This study was conducted to determine the effects of genotype, medium and 2,4-D concentration on the anther culture of wheat (Triticum aestivum L). In the study, one cultivars, three lines and six F1 progenies produced from the crosses of these cultivars and lines used as donor plants. The anthers were cultured on two different media (P-II and mMS) containing different 2,4-D concentrations (4.5 muM, 6.8 muM, 9 muM and 11.3 muM). The results of the study showed that frequency of responding anthers was significantly influenced by the genotypes and 2,4-D concentrations. The mean frequency of responding anthers varied from 15.0 % to 0.6 % among the genotypes. 4.5 muM 2,4-D concentration produced the highest frequency of responding anthers (61 %), and 68 muM 2,4-D the lowest one (3.5 %). The effects of media on the frequency of green plantlets/100 anthers changed as depending upon the genotypes. The cultivars Seri-82 giving 11.6 green plantlets per 100 anthers was superior to other genotypes tested in that character. P-H Medium with 9 muM 2,4-D produced higher mean number of green plantlets per 100 anthers than did P-H media with the other concentrations of 2,4-D, while 6 8 muM 2,4-D for mMS medium was optimum concentration of 2, 4-D for an optimum number of green plantlets per 100 anthers.