Basil (Ocimum species) represents an extraordinary group of aromatic plants that have gained considerable economic importance, primarily due to their essential oils, which have applications in medicine, culinary, and perfumery. The Ocimum genus encompasses more than 60 species of herbs and shrubs originally native to tropical regions. This genus stands out for its remarkable diversity, displaying a wide spectrum of variations in phenotype, chemical composition, and genetic makeup. In addition to genetic factors, the growth, development, and essential oil production of basil are also influenced by environmental conditions, ontogeny, and various other factors. Consequently, the primary objective of this study was to explore the diversity in both the morphological characteristics and essential oil composition among basil genotypes preserved within the gene bank of the Hungarian University of Agriculture and Life Sciences’ Department of Medicinal and Aromatic Plants. The investigation involved the assessment of fifteen basil genotypes, representing four distinct species: Ocimum basilicum (including ‘Anise’, ‘Clove’, ‘Fino Verde’, ‘Licorice’, ‘Mammoth’, ‘Mrs. Burns’, ‘Thai tömzsi’, ‘Thai hosszú’, and ‘Vietnamese basil’), Ocimum sanctum (green holy basils), Ocimum citrodora (Lemon basil), and Ocimum gratissimum (African and Vana holy basil). The genotypes exhibited significant variations in their morphological growth, essential oil content (EOC), and composition. African basil produced more biomass (408.3 g/plant) and showed robust growth. The sweet basil cultivars clove, licorice, Thai tömzsi, and Thai hosszú also exhibited similar robust growth trends. Vietnamese basil, on the other hand, displayed the lowest fresh biomass of 82.0 g per plant. Both holy basils showed EOC levels below 0.5%, while Mrihani basil stood out with the highest EOC of 1.7%. The predominant constituents of the essential oil among these genotypes comprised estragole, thymol, methyl cinnamate, linalool, and eugenol. In conclusion, this study showed that the genotypes of basil stored in the department’s gene bank exhibit a wide range of variability, both within and between species.