This research evaluated the effects of dual-frequency ultrasound-assisted thawing (UAT) on the thawing time, physicochemical quality, water-holding capacity (WHC), microstructure, and moisture migration and distribution of large yellow croaker. Water thawing (WT), refrigerated thawing (RT), and UAT (single-frequency: 28 kHz (SUAT-28), single-frequency: 40 kHz (SUAT-40), dual-frequency: 28 kHz and 40 kHz (DUAT-28/40)) were used in the current research. Among them, the DUAT-28/40 treatment had the shortest thawing time, and ultrasound significantly improved the thawing rate. It also retained a better performance from the samples, such as color, texture, water-holding capacity and water distribution, and inhibited disruption of the microstructure. In addition, a quality property analysis showed that the pH, total volatile basic nitrogen (TVB-N), and K value were the most desirable under the DUAT-28/40 treatment, as well as this being best for the flavor of the samples. Therefore, DUAT-28/40 treatment could be a possible thawing method because it improves the thawing rate and maintains the quality properties of large yellow croaker.