This ethnobotanical study was conducted in Ambassel District, South Wollo Zone, Ethiopia, to document the diversity, traditional use, and conservation status of medicinal plants among the local community. A descriptive cross-sectional design was employed, integrating both quantitative and qualitative approaches. Data were collected from 129 informants (including 21 key informants) selected through purposive and random sampling across eight kebeles, using semi-structured interviews, field observations, and questionnaires. The study identified 74 medicinal plant species belonging to 38 families, with Asteraceae and Fabaceae being the most represented. Most medicinal plants (60%) were sourced from wild habitats, while herbs constituted the dominant growth form (42%). Leaves were the most frequently used plant part (47%), and remedies were primarily prepared by crushing or pounding fresh materials and administered orally. Informant consensus was highest for dermal and digestive ailments, indicating strong shared knowledge and effective traditional remedies for these conditions.for these. Multipurpose species remedies such as Vachellia abyssinica and Cordia africana were highly valued but faced significant threats from agricultural expansion, charcoal production, and firewood collection, leading to habitat degradation. Knowledge transmission was mainly oral, predominantly from parents and elders to younger generations, making it vulnerable to erosion due to modernization and youth disinterest. Preference and direct matrix rankings highlighted the importance of species like Allium sativum, Vachellia abyssinica, and Cordia africana in local healthcare and livelihoods. The study concludes that urgent conservation measures are needed, including community awareness, sustainable harvesting, documentation of indigenous knowledge, and integration of traditional medicine into biodiversity conservation policies to safeguard both medicinal plant diversity and cultural heritage in Ambassel District. Recommendations include implementing community-based conservation, promoting awareness, systematic documentation, policy integration, and supporting alternative livelihoods to reduce pressure on wild resources