The present study was conducted at the tissue culture lab. Center of Genetic Engineering and Biotechnology, Faculty of Agriculture, Ain Shams University, Egypt during the period from 2020 to 2022 to study the effect of plant growth regulators and their concentration on the multiplication stage of a strawberry (Fragaria x ananassa Duch) cultivars, i.e. Fortuna, through tissue culture technique. Data showed that Fortuna cv. The medium amended with kinetin at concentrations of (5 mg/l) was the best for giving the highest number of shoots/explant, while the highest length of shoots/explant was observed when BA at concentrations of (0.5 mg/l) and IBA at concentrations of (1 mg/l) during the multiplication stage.