The quality changes, dynamic changes in microbial composition, and diversity changes in large yellow croaker (Larimichthys crocea) during 4 °C refrigeration were studied using 16S rDNA high-throughput sequencing technology, and the total viable count (TVC), total volatile basic nitrogen (TVB-N), and thiobarbituric acid-reactive substances (TBARS) were determined. The results revealed a consistent increase in TVC, TVB-N, and TBARS levels over time. On the 9th day, TVC reached 7.43 lg/(CFU/g), while on the 15th day, TVB-N exceeded the upper limit for acceptable quality, reaching 42.56 mg/100 g. Based on the 16S rDNA sequencing results, we categorized the storage period into three phases: early storage (0th and 3rd days), middle storage (6th day), and late storage (9th, 12th, and 15th days). As the storage time increased, both the species richness and diversity exhibited a declining trend. The dominant genus identified among the spoilage bacteria in refrigerated large yellow croaker was Pseudomonas, accounting for a high relative abundance of 82.33%. A comparison was carried out of the spoilage-causing ability of three strains of Pseudomonas screened and isolated from the fish at the end of storage, and they were ranked as follows, from strongest to weakest: P. fluorescen, P. lundensis, and P. psychrophila. This study will provide a theoretical basis for extending the shelf life of large yellow croaker.