Ethnomedicinal knowledge is a valuable component of traditional health care systems and plays an important role in the discovery of novel therapeutic agents. The present study was carried out with an aim to document the medicinal plant diversity and associated indigenous knowledge in Karauli district of Rajasthan, India, where rural and tribal people still depend on herbal remedies for their primary healthcare. The authors have conducted extensive field surveys in different villages and ethnomedicinal information has been collected through semi-structured questionnaires by personal interviews and discussion with traditional healers, elderly villagers and knowledgeable local informants. The plant specimens were identified with the help of standard taxonomic literature and information on botanical name, family, ailments treated and methods of herbal preparation was recorded systematically. The survey recorded a total of 94 species of medicinal plants belonging to 43 families. Fabaceae was the dominant family followed by Malvaceae, Asteraceae, Lamiaceae, Poaceae, Moraceae, Euphorbiaceae and Amaranthaceae. The medicinal plants documented are used traditionally for the treatment of a wide range of ailments like skin diseases, wounds, gastrointestinal disorders, respiratory infections, diabetes, fever, rheumatism, arthritis, urinary disorders, liver diseases and inflammatory conditions. Preparation by decoction was the most frequently used method, followed by paste, powder, juice, oil, and smoke inhalation. Multiple therapeutic applications of several plant species like Azadirachta indica, Ocimum sanctum, Tridax procumbens, Syzygium cumini, Calotropis procera and Holoptelea integrifolia indicated their importance in indigenous healthcare system. The results indicate that Karauli district is a rich reservoir of ethnomedicinal plant diversity and possess rich traditional knowledge which is still used to fulfill the health care needs of local communities. The documented information provides an important baseline for future phytochemical, pharmacological and clinical investigations and emphasizes the dire need for conservation of medicinal plant resources and preservation of indigenous knowledge before it is lost irretrievably.