J Integr Plant Biol. ›› 2008, Vol. 50 ›› Issue (2): 157-167.DOI: 10.1111/j.1744-7909.2007.00609.x

Special Issue: Abiotic Stresses Plant Signal Transduction

• Stress & Phytochemistry • Previous Articles     Next Articles

Excessive Copper Induces the Production of ROS, which is Mediated by Phospholipase D, NADPH Oxidase and Antioxidant Systems

Zhong-Lian Yu, Jin-Guang Zhang, Xue-Chen Wang and Jia Chen   

  • Received:2007-01-08 Accepted:2007-06-09

Abstract: Tobacco BY-2 suspension cells were used to study the chemical damage and its associated mechanisms caused by Cu2+. Treatment with 100 μmol/L Cu2+ generated a large amount of H2O2 and thiobarbituric acid-reactive substances (TBARS) in cells. Using phospholipase D (PLD) specific inhibitor (1-butanol) or phosphatidic acid (PA), we demonstrated that PLD plays an important role in the generation of H2O2 and TBARS. Semi-quantitative reverse-transcriptase polymerase chain reaction and enzyme activity assays with wild type and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase-overexpressing BY-2 cells revealed that PLD and PA are the key factors leading to NADPH oxidase activation, which is responsible for H2O2 and TBARS production induced by Cu2+. Moreover, the content of ascorbic acid (AsA), an effective antioxidant, was sharply reduced in BY-2 cells exposed to excessive Cu2+. Furthermore, a significant downregulation of the enzymes of AsA biosynthesis and the antioxidant system was found. This evidence suggests that excessive Cu2+-elevated reactive oxygen species (ROS) production is caused by upregulated PLD that elevates the activity of NADPH oxidase and its collapsed antioxidant systems that scavenges ROS.

Key words: NADPH oxidase, phospholipase D, ROS, Cu2+, ascorbic acid.

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