Factors affecting the successful rapid proliferation and rooting of male pistachio (Pistacia vera L.) cv. Atlı, were studied. The most suitable type of cytokinin [6 benzyladenine (BA), kinetin (Kin), or thidiazuron (TDZ)], and the effect of eight different concentrations of BA (0.0675, 0.125, 0.25, 0.5, 1.0, 2.0, 4.0, or 8.0 mg l^–1) were evaluated to optimise shoot proliferation. The auxins -naphthaleneacetic acid (NAA), indole-3-acetic acid (IAA), and indole-3-butyric acid (IBA), each at 2.0 mg l^–1, or different concentrations (0.5, 1.0, 2.0, 4.0, or 8.0 mg l^–1) of IBA, and the effect of explant size (1.0, 2.0, 3.0, or 4.0 cm) were assessed for root induction. The highest number of new microshoots per explant (5.64 ± 0.07) was obtained 4 weeks after culturing on Murashige and Skoog (MS) medium supplemented with 1.0 mg l^–1 BA. Again, shoot length was highest in 1.0 mg l^–1 BA, and decreased as the BA concentration increased. IBA was most effective in promoting root formation. The highest rooting frequency (73%) of microshoots was recorded for explants 4.0 cm in length, 4 weeks after culturing. In vitro-rooted plantlets were transferred to polyethylene pots filled with a 1:1:1 (v/v/v) mixture of soil, sand and peat.This treatment resulted in 90% survival of plantlets, which were acclimatised in a greenhouse.