This work presents the biocidal effects and chemical compositions of two essential oils (EOs) obtained by hydrodistillation of Eucalyptus flocktoniae subsp. flocktoniae and E. flocktoniae subsp. hebes. The two subspecies studied had different chemical composition, when analysed by gas chromatography coupled to mass spectrometry with 1,8-cineole (56.98%), trans-pinocarveol (20.38%) and α-pinene (5.86%) being the major components of E. flocktoniae subsp. flocktoniae and spathulenol (25.09%), p-cymene (21.20%), 1,8-cineole (10.74%) and α-pinene (8.93%) are the major components of E. flocktoniae subsp. hebes. These oils were evaluated for their insect antifeedant, nematicidal and phytotoxic activities. The biocidal tests showed that E. flocktoniae subsp. hebes was the most active against Myzus persicae and Rhopalosiphum padi. While, E. flocktoniae subsp. flocktoniae was not antifeedant. None of the investigated EOs were active against both Spodoptera littoralis and Meloidogyne javanica root-knot nematode. Moreover, the EOs extracted from these two subspecies showed a significant phytotoxic effect.