Abstract: In order to evaluate the effect of pulsed light (PL) treatment on the quality changes and microbial diversity of strawberries during storage, strawberry samples were treated with different light intensities and pulse frequence of PL1(100 J, 60 pulse), PL2(300 J, 60 pulse), PL3(500 J, 60 pulse). The changes of decay rate, hardness, color, soluble solids, total phenols content, and anthocyanin content of strawberries were determined during storage at 4±1 ℃. The microbial diversity of the strawberry samples was analyzed using Illumina Miseq PE300 high-throughput sequencing technology. At the end of storage (14 d), the physicochemical indicators showed that the decay rate of strawberries treated with PL2 and PL3 decreased from 59.5% in the control group to 40.75% (PL2) and 16.31% (PL3), respectively. The hardness was significantly higher than that of the control group by 54% and 46%, respectively. It was also observed that PL treatment effectively slowed down the decline of the contents of soluble solids, anthocyanin and total phenols in strawberry. The results of high-throughput sequencing showed that the dominant genera of spoilage microorganisms in strawberry were Dendrobacter spp. and Owenella spp., with the initial relative abundance of 70.14% and 48.38% respectively. PL3 treatment decreased the initial microorganisms significantly, resulting the relative abundance of 37.08% and 2.93%. After 14-day storage, the relative abundance of these two species increased to 82.57% and 73.52%, 65.68% and 42.49%, respectively in control and PL3 treated groups. The results showed that PL could delay the spoilage of strawberries and significantly prolong the storage period of strawberries under refrigeration conditions.