This research was conducted on Stevia rebaudiana which propagated in vitro to study the effect of pH on propagation, plant growth, rooting and to determine the optimum pH of the media. The research aimed to determine the concentration of copper sulphate in the media after determining the best pH. MS media (Murashige and Skoog, 1962) was used in all stages of the experiment and different pH (4, 4.5, 5, 5.5, and 5.8), in the second experiment used different concentrations of copper sulphate (0.10 µM, 0.15 µM, 0.20 µM and 0.25 µM), all the media were without hormones, the best media for the propagation, growth and rooting stevia rebaudiana was MS pH 4.5 Statistical analysis showed statistically significant superiority to the recipe the long of branches in MS pH4.5 the average of length was 8 cm also outperformed in the average number of branches and average length of roots. The study also demonstrated that the best concentration of copper sulphate in the media MSpH4.5 was 0.15 µM and statistical analysis showed that this concentration in the recipe the long and the number of branches and the number of roots at all concentrations has been shown that plant growth in MSpH4.5 and 0.15 µM concentration of copper sulphate was the best in all studied media that we get. The whole plant is ready for transfer and hardening in the greenhouse after 30 days of in vitro cultivation. All readings were recorded for both experiments after four weeks of in vitro cultivation in test tubes.