The aim of this study was to assess the effect of chitosan/zein-based coating containing free and nano-encapsulated Pulicaria gnaphalodes (Vent.) Boiss. aqueous extract (PGE) on microbial, chemical, and sensorial attributes of rainbow trout during refrigerated storage. Different concentrations of soy-lecithin: cholesterol (60:0, 50:10, 40:20, and 30:30) were used to prepare nanoliposomes with the mean size range of 142–166 nm according to the thin-film hydration-sonication method. Results demonstrated the size distribution and encapsulation efficiency (EE) of 0.81–0.92 and 49.88–73.05, respectively. Nanoliposomes with the lowest droplet size and highest EE were selected for incorporating in the film samples. Information on the Fourier transform infrared (FT-IR) spectrum showed some new interactions between chitosan–zein (Cs-Zn) and nanoliposomes. Biological evaluation results showed that chitosan-zein coating could retard spoilage of rainbow trout fish meat as seen in lower thiobarbituric acid reactive substances and peroxide value index value (p < .05). Nanoliposomes of Pulicaria gnaphalodes (Vent.) Boiss. extract resulted in the controlled release of the bioactive agents on the surface of fish, which prolonged antimicrobial and antioxidant activity of coating during 14 days compared with other groups. Cs-Zn coating with PGE-loaded nanoliposomes can be considered as a potential active packaging in the meat industry.