To elucidate the taxonomic status of Ruppia drepanensis Tineo ex Guss. (Alismatales, Ruppiaceae), we performed morphological analysis and DNA barcoding of historical materials (including the lectotype) and fresh samples (including those from a recently discovered population near the locus classicus in Sicily, Italy). We conclude that R. drepanensis is a separate species, closely related to R. spiralis L. ex Dumort., that occurs in temporary inland waters from the western to central sectors of the Mediterranean region. We also highlight the importance of vouchers and the need to link molecular investigations to field, ecological, and morphological investigations.