A novel mutated acetolactate synthase gene conferring specific resistance to pyrimidinyl carboxy herbicides in rice
A novel mutated acetolactate synthase gene conferring specific resistance to pyrimidinyl carboxy herbicides in rice
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DOI：
10.1007/s11103-007-9146-y
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发表时间：
2007-05-01
影响因子：
5.1
通讯作者：
Toriyama, Kinya
中科院分区：
文献类型：
--
作者：
Okuzaki, Ayako;Shimizu, Tsutomu;Toriyama, Kinya
乙酰乳酸合成酶（ALS）是支链氨基酸生物合成途径中最常见的酶。已知ALS中特定氨基酸的突变会赋予对ALS抑制除草剂（例如磺酰脲类和嘧啶羧基（PC）除草剂）的抗性。然而，迄今为止尚未报道赋予对PC的排他性抗性的突变。以水稻花药培养的愈伤组织为材料，用双草醚钠（BS）作为筛选剂，筛选出抗PC的水稻愈伤组织。获得了两株携带ALS中第95位甘氨酸突变为丙氨酸（G95 A）的新突变的BS抗性植物。体外ALS活性测定表明，G95 A突变的ALS重组蛋白（ALS-G95 A）对PC除草剂具有高度的特异性抗性。为了确定ALS-G95 A基因是否可用作水稻转化的选择标记，将ALS-G95 A基因连接到泛素启动子并导入水稻。以BS为筛选剂，筛选到了整合有ALS-G95 A基因的PC抗性植株。总之，新的G95 A突变的ALS基因赋予对PC-除草剂的高度特异性抗性，并且可以用作选择标记。
Acetolactate synthase (ALS) is the first common enzyme in the biosynthetic pathway of branched-chain amino acids. Mutations of specific amino acids in ALS have been known to confer resistance to ALS-inhibiting herbicides such as sulfonylureas and pyrimidinyl carboxy (PC) herbicides. However, mutations conferring exclusive resistance to PC have not yet been reported to date. We selected PC resistant rice calli, which were derived from anther culture, using one of the PCs, bispyribac-sodium (BS), as a selection agent. Two lines of BS-resistant plants carrying a novel mutation, the 95th Glycine to Alanine (G95A), in ALS were obtained. In vitro ALS activity assay indicated that the recombinant protein of G95A-mutated ALS (ALS-G95A) conferred highly specific resistance to PC herbicides. In order to determine if the ALS-G95A gene could be used as a selection marker for rice transformation, the ALS-G95A gene was connected to ubiquitin promoter and introduced into rice. PC resistant plants containing integrated ALS-G95A gene were obtained after selection with BS as a selection agent. In conclusion, novel G95A mutated ALS gene confers highly specific resistant to PC-herbicides and can be used as a selection marker.