Citrus is one of the most important commodity worldwide, owing to its tremendous nutritional value, and acceptable as fresh edible food. This study was conducted at the Desert Research Center (DRC) in Cairo, Egypt, during the period 2013-2017. An efficient in vitro propagation system for Volkamer lemon (Citrus volkameriana) was established. The effect of various combination of two plant growth regulators (cytokinin and auxin) was evaluated on the proliferation efficiency of citrus plant via in vitro propagation technique. The sterilized stem nodal segments of the given species were planted vertically on MS culture medium augmented with various combinations of BAP at 0.0, 0.5, 1.0, 1.5, 2.0, 2.5, and 3.0 mg/l and NAA at 0.0, 0.1, and 0.2 mg/l, and especially at BAP×NAA at 2.0 ×0.1 mg/l respectively proved to be the best for bud induction. The survival percentage of stem node segment frequency was 100%, especially when MS medium was supplemented with BAP in a range 1.0- 3.0 mg/l. The best shoot initiation was obtained on MS medium augmented with BAP and NAA at 2.0 and 0.1 mg/l, each in turn, which recorded the highest mean value (3.99 shoots/ explant). Higher number of elongated shoots was obtained on MS medium with BAP and NAA at 3.0 and 0.1 mg/l, in order. The higher multiplication rate was recorded on MS medium containing BAP and NAA at either 2.0 or 2.5 mg/l for the former and 0.05 mg/l for the latter. The shoots were then rooted on MS medium containing IBA and NAA at 1.5 and 0.2 mg/l consecutively, with high rooting percentage (100%). The plantlets survival ex vitro swas (80%) when plantlets were transferred to plastic pots containing a mixture of sand and peatmoss (1:1). In conclusion, this study provides reproducible technique for micropropagation of Volkamer Lemon.