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A method of Agrobacterium tumefaciens mediated transformation for perennial ryegrass was developed using the calli of ryegrass derived from mature embryos. The calli were inoculated with a disarmed A. tumefaciens strain EHA105 harboring binary vector p2328. Vector p2328 contained transcription factor DREB1B and neomycin phosphotransferase (npt II) genes which were driven by promoters of rd29B and CaMV35S, respectively. The inoculated calli were selected on paromomycin- or kanamycin-containing media till the established plants being transferred to soil. Six transgenic plants with DREB1B had been obtained from perennial ryegrass strain Tove. PCR and Southern-blotting showed that npt II and DREB1B genes were integrated in perennial ryegrass genome. Stress treatment confirmed that transgenic plants with higher drought tolerance were obtained.