Tobacco (Nicotiana tabacum L.) is one of the most important commercial and industrial crops in the world. Genetic diversity studies provide estimates on the level of genetic variation among diverse materials that can be used in germplasm management, varietal protection, and tobacco improvement. Forty and two tobacco cultivars of three known ecotypes from different origins were genotyped by three mitochondrial-specific markers including two PARSs and one Atp1- sepecific primer pair. The most diversity in terms of Shannon index was observed within Burley ecotype. The largest genetic distance was observed between Virginia and Basma. A low Nei`s distance was obtained between 42 cultivars, indicating that a large proportion of valuable tobacco germplasm may already have been lost through the popularity of certain cultivars in commercial planting and the continuous artificial selection.. Cluster analysis using the unweighted pair group method with arithmetic mean (UPGMA) showed that the cultivars could be grouped into three classes, with no significant relationship between clustering groups and ecotypes.
