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铝诱导洋葱根细胞氧化爆发、细胞壁 NADH 过氧化物酶活性和 DNA 损伤。

Aluminum induces oxidative burst, cell wall NADH peroxidase activity, and DNA damage in root cells of Allium cepa L.

机构信息

Department of Botany, Molecular Biology and Genomics Laboratory, Berhampur University, Berhampur, India.

出版信息

Environ Mol Mutagen. 2012 Aug;53(7):550-60. doi: 10.1002/em.21719. Epub 2012 Aug 2.

Abstract

Plants under stress incur an oxidative burst that involves a rapid and transient overproduction of reactive oxygen species (ROS: O(2) (•-) , H(2) O(2) , (•) OH). We hypothesized that aluminum (Al), an established soil pollutant that causes plant stress, would induce an oxidative burst through the activation of cell wall-NADH peroxidase (NADH-PX) and/or plasma membrane-associated NADPH oxidase (NADPH-OX), leading to DNA damage in the root cells of Allium cepa L. Growing roots of A. cepa were treated with Al(3+) (800 μM of AlCl(3) ) for 3 or 6 hr without or with the pretreatment of inhibitors specific to NADH-PX and NADPH-OX for 2 hr. At the end of the treatment, the extent of ROS generation, cell death, and DNA damage were determined. The cell wall-bound protein (CWP) fractions extracted from the untreated control and the Al-treated roots under the aforementioned experimental conditions were also subjected to in vitro studies, which measured the extent of activation of peroxidase/oxidase, generation of (•) OH, and DNA damage. Overall, the present study demonstrates that the cell wall-bound NADH-PX contributes to the Al-induced oxidative burst through the generation of ROS that lead to cell death and DNA damage in the root cells of A. cepa. Furthermore, the in vitro studies revealed that the CWP fraction by itself caused DNA damage in the presence of NADH, supporting a role for NADH-PX in the stress response. Altogether, this study underscores the crucial function of the cell wall-bound NADH-PX in the oxidative burst-mediated cell death and DNA damage in plants under Al stress.

摘要

在胁迫下,植物会发生氧化爆发,涉及活性氧物种(ROS:O2(•-)、H2O2、(•)OH)的快速和短暂的过度产生。我们假设,铝(Al)是一种已被确定的土壤污染物,会导致植物胁迫,它会通过激活细胞壁-NADH 过氧化物酶(NADH-PX)和/或质膜相关的 NADPH 氧化酶(NADPH-OX)来诱导氧化爆发,导致洋葱根细胞的 DNA 损伤。洋葱根在没有或用 NADH-PX 和 NADPH-OX 的抑制剂预处理 2 小时的情况下,用 800 μM 的 AlCl3 处理 3 或 6 小时。在处理结束时,测定 ROS 的产生、细胞死亡和 DNA 损伤的程度。还对未经处理的对照和上述实验条件下用 Al 处理的根的细胞壁结合蛋白(CWP)部分进行了体外研究,这些研究测量了过氧化物酶/氧化酶的激活程度、(•)OH 的产生和 DNA 损伤的程度。总的来说,本研究表明,细胞壁结合的 NADH-PX 通过产生导致洋葱根细胞死亡和 DNA 损伤的 ROS,有助于 Al 诱导的氧化爆发。此外,体外研究表明,在存在 NADH 的情况下,CWP 部分本身会导致 DNA 损伤,支持 NADH-PX 在应激反应中的作用。总之,这项研究强调了细胞壁结合的 NADH-PX 在 Al 胁迫下植物中氧化爆发介导的细胞死亡和 DNA 损伤中的关键作用。

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