Androgenesis is the newest and most secure method to obtain haploid plants in vitro, where vegetative or generative nucleus of a pollen grain is stimulated to develop into a haploid individual. There are different methods for regeneration and formation of microspores in various genotypes of tobacco. In this case, the level of androgenesis was investigated in three tobacco genotypes. NN-medium was used as a basic medium for microspores development and MS-medium for rhizogenesis and organogenesis, together with adequate combinations of plant hormones (JAA, BAP, adenine, glutamine and kinetine).