Due to the disadvantages of consuming fat and synthetic preservatives, the demand to reduce them in lipid-based products like mayonnaise is increasing. In the current research, there were two goals, the first one was studying the effect of using oleaster flour in different concentrations (4%, 6%, and 8%) as a natural preservative, whereas the second one was studying the effect of oleaster as different fat replacement (FR) levels (10%, 20%, 30%, and 40%) on the physicochemical, antioxidant, and the rheological properties and stability of reduced-fat mayonnaise samples. Given results showed that with increasing the oleaster concentration, the antioxidant property increased significantly. The peroxide value after 60 days of storage for the 30% FR 8 was 2.01%, compared to the control sample without antioxidant and with TBHQ, which were 10% and 2.68%, respectively. The highest stability index (100%) was observed in the 30% FR and 40% FR samples. In terms of rheological characteristics, the 30% FR 8 oleaster showed the highest viscosity and the lowest frequency dependency. It can be concluded that oleaster has a high potential to be used in the formulation of low-fat mayonnaise as a fat replacer.