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氧化应激通过多平行实时定量 PCR 分析 ROS 标记物和抗氧化基因,在耐受应激的 atr7 和敏感的 loh2 拟南芥突变体中引发明显的转录反应。

Oxidative stress provokes distinct transcriptional responses in the stress-tolerant atr7 and stress-sensitive loh2 Arabidopsis thaliana mutants as revealed by multi-parallel quantitative real-time PCR analysis of ROS marker and antioxidant genes.

机构信息

Department of Plant Physiology and Plant Molecular Biology, University of Plovdiv, 24 Tsar Assen Str., Plovdiv 4000, Bulgaria.

出版信息

Plant Physiol Biochem. 2012 Oct;59:20-9. doi: 10.1016/j.plaphy.2012.05.024. Epub 2012 Jun 4.

DOI:10.1016/j.plaphy.2012.05.024
PMID:22710144
Abstract

The Arabidopsis thaliana atr7 mutant is tolerant to oxidative stress induced by paraquat (PQ) or the catalase inhibitor aminotriazole (AT), while its original background loh2 and wild-type plants are sensitive. Both, AT and PQ, which stimulate the intracellular formation of H₂O₂ or superoxide anions, respectively, trigger cell death in loh2 but do not lead to visible damage in atr7. To study gene expression during oxidative stress and ROS-induced programmed cell death, two platforms for multi-parallel quantitative real-time PCR (qRT-PCR) analysis of 217 antioxidant and 180 ROS marker genes were employed. The qRT-PCR analyses revealed AT- and PQ-induced expression of many ROS-responsive genes mainly in loh2, confirming that an oxidative burst plays a role in the activation of the cell death in this mutant. Some of the genes were specifically regulated by either AT or PQ, serving as markers for particular types of ROS. Genes significantly induced by both AT and PQ in loh2 included transcription factors (ANAC042/JUB1, ANAC102, DREB19, HSFA2, RRTF1, ZAT10, ZAT12, ethylene-responsive factors), signaling compounds, ferritins, alternative oxidases, and antioxidant enzymes. Many of these genes were upregulated in atr7 compared to loh2 under non-stress conditions at the first time point, indicating that higher basal levels of ROS and higher antioxidant capacity in atr7 are responsible for the enhanced tolerance to oxidative stress and suggesting a possible tolerance against multiple stresses of this mutant.

摘要

拟南芥 atr7 突变体对百草枯(PQ)或过氧化氢酶抑制剂氨基三唑(AT)诱导的氧化应激具有耐受性,而其原始背景 loh2 和野生型植物则对此敏感。AT 和 PQ 分别刺激细胞内 H₂O₂或超氧阴离子的形成,分别导致 loh2 中的细胞死亡,但不会导致 atr7 中出现可见的损伤。为了研究氧化应激和 ROS 诱导的程序性细胞死亡过程中的基因表达,我们使用了两种平台对 217 种抗氧化和 180 种 ROS 标记基因进行了多平行定量实时 PCR(qRT-PCR)分析。qRT-PCR 分析表明,ATR 和 PQ 诱导了许多 ROS 反应基因的表达,主要在 loh2 中,这证实了氧化爆发在该突变体中细胞死亡的激活中起作用。一些基因被 AT 或 PQ 特异性调节,可作为特定类型 ROS 的标记物。在 loh2 中,ATR 和 PQ 共同诱导的基因包括转录因子(ANAC042/JUB1、ANAC102、DREB19、HSFA2、RRTF1、ZAT10、ZAT12、乙烯响应因子)、信号化合物、铁蛋白、交替氧化酶和抗氧化酶。在非胁迫条件下,与 loh2 相比,许多这些基因在 atr7 中的第一个时间点就被上调,这表明 atr7 中 ROS 的基础水平较高和抗氧化能力较高,这使其对氧化应激具有更强的耐受性,并表明该突变体可能具有对多种胁迫的耐受性。

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