Withania (Family: Solanaceae) is a highly acclaimed genus in the Indian Ayurvedic system of medicine. In Ayurveda, Withania is known to promote physical and mental health and used to treat almost all the disorders that affect human health. Withania somnifera andWithania coagulans are the two most esteemed species of this genus having high medicinal significance. These species are natural source of withanolides (steroidal lactones) which are used as ingredients in many formulations prescribed for a variety of diseases. Many pharmacological studies have been conducted to investigate the properties of Withania as a multi-purpose medicinal agent. Advances in biotechnology, especially in vitro culture techniques, molecular biology and metabolite profiling provided new insights for conservation and management of plant genetic resources and better harvesting of drugs from medicinal plants. This review presents a consolidated account of the phytochemistry, pharmacology and biotechnology involving in vitro propagation, genetic transformation and metabolite profiling in W. somnifera and W. coagulans. Key words: Withania, phytochemistry, pharmacology, withanolides, micropropagation, metabolite profiling.