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Practical Application Fermented walnut meal (FW) has antifungal activity against Penicillium victoriae, a fungus responsible for Rosa roxbughii Tratt spoilage. This study characterized and applied ultrasonic‐assisted antifungal film loaded with FW to preserve R. roxbughii Tratt during near‐freezing temperature (NFT). Results showed that O2 and CO2 transmission rates decreased by 80.02% and 29.05%, respectively, and antimicrobial properties were improved with ultrasound at 560 W for 5 min and 1% FW. Fourier transform infrared spectroscopy and X‐ray diffraction results revealed ultrasound improved hydrogen bonds and inductive effect via ─NH, ─OH, and C═O bonds. The addition of FW led to the formation of CMCS‐GL‐FW polymer via C═O bond. Thermogravimetric analysis and transmission electron microscope results demonstrated thermal degradation process was decomposed by ultrasound, and the internal structure of P. victoriae was accelerated by the addition of FW. Compared to the U‐CMCS/GL group, the vitamin C content, peroxidase, and catalase activities of U‐CMCS/GL/FW were enhanced by 4.24%, 8.52%, and 14.3% during NFT (−0.8 to −0.4°C), respectively. Particularly, the fungal count of the U‐CMCS/GL/FW group did not exceed 105 CFU g−1 at the end of storage, and the relative abundance of P. victoriae decreased to 0.007%. Our findings provide an effective route for agricultural waste as natural antifungal compounds in the active packaging industry.In this study, the barrier and antimicrobial properties of film were successfully improved by ultrasonic treatment and loaded fermented walnut meal. The ultrasonic‐assisted antifungal film loaded with fermented walnut meal effectively delayed the degradation of nutrients and reduced microbial invasion of Rosa roxburghii Tratt. These results provide a theoretical basis for the application of agricultural waste in the food packaging industry. [ABSTRACT FROM AUTHOR]