Due to the importance of the stage of maturity on several olive pomological parameters and oil quality indices, and to the interest that Tunisian olive oil production has recently received, the optimal harvesting period for the main Tunisian olive cultivar, Chemlali, was assessed. For the first time, a four-season crops study, carried out in three representative geographical areas, was focused on both olive fruit pomological parameters and oil chemical composition at different stages of maturity. The stage of maturity was the factor showing the highest influence on the major part of these parameters, followed by the crop year. To guarantee a reasonable fat content and a good chemical quality of oil, in particular a harmonious acidic composition, acceptable UV absorbance, and a content high of antioxidants, unsaponifiable matter and sterols, the optimal harvesting period for Chemlali olives would appears to be between the end of November to the middle of December, which corresponds to a maturity index between 2.5 and 3.5.