Efficient transformation of wheat by using a mutated rice acetolactate synthase gene as a selectable marker
Efficient transformation of wheat by using a mutated rice acetolactate synthase gene as a selectable marker
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DOI：
10.1007/s00299-008-0553-6
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发表时间：
2008-08-01
影响因子：
6.2
通讯作者：
Handa, Hirokazu
中科院分区：
文献类型：
--
作者：
Ogawa, Taiichi;Kawahigashi, Hiroyuki;Handa, Hirokazu
乙酰乳酸合成酶（ALS）是许多除草剂的靶标酶，包括磺酰脲和咪唑啉酮。我们研究了突变的水稻ALS基因的有用性，该基因具有双点突变并编码抗除草剂形式的酶，作为小麦转化的选择标记。利用基因枪法将含有ALS基因突变体的水稻基因组DNA片段导入幼胚，用除草剂双草醚钠（BS）筛选出转基因植株。Southern印迹分析证实转基因植株的染色体上有1至10个以上的转基因拷贝。BS浓度的调整结合重复选择有效地防止非转基因植物逃避除草剂的选择。ALS活性测定表明，转基因植株产生了抗除草剂的ALS，因此获得了对BS的抗性。这是第一份描述利用植物来源的选择标记基因进行小麦转化的选择系统的报告。
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.