Agrobacterium-Mediated Transformation of Pomegranate (Punica granatum L.) ‘Yousef Khani’ Using the gus Reporter Gene

In this study, an efficient agrobacterium-mediated transformation method was developed for pomegranate (punica granatum l.), a difficult-to-transform plant. in vitro shoot segments were inoculated with agrobacterium tumefaciens strain lba4404 harboring the binary vector pbi121 carrying the neomycin phosphotransferase (nptii) gene as a selectable marker and b-glucuronidase (gus) gene as a reporter. after 28 d in wpm selection medium containing 50 mg l-1 kanamycin, 59 new shoots proliferated. gus analysis was performed on these putative transgenic shoots, of which 32 stained positive. positive staining shoots were cut and cultured in selection medium for 2 subsequent subcultures until final gus analysis. after three months of the selection period, 6 putative transgenic shoots were obtained. presence of the gus and nptii genes was confirmed by polymerase chain reaction. southern blot analysis confirmed that t-dna was stably integrated into the genome of three out of six pcr-positive plants. the transgenic plants were rooted and successfully acclimatized.