The objective of this study was to develop a simple anther culture protocol for a range of Saudi wheat genotypes. Seven wheat genotypes were evaluated in anther culture on five different medium protocols for their ability to initiate callus and green plants. The estimates of significance for the effects of genotypes, and medium protocols used, and their interactions on callus induction, callus weight and shoot formation derived from anther explants indicated that the in vitro traits were significantly influenced by the genotypes, medium protocols, and their interactions. The percentage of explants that developed calli ranged from 0.41% (Lang) to 15.39% (Irena) averaged across the five medium protocols with an average 4.45%. The genotype Irena produced the highest average means of shoot formation (69.65%) across medium protocols. The genotype Yecora Rojo (13.73%) was significantly inferior to all other tested genotypes for shoot formation.