Background - The sixth anthropocene mass extinction in the last 540 million years indicates that human activities including illegal trade is the fundamental threat to the endangered species, their habitats and to the global biodiversity as well. In this context, it is paramount keeping in view the role of hotspot regions in the conservation of biodiversity and their varied potential of ethnobotanical services.The present study quantified the traditional ethnobotanical usage and people’s choices of plants and evaluated the composition, distribution, trade and conservation of plants in Ladakh region, India.Methods - During 2018-2019, ethnobotanical documentation of the plants growing in Ladakh (a part of trans-Himalaya), India, was carried out to collect information regarding different usages of plant species growing in the region through open and close-ended questionnaire surveys and interviews. Multivariate ecological community analysis was used to find the relationship between ethnobotanical usage and plant species.Results - Floristically, a total of 103 species belonging to 80 genera and 38 families were reported to be used as medicine, food, fodder, fuelwood, fragrance, dye, flavor and oil. Asteraceae was the largest family with 20% species, followed by Ranunculaceae (10% species) and Fabaceae (7% species). Three plant usage clusters were determined at a vertical distance value of 0.4 where the clusters are distinctly separated. Preference analysis showed the highest priority of local people for medicinal use of plants followed by fodder and fuelwood. The most frequently used plant parts were leaves followed by roots and flowers. Local Amchis and elderly people had great familiarity with medicinal plants and their ethnic usage. The plant species harvested were sought for self-consumption (65%) and for commercial purposes (35%).Conclusions - The present study provides unique information regarding the ethnobotanical knowledge in Ladakh region and baseline data for future researchers, policymakers, public, land managers, and the other stakeholders to develop scientifically-informed strategies for conservation of natural resources and sustainable use of plant diversity in hotspot regions like trans-Himalayas and other similar biodiversity-rich sites.