A laboratory study was conducted to evaluate the essential oil composition, total phenolic compound, total flavonoid compound and the antioxidant activity in seed and callus induced from tissue culture of two fennel genotypes. The influence of genotype, explants type and growth regulators concentrations on callus induction was investigated. Results strongly showed that there were significant differences between two fennel genotypes. Generally the highest values of all parameters were obtained from Balady genotype. Callus was induced from two explants type on MS medium containing different combinations of 2,4-D and Kinetin. The essential oil characterization by GC revealed that seed oil had the highest amount of trans-Anethol of two genotypes. On the other hand, callus oil from Balady genotype had the highest amount of α-Pinene while Romanesco genotype had the highest amount of p-Cymene. Balady genotype had the highest amount of total phenolic and flavonoid content in seed as well as callus induced from tissue culture compared with Romanesco genotype. The antioxidant activities of both genotypes were evaluated and Balady genotype showed the highest DPPH_ scavenging activity expressed as IC50 compared with Romanesco genotype.