Background: Hyssopus officinalis L. belongs to Lamiaceae. The herb has a long history of medicinal uses whereas essential oil is widely being used in cosmetics, food ,and pharmaceutical industries worldwide, as well as fresh herb is usually used in cooking. This work was conducted to investigate the effect of Micro-elements on the growth and essential oil of Hyssop (Hyssopus officinalis), cultivated under Egyptian condition. This two-year randomized complete block experiments were performed foliar spraying with Zinc (Zn), Magnesium (Mg), Manganese (Mn), and Iron (Fe) on Hyssopus officinalis L. Results: The study revealed that Zinc and Magnesium had pronounced effects, enhancing plant height, aerial parts, whole plant dry weight, and branching. Moreover, these micro-elements positively impacted flowering and various chemical constituents, including chlorophyll, carotenes, potassium, protein, phosphorus, nitrogen, and total carbohydrates. Foliar application of Zn, Mn, and Fe significantly increased essential oil percentage, resulting in a notable 35% boost compared to the control. The dominant compound in the essential oil composition was trans-3-pinanone (63.37%). Conclusion: Micro-nutrients specially Zinc and Magnesium emerged as key influencers on growth characteristics and positively affected hyssop's chemical composition; chlorophyll A & B, carotenoids, and total carbohydrates with improving essential oil yield and its components that emphasizies the potential of micro-nutrients applications to optimize the cultivation of this valuable herb.