Valeriana wallichii, a perennial herb belonging to family Valerianaceae, is an important medicinal herb of the Himalayan region. The incessant exploitation of nature for meeting the demands of the pharmaceutical industry has put unbearable pressure on its natural habitats. A study on its physiological, biochemical, growth and reproductive attributes was planned. Physiological study revealed that ex-situ (outside their natural habitat) populations faced severe stress as compared to in-situ (natural habitat) plants. The difference in the performance of these habitat plants was related to superoxide and H2O2 in the leaves. Photosynthetic attributes were increased in in-situ populations. Proline content and its biosynthetic enzymes ornithine aminotransferase, and pyrroline-5-carboxylate reductase showed an increase in ex-situ plants; proline oxidase decreased. Glucose-6-phosphate dehydrogenase, shikimic acid dehydrogenese, phenylalanine lyase, and flavonoids content showed an increment in ex-situ plants. Antioxidants enzyme superoxide dismutase, catalase, ascorbate peroxidase and reduced glutathione showed an increment in ex-situ conditions. Growth and reproductive attributes were more in ex-situ plants. The observations made are suggestive that a comprehensive conservation programme involving in-situ as well as ex-situ strategies will be effective for the conservation and long term survival of the species.