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Efficient transformation of wheat by using a mutated rice acetolactate synthase gene as a selectable marker

机译:以突变的水稻乙酰乳酸合酶基因为选择标记对小麦进行高效转化

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摘要

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.
机译:乙酰乳酸合酶(ALS)是许多除草剂(包括磺酰脲和咪唑啉酮)的目标酶。我们调查了水稻突变的ALS基因的有用性,该基因具有双点突变并编码了除草剂抗性形式的酶,作为小麦转化的选择标记。在通过粒子轰击将来自水稻的含有突变ALS基因的基因组DNA片段引入未成熟胚后,用除草剂双嘧菌钠(BS)有效地选择了转基因植物。 Southern印迹分析证实,转基因植物的染色体中具有一至十个以上的转基因拷贝。调节BS浓度并重复选择有效地防止了非转基因植物逃脱除草剂选择。 ALS活性的测量表明转基因植物产生了抗除草剂形式的ALS,因此获得了对BS的抗性。该报告是第一个描述使用植物来源选择标记基因的小麦转化选择系统的报告。

著录项

  • 来源
    《Plant Cell Reports》 |2008年第8期|1325-1331|共7页
  • 作者单位

    Plant Genome Research Unit National Institute of Agrobiological Sciences Kan-non-dai 2-1-2 Tsukuba Ibaraki 305-8602 Japan;

    Plant Genome Research Unit National Institute of Agrobiological Sciences Kan-non-dai 2-1-2 Tsukuba Ibaraki 305-8602 Japan;

    Plant Genetic Engineering Research Unit National Institute of Agrobiological Sciences Kan-non-dai 2-1-2 Tsukuba Ibaraki 305-8602 Japan;

    Plant Genome Research Unit National Institute of Agrobiological Sciences Kan-non-dai 2-1-2 Tsukuba Ibaraki 305-8602 Japan;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    Herbicide tolerance; Particle bombardment; Selection; Transgenic plant; Triticum aestivum;

    机译:除草剂耐受性;颗粒轰击;选择;转基因植物;普通小麦;

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