Phytophthora species are particularly aggressive plant pathogens and are often associated with the decline of many tree species, including oak and beech. Several fungi and bacteria species are known as potential antagonists usable as biological control agents. Phosphonate (H3PO3), commonly branded as phosphite, has also been used in the past years to protect trees against invasive Phytophthora spp.. This study aimed at comparing the effects of selected antagonist microorganisms and phosphonate, when applied by microinjection or leaf treatment. Antagonistic species were first selected for their high inhibitory activity against problematic Phytophthora species, such as Phytophthora cactorum, P. quercina and P. plurivora attacking Quercus robur and Fagus sylvatica in Polish forests. Three endophytic species Trichoderma atroviride (two strains), T. harzianum and Bacillus amyloliquefaciens showed a high control activity, and their efficacy was then assessed in comparison with a phosphonate treatment. Two application methods were experimented in this study: injection of a solution of spores or phosphonate into the sap vessels of beech or a foliar treatment on oak. Phosphonate and two strains of Trichoderma significantly reduced the necrotic area on oak leaves inoculated with P. plurivora and one strain of T. atroviride significantly reduced necrotic areas on beech branches. These results are therefore promising of a novel way to control Phytophthora spp. in forest stands and nurseries.