Increasing demands by consumers for fresh, nutritional, and convenient food has led to the increase of fresh-cut produce market. Nowadays, there is a turn towards the investigation of natural products (i.e., essential oils, organic acids, and edible coatings) in an effort to lower the usage of chemical synthetic compounds (i.e., chlorine) as postharvest sanitizers. The aim of the present study was to assess the effectiveness of Origanum majorana essential oil (EO), ascorbic acid (AA), chitosan, and their combinations on quality attributes of fresh-cut lettuce stored for six days at 7 °C. When applied, Chitosan+AA resulted to a less acceptable product (visual quality and aroma), while the application of marjoram EO was able to preserve the visual quality of fresh-cut lettuce and at the same time resulted in a pleasant aroma. The application of EO+AA and Chitosan+AA increased total phenolics and antioxidant levels of fresh-cut lettuce on the fourth and sixth day of storage. The EO and EO+AA increased damage index (hydrogen peroxide and lipid peroxidation) of fresh-cut lettuce, while at the same time these treatments decreased the activity of enzymes related with plant tissue browning (i.e., peroxidase activity and polyphenol oxidase). Chitosan decreased total valuable counts and yeasts and molds counts on the sixth day of storage, while EO, AA, EO+Chitosan, and Chitosan+AA decreased yeasts and molds after four days of application. The findings of the present work indicating that the combination of marjoram EO, AA, and chitosan could be considered further as alternative means for fresh-cut produce preservation.