Transformation of Arabidopsis by mutated acetolactate synthase genes from rice and Arabidopsis that confer specific resistance to pyrimidinylcarboxylate-type ALS inhibitors
Transformation of Arabidopsis by mutated acetolactate synthase genes from rice and Arabidopsis that confer specific resistance to pyrimidinylcarboxylate-type ALS inhibitors
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DOI：
10.5511/plantbiotechnology.27.75
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发表时间：
2010-01-01
影响因子：
1.6
通讯作者：
Shimizu, Tsutomu
中科院分区：
文献类型：
--
作者：
Kawai, Kiyoshi;Kaku, Koichiro;Shimizu, Tsutomu
之前，我们发现了来自水稻和拟南芥的四个突变的乙酰乳酸合成酶（ALS）基因（W548L/S627IOsALS, S627IOsALS， W574L/S653IAtALS和S653IAtALS）赋予了对嘧啶羧酸盐型ALS抑制剂的高水平抗性（Kawai et al. 2008）。从存在ALS抑制剂的水稻细胞中获得了突变的ALS基因。人工合成的拟南芥ALS突变基因具有与水稻相同的氨基酸替换。在这里，我们证明了这些突变基因是拟南芥转化的有效选择标记。具体来说，我们研究了突变的ALS在拟南芥中的表达及其对转基因拟南芥植株对ALS抑制剂敏感性的影响。我们的研究结果表明，表达拟南芥突变ALS的转化子对ALS抑制剂的抗性程度高于表达水稻突变ALS的转化子。在系统发育树上，单子叶植物和双子叶植物的氨基酸序列被划分为两个簇。综上所述，利用水稻突变的ALS基因和拟南芥突变的ALS基因分别培育单子叶和双子叶转基因植株较为合适。此外，我们的发现在产生具有已知ALS核苷酸序列的转基因植物时特别有用。在这种情况下，携带这些突变的ALS基因可以作为选择性标记，因为突变点的氨基酸残基在植物物种中是保守的。
Previously, we showed that four mutated acetolactate synthase (ALS) genes derived from rice and Arabidopsis (W548L/S627IOsALS, S627IOsALS, W574L/S653IAtALS and S653IAtALS) confer high levels of resistance to pyrimidinylcarboxylate type ALS inhibitors (Kawai et al. 2008). Mutated ALS genes of rice were obtained from rice cells cultured in the presence of an ALS-inhibitor. The mutated ALS genes of Arabidopsis, which have the same amino acid substitutions as those of rice, have been generated artificially. Here, we demonstrate that these mutated genes function as effective selectable markers for transformation of Arabidopsis. Specifically, we studied expression of the mutated ALSs in Arabidopsis and their effect on the sensitivity of transgenic Arabidopsis plants to the ALS inhibitors. Our results show that the degree of resistance to the ALS inhibitors of transformants expressing Arabidopsis mutated ALSs was greater than those of transformants expressing rice mutated ALSs. The amino acid sequences of ALSs derived from monocotyledonous plants and those derived from dicotyledonous plants were clearly divided into two clusters in a phylogenetic tree. Based on these results, it would be preferable to use rice and Arabidopsis mutated ALS genes for generating monocotyledonous and dicotyledonous transgenic plants, respectively. Moreover, our findings are particularly useful when generating transgenic plants with a known ALS nucleotide sequence. In such cases, their own ALS gene carrying these mutations could be used as a selectable marker because amino acid residues at the point of mutation are conserved among plant species.