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利用转基因水稻植株分析猪CYP2B22和CYP2C49对除草剂的底物特异性。

Analysis of substrate specificity of pig CYP2B22 and CYP2C49 towards herbicides by transgenic rice plants.

作者信息

Kawahigashi Hiroyuki, Hirose Sakiko, Ozawa Kenjirou, Ido Yoshiko, Kojima Misaki, Ohkawa Hideo, Ohkawa Yasunobu

机构信息

Plant Biotechnology Department, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, Japan.

出版信息

Transgenic Res. 2005 Dec;14(6):907-17. doi: 10.1007/s11248-005-0199-x.

Abstract

We introduced two novel types of pig (Sus scrofa) cytochrome P450, CYP2B22 and CYP2C49, into rice plants (Oryza sativa L. cv. 'Nipponbare') to produce herbicide-tolerant plants and to confirm the metabolic activities of the cytochrome P450 species. In germination tests, both types of transgenic plants showed tolerance to various herbicides with different modes of action. CYP2B22 rice plants showed tolerance towards 12 herbicides including chlortoluron (100 microM), amiprofos-methyl (2.5 microM), pendimethalin (10 microM), metolachlor (2.5 microM), and esprocarb (20 microM). CYP2C49 rice plants showed tolerance towards 13 herbicides, including chlortoluron (100 microM), norflurazon (0.5 microM), amiprofos-methyl (2.5 microM), alachlor (0.8 microM), and isoxaben (1 microM). The herbicide tolerance was considered to reflect the substrate specificity of the introduced P450 species. We used (14)C-labeled metolachlor and norflurazon to confirm the P450 activity in the transgenic rice plants. The herbicides were metabolized more quickly in the transgenic rice plants than in the nontransgenic rice plants. Therefore, CYP2B22 and CYP2C49 rice plants became more tolerant to various herbicides than nontransgenic control plants because of accelerated metabolism of the herbicides by the introduced P450 species. Assuming that public and commercial acceptance is forthcoming, these transgenic rice plants may become useful tools for the breeding of herbicide-tolerant crops.

摘要

我们将两种新型猪(野猪)细胞色素P450,即CYP2B22和CYP2C49,导入水稻植株(水稻品种‘日本晴’)中,以培育耐除草剂植物并确认细胞色素P450物种的代谢活性。在发芽试验中,两种转基因植株对具有不同作用模式的多种除草剂均表现出耐受性。表达CYP2B22的水稻植株对12种除草剂具有耐受性,包括绿麦隆(100微摩尔)、丙草胺(2.5微摩尔)、二甲戊灵(10微摩尔)、异丙甲草胺(2.5微摩尔)和丙草丹(20微摩尔)。表达CYP2C49的水稻植株对13种除草剂具有耐受性,包括绿麦隆(100微摩尔)、氟乐灵(0.5微摩尔)、丙草胺(2.5微摩尔)、甲草胺(0.8微摩尔)和异恶草松(1微摩尔)。除草剂耐受性被认为反映了导入的P450物种的底物特异性。我们使用(14)C标记的异丙甲草胺和氟乐灵来确认转基因水稻植株中的P450活性。与非转基因水稻植株相比,除草剂在转基因水稻植株中的代谢更快。因此,由于导入的P450物种加速了除草剂的代谢,表达CYP2B22和CYP2C49的水稻植株比非转基因对照植株对各种除草剂的耐受性更强。假设能够获得公众和商业认可,这些转基因水稻植株可能成为培育耐除草剂作物的有用工具。

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