Nanoplastics (NPs) have become a new type of pollutant of high concern that is ubiquitous in aqueous environments. However, the transport and transformation of NPs in natural waters are not yet fully understood. In this study, the aggregation and photooxidation of NPs were assessed with nanosized polystyrene (PS) as an example, and the effects of dissolved organic matter (DOM) were investigated with Suwannee River fulvic acid (SRFA) as representative DOM. The results showed that simulated sunlight irradiation exhibited negligible effects on the aggregation of PS, while SRFA enhanced its heteroaggregation through hydrophobic interactions. In SRFA solutions, photooxidation of PS with a particle size of 200 nm was observed, which led to an increase in the O/C ratio on its surface at a rate of (2.20 ± 0.40) × 10-2 h-1. This indicates the promotional effect of SRFA on the oxidation of nanosized PS, which is attributed to the generation of the excited triplet state (3SRFA*), hydroxyl radicals (•OH), and singlet oxygen (1O2). Among these reactive species, 1O2 played a crucial role in the oxidation of PS. The findings in this study are helpful for an in-depth understanding of the environmental behavior of NPs in natural waters.