil was more phytotoxic on A. viridis and L. sativa, followed by E. plana, and caused no effects on P. notatum. Amaranthus viridis was inhibited by all treatments, mainly the essential oil, α‐pinene and fraction 6, whereas E. plana was more affected by the oil fractions. Results revealed the species‐specific phytotoxic effects of E. saligna essential oil, indicating its potential use for controlling A. viridis and E. plana. Phytotoxic effects of essential oils or their components should not be generalized, as effects may change according to recipient species.