The inefficient delivery of herbicides causes unpleasant side effects on the ecological environment. Protoporphyrinogen oxidase (PPO)-inhibiting herbicides rely on the presence of external light to exert the activities and thus their performance in the field is extremely susceptible to the light environment. Here, taking acifluorfen (ACI) as a model PPO-inhibiting herbicide to enhance efficacy by boosting the utilization rate of sunlight, amphiphilic cationic CDs (CPC-CDs) from cetylpyridinium chloride (CPC) as a precursor, is first prepared as a supplementary light source generator, and subsequently co-assembled with ACI through non-covalent bond interactions to obtain the stable fluorescent nanoparticles (ACI@CPC-CDs). ACI@CPC-CDs with fascinating physicochemical properties can penetrate the leaves of weeds through the stomata and undergo a long-distance transport in the cell intervals. Under low light intensity, CPC-CDs can be applied as the internal light source to promote the formation of more singlet oxygen to damage the leaf cell membrane, consequently improving the herbicidal activity of ACI. Moreover, the safety evaluation of ACI@CPC-CDs demonstrates no risk to non-target organisms and the environment. Therefore, this work offers a promising strategy for the efficient delivery of light-dependent PPO-inhibiting herbicides and opens new insights into the application of CDs in the development of sustainable agriculture.