Tobacco (Nicotiana tabacum L. and N. rustica L.) is an important crop for the economy of India. Gujarat is a leading tobacco producing state of India contributing 33 % of total tobacco production from 0.45 lakh ha. A large number of tobacco varieties have been developed during last six decades for quality and quantity improvement throughout the world using local selection, mutation breeding, hybridization and exotic introduction followed by pedigree selection. However, no systemic effort has been made to understand the existing diversity pattern in tobacco cultivars of Gujarat, which is crucial to define future breeding strategy in tobacco improvement.