This study was carried out to determine the effects of genotype and media on haploid plant formation of 12 pepper genotypes (Capsicum annuum L.) via in vitro anther culture. The buds were cultured in 2 different media (O1 and O2) and, the results revealed that the media had effects on the development of anthers, embryo and plantlet initiation. When the anther developmental status is examined, the lowest value was obtained from the SU-31 genotype with 2.47%, while cv. Flinta F1 produced the highest rate with 12.79%. The highest growth rates were obtained from the O2 medium, and the cv. Dolphin variety produced a remarkable result with 25.0% in this medium. The cultivar Dolphin was the favorable genotypes with 8 embryos and 5 plants, while no embryo and plant were obtained from cv. Flinta F1 and SU-34. A total of 2868 anthers were cultured, 195 anthers were enlarged and the growth rate was 6.8%. Finally, 37 embryos and 18 plants were obtained and the frequencies were 1.29% and 0.63%, respectively. As a result of stomatal observations, 13 plants were detected haploid and the others were double haploid.