Agrobacterium-mediated transformation of a low glutelin mutant of 'Koshihikari' rice variety using the mutated-acetolactate synthase gene derived from rice genome as a selectable marker

We have developed an efficient rice transformation system that uses only rice genome-derived components. The transgenic 'Koshihikari' rice, low-glutelin mutant a123, is capable of accumulating large amounts of bioactive peptides in the endosperm. Agrobacterium-mediated transformation using the mutated-acetolactate synthase (mALS) gene expressed under the control of the callus-specific promoter (CSP) as a selectable marker was used to introduce GFP and an anti-hypertensive hexapeptide into 'Koshihikari' a123. The CSP:mALS gene cassette confers pyrimidinyl carboxy herbicide resistance to transgenic rice callus, but is not expressed in regenerated plants. Transformation efficiency of transgenic rice line a123 was improved from about 10% to about 30% by modifying callus induction, callus selection and regeneration media conventionally used for rice tissue culture.