A simple one-step hydrothermal green approach was reported for the preparation of carbon dots (CDs) without any further decoration or modification with papaya powder as natural carbon source. In this economical and eco-friendly system, deionized water or 90% ethanol was used as solvent to produce water-soluble or ethanol-soluble CDs, respectively, termed as W-CDs and E-CDs. The quantum yield (QY) for W-CDs was 18.98%, while that for E-CDs was 18.39%. The potentials of the prepared carbon dots toward diverse applications were thoroughly investigated. W-CDs and E-CDs provide promising probes for fluorescence detection of Fe3þ, offering limits of detection of 0.48 μmol L1 and 0.29 μmol L1, respectively. W-CDs was further demonstrated to be a promising probe for fluorescence sensing of Escherichia coli O157: H7, along with a limit of detection of 9.5 104 cfu mL1. Meanwhile, both W-CDs and E-CDs exhibit favorable biocompatibility, and demonstrated to be efficient for Hela cell imaging.