Investigation of transformation and expression of roseltide rT1 gene in wheat

Antimicrobial peptides display powerful antimicrobial activities against a large set of plant pathogens and are a good alternative to conventional toxins. To transfer rT1 gene of Hibiscus sabdariffa to wheat, Agrobacterium tumefaciens strain LBA4404 harboring the pBI121-rT1 plasmid was used. Mature embryo explants of four wheat cultivars including the Saison, Bahar, Chamran and Arta after co-cultivated with A. tumefaciens suspension were transferred to a selective callus medium containing 2 mg/l 2,4-D, 2 mg/l Picloram, 50 mg/l Kanamycin and 500 mg/l Cefotaxime. Embryogenic calli were selected after growth in a selective medium and then transferred to a regenerative medium containing 25 mg/l Kanamycin and 250 mg/l Cefotaxime. The presence and expression of rT1 gene at the transcriptional level in transgenic plants were confirmed by PCR and RT-PCR methods respectively using specific primers. Based on the RT-PCR results, the rT1 gene was expressed in transgenic wheat plants. In this study, out of 15 possible transgenic plants, only 11 were transgenic. Among transgenic plants, five plants belonged to Saison cultivar with 1.6 transgenic percentage, four plants represented to the Bahar cultivar with 1.3 transgenic percentage and two plants belonged to the Chamran cultivar with 0.6 transgenic percentage. No transgenic plant was obtained for the Arta cultivar.