Wakasa Yuhya, Ozawa Kenjirou, Takaiwa Fumio
Transgenic Crop Research and Development Center, National Institute of Agrobiological Sciences, Kannondai 3-1-3, Tsukuba, Ibaraki 305-8602, Japan.
Plant Cell Rep. 2007 Sep;26(9):1567-73. doi: 10.1007/s00299-007-0373-0. Epub 2007 May 22.
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.
我们开发了一种仅使用水稻基因组衍生成分的高效水稻转化系统。转基因“越光”水稻低谷蛋白突变体a123能够在胚乳中积累大量生物活性肽。以愈伤组织特异性启动子(CSP)控制下表达的突变乙酰乳酸合酶(mALS)基因作为选择标记,通过农杆菌介导的转化将绿色荧光蛋白(GFP)和一种抗高血压六肽导入“越光”a123。CSP:mALS基因盒赋予转基因水稻愈伤组织嘧啶基羧基除草剂抗性,但在再生植株中不表达。通过改良传统用于水稻组织培养的愈伤组织诱导、愈伤组织筛选和再生培养基,转基因水稻品系a123的转化效率从约10%提高到了约30%。