Bacterial infections pose a serious risk to human health. Although antibiotics have been extensively used to combat such infections, their widespread and indiscriminate use has led to the emergence of an increasing number of multidrug-resistant strains, emphasizing the urgent necessity for safe and effective new antimicrobial agents. Therefore, in this study, carbon dots (CDs) were synthesized from corn silk, a natural antimicrobial material sourced from the medicinal food homologous Chinese Medicine, and their structural characteristics and antimicrobial efficacy were analyzed. The experimental results indicate that the corn silk carbon dots (CS-CDs) retain the antimicrobial active groups (-COOH and -CN) present in corn silk and display potent antimicrobial activity against Staphylococcus aureus, Escherichia coli, and Candida albicans and have low toxicity and good biocompatibility, indicating significant clinical potential. Consequently, this study has advanced the development of CDs with robust antimicrobial capabilities derived from corn silk, thereby demonstrating the promising potential of CS-CDs as effective antimicrobial nanomaterials. This research lays the groundwork for the innovative reutilization of existing herbal resources through modern nanomaterial synthesis techniques.