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The ipt-type MAT vector uses the ipt gene for regeneration of marker-free transgenic plants. However, it was pointed out that this system was not suitable for most economically important crops that regenerated through auxin-dependent embryogenesis. We report a single-step transformation system of rice using MAT vector. When we transformed scutellum tissues of 5 days pre-cultured rice seeds, marker-free transgenic rice plants directly regenerated from 25.5% infected scutellum tissues without forming ipt-intermediates within 4 weeks after an infection. Excision of the ipt gene caused the regeneration of marker-free transgenic rice plants through embryogenic tissues. Therefore, this system needs no selective agent and no sexual crossing for identification of transgenic plants not containing a selectable marker gene. This system is highly effective for generation of marker-free transgenic plants in economically important crops.