Tomato (Lycopersicon esculentum Mill., Family: Solanaceae), a most popular vegetable crop plant propagated by seed as well as seedling. It is cultivated in both tropical and subtropical regions of India. In South Gujarat condition tomato is the important vegetable crop. Wilt of tomato caused by Fusarium oxysporum f.sp. lycopersici (Sacc.) Synder and Hans., it is a serious problem in South Gujarat which damaged the most of the important varieties. Considering the seriousness of the disease, present investigation was carried out. The pathogenicity of ten isolates collected from different locations of South Gujarat from different varieties was proved by the root dip inoculation technique. The cultures were also tallied by cultural characters and microscopic observations. On the basis of pathogenicity, cultural and morphological characters, the pathogen identified as Fusarium oxysporum f.sp. lycopersici (Sacc.) Synder & Hans. These ten isolates were designated, as SGFOL (South Gujarat Fusarium oxysporum f.sp. lycopersici, SGFOL-1 to SGFOL-10). There was a good deal of variation in pathogenic, cultural, morphological and molecular level within the isolates of the pathogen.