The present study was carried out to investigate the influence of different types, combinations and concentrations of plant growth regulators added to Murashige and Skoog (MS) medium on establishment, shoot multiplication and rooting of pistachio (Pistacia vera L.) rootstock. An efficient and rapid method for micropropagation of P. vera was developed using stem node sections which were collected in the spring from adult trees grown in El-Maghara Experimental Station (Middle Sinai). The explants were cultured on MS medium supplemented with different concentrations of 6- benzyl adenin (BA) (0.0,0.5,1.0, and 1.5 mg/l) in combination with β-naphthalene acetic acid (NAA)(0.0, 0.01 and 0.05 mg /l). Obtained shoots from establishment stage were placed on medium supplemented with various types of cytokinins BA, kinetin (Kin) and Thiodiazuron (TDZ) each at different concentrations (0.5, 1.0, 1.5 and 2.0 mg/ l). The highest average number of shoots and shoot length (4.7 and 4.2 cm, respectively) were obtained when shoots were placed on MS medium supplemented with 1 mg/l BA. However, MS medium supplemented with TDZ at 0.5 mg /l raised the average shoot number to 3.8 and average length to 3.9 cm. Root initiation and growth was obtained on half strength MS medium supplemented with 2.0 mg/l, indol-3-butyric acid (IBA). IBA proved to be more effective for rooting than NAA. In vitro formed plantlets were left for a month in jars under lab conditions till forming a whole rooting system. Rooted pistachio plantlets were acclimatized successfully following the regular methods. Such a protocol will be helpful to expand the cultivation of pistachio in Sinai.