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Glutathione reductase activity and isoforms in leaves and roots of wheat plants subjected to cadmium stress

机译:镉胁迫下小麦植株叶片和根中谷胱甘肽还原酶活性及同工型

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摘要

The behavior of glutathione reductase (GR, EC 1.6.4.2) activity and isoforms were analyzed in wheat (Triticum aestivum L.) leaves and roots exposed to a chronic treatment with a toxic cadmium (Cd) concentration. A significant growth inhibition (up to 55%) was found in leaves at 7, 14 and 21 days, whereas roots were affected (51%) only after three weeks. Wheat plants grown in the presence of 100muM Cd showed a time-dependent accumulation of this metal, with Cd concentration being 10-fold higher in roots than in leaves. Nevertheless, lipid peroxidation was augmented in leaves in all experiments, but not in roots until 21 days. Cadmium treatment altered neither the GR activity nor the isoform pattern in the leaves. However, GR activity increased 111% and 200% in roots at 7 and 14 days, respectively, returning to control levels after 21 days. Three GR isoforms were found in roots of control and treated plants, two of which were enhanced by Cd treatment at 7 and 14 days, as assessed by activity staining on native gels. The changes in the isoform pattern modified the global kinetic properties of GR, thereby decreasing significantly (2.5-fold) the Michaelis constant (K(m)) value for oxidized glutathione. Isozyme induction was not associated with an enhancement of GR mRNA and protein expression, indicating that post-translational modification could occur. Our data demonstrated that up-regulation of GR activity by the induction of distinctive isoforms occurs as a defense mechanism against Cd-generated oxidative stress in roots.
机译:分析了暴露于有毒镉(Cd)浓度长期处理的小麦(Triticum aestivum L.)叶片和根中谷胱甘肽还原酶(GR,EC 1.6.4.2)活性和同工型的行为。在第7、14和21天,叶片中发现了显着的生长抑制作用(高达55%),而仅在三周后根受到了影响(51%)。在100μMCd存在下生长的小麦植株显示出这种金属的时间依赖性积累,根中Cd浓度比叶中高10倍。然而,在所有实验中,脂质过氧化作用均在叶片中增加,但直到21天才在根部增加。镉处理既不改变叶片的GR活性,也不改变其同工型。但是,在第7天和第14天,根部的GR活性分别增加111%和200%,在21天后恢复到对照水平。在对照和处理过的植物的根中发现了三种GR同工型,其中7种和14天通过Cd处理增强了两种同工型,这是通过在天然凝胶上进行活性染色来评估的。同工型模式的变化修改了GR的整体动力学特性,从而显着降低了氧化谷胱甘肽的米氏常数(K(m))值(2.5倍)。同工酶诱导与GR mRNA和蛋白表达的增强无关,表明可能发生翻译后修饰。我们的数据表明,通过诱导独特的同工型而引起的GR活性上调是针对根部Cd产生的氧化应激的防御机制。

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