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The prospect of replacing traditional chemical fertilization with organic and microorganism-based fertilization meets the current demand for more sustainable cropping systems and healthy food. In this respect, research was carried out to evaluate the effects of the factorial combination between four basil cultivars ('Aromat de Buzau', 'Macedon', 'Cuisoare' and 'Serafim') and three types of fertilization, namely chemical fertilization (with a solid chemical fertilizer), organic fertilization (with chicken manure formulate) and microorganisms' fertilization (with microorganisms formulate), on basil yield, biochemical and physiological parameters and essential oil composition. The results showed that the biometric parameters (plant height, number of stems and leaves and leaf area) were significantly influenced by the cultivar; 'Macedon' obtained the highest values of plant height (64.7 cm) and number of stems (20.33) and leaves (618.3) and 'Serafim' the largest leaf area (4901.7 cm2 per plant), while the type of fertilization did not affect these parameters. Regarding the biomass, the influence of the cultivar was not significant on fresh biomass but was significant on dry biomass, with 'Macedon' showing the highest value (56.4 g·plant−1 dry biomass). The mentioned parameters were significantly influenced by the type of fertilization, with the highest values recorded with chemical fertilization. Both the cultivar and the fertilization type significantly influenced the physiological parameters (the total content of assimilatory pigments and photosynthesis). Five phenolic compounds were quantified from leaf extracts by HPLC-MS (caffeic acid, hyperoside, isoquercitrin, rutin and quercitrin). Hyperoside was identified only in 'Macedon', while the rest of the compounds were found in all the cultivars and varied depending on the cultivar and fertilization type. Regarding the composition of the essential oil, variation was found depending on the cultivar and fertilization type. In 'Aromat de Buzau', the main compounds were methyl chavicol and β-linalool; in 'Macedon', geranial and neral; and in 'Cuisoare' and 'Serafim', β-linalool. Moreover, the PCA showed that the 'Serafim' cultivar has exclusive properties compared to the other cultivars. Our results highlight that identifying the most effective interaction between genotype and fertilization type allows to optimize yield and quality targets for sweet basil. [ABSTRACT FROM AUTHOR]