The fate and toxicity of silver nanoparticles (AgNPs) and ions in water bodies is largely determined by the natural organic matter (NOM)-mediated redox cycling. However, the process of NOM-mediated redox cycling in the day/night cycles remains elusive. In this study, the inter-transformation between AgNPs and Ag+ ion caused by humic acid (HA) was investigated under controlled light and dark conditions. It was shown that HA induced the reduction of Ag+ into AgNPs in simulated sunlight, and also oxidize AgNPs to release Ag+ in darkness. Kinetics data demonstrated that the rates of AgNPs formation and dissolution increased along with the increment of HA concentrations. Along with the pH increase, the reduction of Ag+ accelerated, but the oxidative dissolution of AgNPs was inhibited. In day-night cycles, the AgNPs and Ag+ concentrations exhibited similar wave-shaped change curves. The peaks of AgNPs and Ag+ ion appeared at 7 p.m. and 7 a.m., respectively. The toxicity of AgNPs/Ag+ to Escherichia coli was determined primarily by the concentration of dissolved Ag+, but also affected by the particle-specific toxicity. The dual role of HA implied that previous reports about the photo-reduction of Ag+ to AgNPs by NOM should be reconsidered, and the oxidizability of HA in darkness strongly affect the transformation and toxicity of AgNPs in water. © 2020. Springer Science+Business Media, LLC, part of Springer Nature.