Many isogenic dihaploid lines from Virginia type tobacco were obtained using haploidy by androgenesis method. These lines were studied for their physiological characteristics, industrial quality, chemical content and behaviour relative to diseases caused by TMV, PVY, PVQ and Phytophthora parasitica. With regard to their physiological characteristics, all lines were classified into mid-early and mid-late varieties. Grading was found satisfactory, with 80% of dried leaves in superior and first grades. Regarding their chemical content, the lines showed low nicotine (1%) and high soluble carbohydrates content (25% on an average). Some of the obtained lines are of interest for their resistance to the studied diseases. Yield and nicotine content were generally lower for these isogenic dihaploid lines than for their hybrid parents. These results were confirmed by other authors. Selected lines will be used as initial material for breeding and for production of heterotic F1 hybrids.