This investigation was conducted on Ru140 grape rootstock at the General Commission for Scientific Agricultural Research (GCSAR), Damascus with the aim of in vitro vegetatively micropropagation using some plant growth regulators on multiplication and rooting to determine the best combinations and concentrations of plant growth regulators that result in the best multiplication rate, and best rooting crekeria (rate and roots number and length). Results demonstrated that, the best medium for in vitro micropropagation of the studied rootstock was the modified MS medium supplemented with 4.44 μM BA + 0.49 μM IBA with multiplication rate of 7.72 new shoots every 4 weeks, and shoots lengt of 5.54 cm. These shoots were transferred for 4 weeks to among elongation medium containing the same medium with the addition of Kinetine at a concentration of 2.22 μM instead of BA which led to a shoot elongation rate of 7.87 cm, then these shoots were transferred to rooting medium for rooting, It was shown that using auxin IBA at a concentration of 4.44 μM resulted at the highest rate of rooting (87%) with the largest number of roots (7.56) when using the auxin IBA concentration μM 4.44 compared with the rest of other transactions and with the control as well. However, The highest length of roots (6.29 cm) was observed on medim contained lower IBA concentration (2.22 μM). Rooted Plants were acclimatized gradually to ex vitro conditions with 70 % efficiency.