Cheilanthes argentea is a fern belonging to Pteridaceae family which is found mainly in the rock crevices of sub-tropical hilly areas. It has been recognized to possess desiccation tolerant properties because it survives complete desiccation and resurrects when water is available. An extensive field survey was conducted to collect C. argentea from the Nandhi hills State Forest, Chikkaballapura district of Karnataka (Coordinates 13.3702° N, 77.6835° E), the detached fronds obtained from green house maintained plants were hydrated and subsequently desiccated under in-vitro condition. The Relative Water Content decreased in desiccated stage to 4% of its original weight. The fronds regained 96% of its original weight and Relative Water Content during rehydration. The rehydrated fronds largely regained its original morphology. Also, 80% of the total chlorophyll content was retained during desiccation. Further, activities of antioxidant enzymes namely Superoxide dismutase (SOD), Peroxidase (POD), Catalase (CAT) and Glutathione reductase (GR) increased significantly during desiccation. Lipid peroxidation was found to be higher during desiccation when compared to other stages. Proline and sucrose content was higher during desiccation while the concentration of starch decreased due to desiccation. The results obtained in this study demonstrates that C. argentea is potent to tolerate desiccation with its physiological and biochemical defense mechanisms.