Somatic hybrid plants between N. tabacum (IVB ( bar )) and N. megalosiphon ( npt II) were recovered after polyethylene glycol (PEG) - mediated fusion and electrofusion. The hybrid calli were selected on plant regeneration medium on the basis of dual resistance to bialaphos and kanamycin or UV inactivation of the donor species protoplasts. The hybrid nature of the clones obtained was confirmed by AFLP analysis. According to their morphology the plants regenerated were classified in the following groups: 1) Self fertile hybrid plants of N. tabacum phenotype; 2) Self fertile hybrid plants of N. megalosiphon phenotype; 3) Self sterile plants with N. tabacum habit, leaf morphology and flowers closer to N. megalosiphon ; 4) Self sterile plants with N. megalosiphon habit, abnormal leaves and flower morphology closer to N. megalosiphon ; 5) Self sterile plants of N. megalosiphon type with abnormal characters which were obtained after UV irradiation of the donor protoplasts. Pollen staining with acetocarmine and pollen germination study of the self sterile plants revealed viable pollen formation in the hybrid plants from the third group. The DNA content of the regenerated plants was determined by flow cytometry. The results provided evidence for non preferential loss of one of the parental genomes and spontaneous asymmetrization of the hybrid plants. The results from the present study show that by means of somatic hybridization the crossability barriers between distantly related species can be overcome and greater genetic diversity in the hybrid clones can be achieved. This technique can supplement or substitute the conventional plant breeding methods.