Carbon dots (CDs) obtained from rapeseed pollen with a high production yield, good biocompatibility, good water solubility, low cost, and simple synthesis are systematically characterized. They can be directly added to Hoagland nutrient solution for planting hydroponically cultivated Lactuca sativa L. to explore their influence on the plants at different concentrations. By measuring lettuce indices of growth, morphology, nutrition quality, gas exchange, and content of photosynthetic pigment, amazing growth-promotion effects of CDs were discovered, and the mechanism was analyzed. Moreover, the in vivo transport route of CDs in lettuce was evaluated by macroscopic and microscopic observations under UV light excitation. The results demonstrate that pollen-derived CDs can be potentially used as a miraculous fertilizer for agricultural applications and as a great in vivo plant bioimaging probe.