This study is conducted to find out the effect of genetic transformation by Agrobacterium rhizogenes ATCC13332 on the anatomical features of Eruca sativa. Stem explants and leaves were inoculated by direct injection of A. rhizogenes. The results indicated the high response of E. sativa, as the percentage of hairy roots formation was 95.3% for leaves. Callus was successfully initiated from hairy roots on both Murashige-Skoog (MS) and Woody plants (WP) media containing (0.4 and 0.5 mg. L-1 thidiazuron(TDZ). Shoot regeneration occurred spontaneously. Also, the callus originated from hairy roots showed ability of shooting. Genetic transformation was detected by polymerase chain reaction (PCR). Alterations in anatomical structure of stems and leaves in transformed plants were observed. Transgenic plants' epidermal cell thickness was 14.3 µm rather than 12 µm. There are nine vascular bundles in regenerated plants that are open collateral vascular bundles. The palisade tissue thickness of transformed leaves was 96 µm, while 66 µm in non-transformed. It is clear that the genetic transformation led to anatomical changes of this medicinal plant.