The objective of this study was to identify the chemical composition and antimicrobial activity of acetone extracts of Pinus halepensis Mill (needles and bark) and Olea europaea L. (leaves and bark) obtained from three different altitudes in AL-Jabal AL-Akhder, Libya. The chemical compositions were analyzed by GC/MS. The analyses of the extracts from needles of P. halepensis led to the identification of 38, 15 and 18 different components, representing 93.34, 100 and 97.00% of the total extract at three different altitudes (125, 391 and 851 respectively). However, the extracts from bark contained 6, 4 and 8 compounds, representing 100% of the total extract at three different altitudes, respectively. For the leaf extracts from O. europea, 5, 8 and 10 compounds were identified, representing 100% of the total extracts at three different altitudes (125, 391 and 851 respectively). However, the crud extracts from bark led to the identification of 13, 16 and 15 compounds, representing 97.55, 99.70 and 98.05% of the total extract at three different altitudes, respectively. The chemical classes of the detected compounds confirmed that these extracts contained a complex mixture consisting of sugars, monoterpene hydrocarbons, sesquiterpenes, diterpenes, diterpenoids, terpenophenolics, triterpenes, tetraterpenoids, phenylethanoids, steroids, resins and phthalates. Four pathogenic bacteria and one fungal strain were used to determine the antimicrobial activity. The extracts exhibited antibacterial potency with varying degrees of inhibition with MIC values ranging from 480 to 1300mg/L and the best MICs values observed were 525, 530, 410 and 645mg/L against the growth of Agrobacterium tumefaciens, Erwinia carotovora, Corynebacterium fascians and Pseudomonas solanacearum, respectively for the extract from O. europaea at altitude I. The extracts all extracts exhibited a significant antifungal potency against Botrytis cinerea with varying degrees of inhibition of growth with EC 50 values ranging from 71.10 to 154.71mg/L.