This study aimed to introduce a stress-related gene to improve tolerance of one Egyptian wheat cultivar (Giza164) to oxidative stress, i.e., drought and salinity. This goal was achieved by introducing plant expression vector pLe-GST /bar harboring one of oxidative stress tolerance-related genes Le-GST into the genome using immature embryo-derived calli as a target tissue and microprojectile bombardment as a transformation device. However, the Le-GST gene was driving by CaMV 35S promoter and contained bar gene as a selectable marker. The plasmid pLe-GST/bar was constructed by ligating bar gene cassette to the pAM35S/Le-GST-T3. Putative transgenic plants were tested for bar gene expression. Two transgenic plants were shown to resist the herbicide with no necrosis symptoms then chosen for molecular analysis. Presence of both Le-GST and bar genes was confirmed using PCR and PCR/Southern. On expression level, RT-PCR for Le-GST gene showed the expected band with approximate size confirming its expression in three T1 plants. Finally, these transgenic events will be evaluated under drought and/or salt stress using a higher number of individuals and replicates in subsequent generations |
