Early blight of tomato is one of the highly destructive diseases caused by an airborne soil-inhabiting fungus Alternaria solani (Ellis and Martin). Seven A. solani strains were isolated from the naturally infected different tomato plants growing in fields and tunnels at Lahore, Kasur, Faislabad, Islamabad, Lalyani Peshawer and vegetable market Lahore, Pakistan. All strains of A. solani showed variations in morphological feature. These fungal strains were identified and compared on the basis of morphological and ITS sequence analysis. All strains shared 87.7% homology with genetic distance of 0.174. Pathogenicity test of the isolated strains confirmed their host specificity and re-isolation of the pathogen from symptomatic tomato fruit fulfilled Koch’s postulates.