With the rise of nanomaterials in recent years, a variety of carbon dots (CDs) with good properties synthesized by chemicals have been developed. Herein, Enrofloxacin (ENR) can be detected by environmentally friendly poplar flower CDs (Pf-CDs). Based on the concept of green chemistry, the precursor material is waste poplar flower, which is an easily obtainable plant biomass. Without any special treatment, the whole preparation process for Pf-CDs is quite simple. Meanwhile, the prepared Pf-CDs have outstanding properties such as brilliant luminous performance and good biocompatibility. The Pf-CDs are typically 6.40 nm in size, with excitation/emission wavelengths of 368/445 nm, respectively. It is worth noting that intermolecular hydrogen bonds are formed between Pf-CDs and ENR, leading to the occurrence of aggregation-induced-emission (AIE). The fluorescence signal of Pf-CDs showed a linear relationship with the added ENR concentration of 0.2–100 µM, with a detection limit of 0.076 µM. With a recovery rate of 94.8%-103.15%, it has shown its potentiality and wide application in the detection of ENR in dairy products and domestic water.