A study to micro-propagate guava (Pisidium guajava) was carried out in 2020 and 2021 at the Tissue Culture Laboratory, Horticulture Department, Faculty of Agriculture, Tanta University. Nodal segments from in vitro germinated seeds of guava were used as explants. Nodal segments (from germinated seeds) were cultured on MS media supplemented with different concentrations of benzyl adenine (BA), kinetin (KIN), or thidiazuron (TDZ) (0, 1 and 2 mg/L). 2 mg/L BA was the most favorable treatment for multiplication of shoots (21.13 shoots/explant) compared to the other treatments and the control (plant growth regulators free medium). Shoots of suitable length were transferred to the rooting media containing (0, 0.5, or 1 mg/L) IBA. The medium containing 0.5 mg/L IBA gave the highest number of roots/plantlets and the longest roots. Under greenhouse for acclimatization, the plantlets of guava derived from the medium supplemented with 0.5 or 1 mg/L indole butyric acid recorded the highest survival percentage (80%). It can be concluded that added 2 mg/L of BA in the medium gave the highest number of shoots/explant during the multiplication stage of guava shoots. In addition, added 0.5 or 1 mg/L IBA during the rooting stage increased the number of roots/plantlets with recorded a highest survival percentage after five weeks of acclimatization stage.