In the present study, a protocol of isolated microspore culture was optimized to regenerate green plants in Turkish durum wheat genotypes (Kiziltan-91, C-1252, Mirzabey 2000 & Kunduru-1149). The bread wheat cultivar (Gun-91) was used as control because of its high androgenic response in the microspore culture. First significant step was to treat the anthers with four pretreatments (cold, cold with mannitol, cold with sorbitol & mannitol at room temperature). Anther maceration was used as an isolation method and microspores were plated on induction culture medium supplemented with arabinogalactan-proteins (AGP) and ovary coculture. When the embryos reached the size of 2 mm, they were transferred to the differentiation medium having a combination of phenylacetic acid (PAA) and gibberellic acid (GA(3)). The best results were obtained with the pretreatment of mannitol (+4 degrees C) for 7 d on providing embryos and regenerated green plants in four durum wheat genotypes. The cultivar Kiziltan-91 gave the best response for embryo (>2 mm) formation (20.63%) and cultivar Kunduru-1149 for green and total plant regeneration (6.25% and 18.75%). These results indicated that the present protocol performed well to increase the embryo formation and green plant regeneration in the recalcitrant durum wheat genotypes studied.