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Arbuscular mycorrhizal fungi (AMF) and elevated CO 2 (eCO 2 ) have been effectively integrated to the agricultural procedures as an ecofriendly approach to support the production and quality of plants. However, less attention has been given to the synchronous application of AMF and eCO 2 and how that could affect the global plant metabolism. This study was conducted to investigate the effects of AMF and eCO 2 , individually or in combination, on growth, photosynthesis, metabolism and the functional food value of Thymus vulgare . Results revealed that both AMF and eCO 2 treatments improved the photosynthesis and biomass production, however much more positive impact was obtained by their synchronous application. Moreover, the levels of the majority of the detected sugars, organic acids, amino acids, unsaturated fatty acids, volatile compounds, phenolic acids and flavonoids were further improved as a result of the synergistic action of AMF and eCO 2 , as compared to the individual treatments. Overall, this study clearly shows that co-application of AMF and eCO 2 induces a synergistic biofertilization impact and enhances the functional food value of T. vulgare by affecting its global metabolism.