This current study was conducted on gardenia plant (Gardenia jasminoides.L.) cv. ellis. which was micropropagated in vitro to study the effect of some plant growth regulators on multiplication and rooting, for the aim of developing a complete in vitro microprpagation system of the studied cv. Results demonstrated that, the best medium for in vitro micropropagation of the studied cv. was MS medium supplemented with 1 mg/L BA + 0.1 mg/L IBA with a multiplication rate of 5.33 new shoots, with average shoot height of 4.73 cm, and average leaf number of 4.36. These shoots were then transferred to a rooting media containing a half-strength of the multiplication basal media with NAA at a concentration of 1mg/L which resulted in highest rooting efficiency of 97% with average root number of 9.67, and average root length of 5.35 cm, and average stem length of 9.80 cm, and average leaf number of 5.07. Increasing NAA concentration in rooting media to 1.5 mg/L resulted in increasing root length average to 6.28 cm with significant difference in comparison to the previous treatment. Rooted Plantlets were acclimatized gradually in a growth room conditions to ex vitro conditions with 85 % efficiency.