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来自詹氏菌属的一种新型天然I类5-烯醇丙酮酸莽草酸-3-磷酸合酶赋予水稻高草甘膦耐受性。

A Novel Naturally Occurring Class I 5-Enolpyruvylshikimate-3-Phosphate Synthase from Janibacter sp. Confers High Glyphosate Tolerance to Rice.

作者信息

Yi Shu-yuan, Cui Ying, Zhao Yan, Liu Zi-duo, Lin Yong-jun, Zhou Fei

机构信息

State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.

National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research, Huazhong Agricultural University, Wuhan 430070, China.

出版信息

Sci Rep. 2016 Jan 12;6:19104. doi: 10.1038/srep19104.

Abstract

As glyphosate is a broad spectrum herbicide extensively used in agriculture worldwide, identification of new aroA genes with high level of glyphosate tolerance is essential for the development and breeding of transgenic glyphosate-tolerant crops. In this study, an aroA gene was cloned from a Janibacter sp. strain isolated from marine sediment (designated as aroAJ. sp). The purified aroAJ. sp enzyme has a Km value of 30 μM for PEP and 83 μM for S3P, and a significantly higher Ki value for glyphosate (373 μM) than aroAE. coli. AroAJ. sp is characterized as a novel and naturally occurring class I aroA enzyme with glyphosate tolerance. Furthermore, we show that aroAJ. sp can be used as an effective selectable marker in both japonica and indica rice cultivar. Transgenic rice lines were tested by herbicide bioassay and it was confirmed that they could tolerate up to 3360 g/ha glyphosate, a dosage four-fold that of the recommended agricultural application level. To our knowledge, it is the first report of a naturally occurring novel class I aroA gene which can be efficiently utilized to study and develop transgenic glyphosate-tolerant crops, and can facilitate a more economical and simplified weed control system.

摘要

由于草甘膦是一种在全球农业中广泛使用的广谱除草剂,鉴定具有高草甘膦耐受性的新aroA基因对于转基因抗草甘膦作物的开发和育种至关重要。在本研究中,从一株从海洋沉积物中分离的詹氏菌属菌株(命名为aroAJ.sp)中克隆了一个aroA基因。纯化后的aroAJ.sp酶对磷酸烯醇式丙酮酸(PEP)的Km值为30μM,对3-磷酸莽草酸(S3P)的Km值为83μM,并且对草甘膦的Ki值(373μM)明显高于aroAE.coli。aroAJ.sp被鉴定为一种具有草甘膦耐受性的新型天然I类aroA酶。此外,我们表明aroAJ.sp可作为粳稻和籼稻品种的有效选择标记。通过除草剂生物测定对转基因水稻品系进行了测试,证实它们能够耐受高达3360g/ha的草甘膦,这一剂量是推荐农业应用水平的四倍。据我们所知,这是首次报道一种天然存在的新型I类aroA基因,它可有效用于研究和开发转基因抗草甘膦作物,并有助于建立更经济、更简化的杂草控制系统。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7846/4709569/f4467c9a305f/srep19104-f1.jpg

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