利用突变的水稻乙酰乳酸合成酶基因作为选择标记对小麦进行高效转化。
Efficient transformation of wheat by using a mutated rice acetolactate synthase gene as a selectable marker.
作者信息
Ogawa Taiichi, Kawahigashi Hiroyuki, Toki Seiichi, Handa Hirokazu
机构信息
Plant Genome Research Unit, National Institute of Agrobiological Sciences, Kan-non-dai 2-1-2, Tsukuba, Ibaraki, 305-8602, Japan.
出版信息
Plant Cell Rep. 2008 Aug;27(8):1325-31. doi: 10.1007/s00299-008-0553-6. Epub 2008 May 1.
Acetolactate synthase (ALS) is a target enzyme for many herbicides, including sulfonylurea and imidazolinone. We investigated the usefulness of a mutated ALS gene of rice, which had double point mutations and encoded an herbicide-resistant form of the enzyme, as a selectable marker for wheat transformation. After the genomic DNA fragment from rice containing the mutated ALS gene was introduced into immature embryos by means of particle bombardment, transgenic plants were efficiently selected with the herbicide bispyribac sodium (BS). Southern blot analysis confirmed that transgenic plants had one to more than ten copies of the transgene in their chromosomes. Adjustment of the BS concentration combined with repeated selection effectively prevented nontransgenic plants from escaping herbicide selection. Measurement of ALS activity indicated that transgenic plants produced an herbicide-resistant form of ALS and therefore had acquired the resistance to BS. This report is the first to describe a selection system for wheat transformation that uses a selectable marker gene of plant origin.
乙酰乳酸合酶(ALS)是包括磺酰脲类和咪唑啉酮类在内的许多除草剂的作用靶标酶。我们研究了水稻中一个具有双点突变且编码该酶抗除草剂形式的突变ALS基因作为小麦转化选择标记的实用性。通过微粒轰击将含有突变ALS基因的水稻基因组DNA片段导入未成熟胚后,用除草剂双草醚(BS)高效筛选出了转基因植株。Southern杂交分析证实转基因植株的染色体中有1到10多个转基因拷贝。调整BS浓度并结合反复筛选有效地防止了非转基因植株逃避除草剂筛选。ALS活性测定表明转基因植株产生了抗除草剂形式的ALS,因此获得了对BS的抗性。本报告首次描述了一种利用植物源选择标记基因的小麦转化选择系统。