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Organ-Specific Expression of Glutathione S-Transferases and the Efficacy of Herbicide Safeners in Arabidopsis

机译:拟南芥中谷胱甘肽S-转移酶的器官特异性表达和除草剂安全剂的功效

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摘要

The functions of plant glutathione S-transferases (GSTs) under normal growth conditions are poorly understood, but their activity as detoxification enzymes has been harnessed in agriculture for selective weed control. Herbicide safeners protect monocot crops from herbicide injury but have little effect on weedy monocot or dicot species. Protection by safeners is associated with expression of herbicide-metabolizing enzymes including GSTs, but the basis for selective action of safeners between monocots and dicots is not known. To address this question we have studied the response of Arabidopsis (Arabidopsis thaliana) to various safeners. Benoxacor, fenclorim, and fluxofenim did not protect Arabidopsis from herbicide injury but did induce RNA expression of the glutathione-conjugate transporters encoded by AtMRP1, AtMRP2, AtMRP3, and AtMRP4. These safeners also induced the organ-specific expression of AtGSTU19 and AtGSTF2, two previously characterized Arabidopsis GSTs from different classes of this enzyme family. RNA hybridization, immunoblot, and reporter gene analyses indicated expression of AtGSTU19 induced by safeners predominated in roots. To test the hypothesis that increased expression of AtGSTU19 would be sufficient to provide tolerance to chloroacetamide herbicides, a chimeric gene was produced containing the open reading frame for this GST driven by a constitutive promoter. Plants containing this transgene had a modest increase in AtGSTU19 protein, predominantly in roots, but this had no effect on tolerance to chloroacetamide herbicides. The localized induction of GSTs by safeners in roots of Arabidopsis may explain why these compounds are unable to provide herbicide tolerance to dicot plant species.
机译:人们对植物谷胱甘肽S-转移酶(GSTs)在正常生长条件下的功能了解甚少,但在农业中已将其作为排毒酶的活性用于选择性除草。除草剂安全剂可保护单子叶植物免受除草剂的伤害,但对杂草单子叶植物或双子叶植物几乎没有影响。安全剂的保护作用与包括GST在内的除草剂代谢酶的表达有关,但尚不清楚单子叶植物和双子叶植物之间安全剂选择性作用的基础。为了解决这个问题,我们研究了拟南芥(Arabidopsis thaliana)对各种安全剂的反应。 Benoxacor,fenclorim和flufenfenim不能保护拟南芥免受除草剂的伤害,但可以诱导由AtMRP1,AtMRP2,AtMRP3和AtMRP4编码的谷胱甘肽-结合转运蛋白的RNA表达。这些安全剂还诱导了AtGSTU19和AtGSTF2的器官特异性表达,这是该酶家族不同类别的两个先前表征的拟南芥GST。 RNA杂交,免疫印迹和报告基因分析表明,根部占主导地位的安全分子诱导了AtGSTU19的表达。为了测试假设AtGSTU19的表达增加足以提供对氯乙酰胺除草剂的耐受性,产生了一个嵌合基因,该基因包含由组成型启动子驱动的该GST的开放阅读框。含有该转基因的植物的AtGSTU19蛋白适度增加,主要在根部,但这对氯乙酰胺除草剂的耐受性没有影响。安全剂在拟南芥根中对GSTs的局部诱导可能解释了为什么这些化合物无法对双子叶植物物种提供除草剂耐受性。

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