Context: Ethnobotanical studies are the basis for the acquisition of knowledge about the use of medicinal plants. Aims: To characterize the traditional use of medicinal plants by the population of the municipality of Santa Clara, Cuba. Methods: A field research in nine districts of Santa Clara municipality, Villa Clara province, Cuba was conducted. A questionnaire to key informants from urban and rural localities of both genders, civil status and educational levels, was applied; the sampling was intentional. The Use Value Index and the Significant Use Level were calculated. Results: Ninety-one species of medicinal plants belonging to 27 families were identified; Lamiaceae (13), Compositae (12) and Vervenaceae (6) were the most represented. The most used parts of the plants were leaves (58%) and roots (10%). The most cited preparation technique was the decoction (56.7%). The most frequently cited mixtures were: leaves with flowers in 4% and leaves with roots in 3%. The most treated diseases by the population were respiratory, including bronchial asthma (25.4%), followed by gastrointestinal (23.6%). 19% of the key informants recognized that there were plants contraindicated due to their toxicity. 29.6% of the total species reported showed a significant level of use higher than 20%, so they can be considered important for scientific evaluation and validation. Conclusions: Ninety-one species belonging to 27 families were identified; Lamiaceae, Compositae and Verbenaceae were the most represented. The most used parts of the plants were leaves and roots. The decoction technique was the most cited. The most treated disorders were respiratory and gastrointestinal. 29.6% of the reported species showed significant levels of use greater than 20%.