The Jalawastu Cultural Village Community is located in Brebes Regency, Central Java, Indonesia. They practice local traditions and culture through the use of various plants, including medicinal types. Therefore, this research aims to determine the medicinal plant species used with their benefits, cultural importance, and knowledge transfer through interviews and field surveys. These plants were analyzed descriptively, while the cultural importance index was determined quantitatively by calculating the ICS (Index of Cultural Significance) value. The results showed that there were 74 medicinal plants from 37 families. Zingiberaceae was the most widely used at 15.3%, followed by Euphorbiaceae, Fabaceae, Malvaceae, Myrtaceae, and Poaceae at 5.5%. These species could treat 49 disease, and the ICS analysis ranged from 1.5 to 51, where coconut has the highest value at 51. The source of knowledge is obtained from parents, relatives, or neighbors. The intensity of its benefits in the younger generation tends to be low, which can cause the gradual loss of knowledge. In conclusion, the community in the Jalawastu Cultural Village still traditionally uses plants to treat various diseases and provide health care. Meanwhile, medicinal plants in this area have an essential role in the community's lives. It is necessary to carry out scientific research, preclinical and clinical tests to ensure the content of phytochemical compounds, effectiveness, safety, chronic toxicity, pharmacological studies, standardization, and interactions with medicinal ingredients.