Metal-specific and NADPH oxidase dependent changes in lipoxygenase and NADPH oxidase gene expression in Arabidopsis thaliana exposed to cadmium or excess copper
Tony Remans A B , Kelly Opdenakker A , Karen Smeets A , Dennis Mathijsen A , Jaco Vangronsveld A and Ann Cuypers AA Centre for Environmental Sciences, Hasselt University, Campus Diepenbeek, Agoralaan Building D, 3590 Diepenbeek, Belgium.
B Corresponding author. Email: tony.remans@uhasselt.be
Functional Plant Biology 37(6) 532-544 https://doi.org/10.1071/FP09194
Submitted: 24 July 2009 Accepted: 2 March 2010 Published: 20 May 2010
Abstract
Reactive oxygen species produced by NADPH oxidase and oxylipins derived from lipoxygenase activity can signal various stress conditions and have been implicated when plants are exposed to heavy metals. Transcriptional profiling of the 10 NADPH oxidase and 6 lipoxygenase genes was performed after exposure of Arabidopsis thaliana wild-type and NADPH oxidase mutants to 5 µM CdSO4 or 2 µM CuSO4 for 24 h. Under these short exposures to environmentally realistic concentrations of Cd or Cu, plants modulate signalling networks that regulate the onset of adaptive responses. Metal-specific NADPH oxidase genes were upregulated by Cd but downregulated by Cu, and metal-specific lipoxygenase gene expression was observed only after Cu exposure. Genes that are responsive to both metals were upregulated and may be responsive to general oxidative stress. For all metal-responsive genes except RBOHD, distinct responses were observed between leaves and roots, which may be due to different stress intensities and signalling mechanisms. Mutation of NADPH oxidase genes had opposing effects on gene expression after Cd or Cu exposure. Upregulation of LOX1 and LOX6 in the roots after exposure to Cd depended on NADPH oxidase gene expression, whereas LOX3 and LOX6 expression was induced more strongly in NADPH oxidase mutants after Cu exposure. Furthermore, NADPH oxidases regulated their own expression level and that of other members of the gene family when exposed to Cd or Cu. The results suggest interplay between reactive oxygen species and oxylipin signalling under Cd or Cu stress, and are useful as a basis for genetic studies to unravel metal-specific signalling mechanisms.
Additional keywords: abiotic stress, gene family, heavy metals, hydroponic growth, hydroponics, normalisation, OXI1, oxidative stress, qPCR, real-time PCR, real-time RT-PCR, respiratory burst oxidase homologue, signaling, signalling, superoxide dismutase.
Acknowledgements
This work was supported by the Research Foundation-Flanders (FWO) (project G.0436.06 and G.0807.09). Tony Remans is a post-doctoral researcher of the FWO. Additional funding came from Hasselt University through BOF (Bijzonder Onderzoeksfonds) projects of Hasselt University BOF08G01, BOFN0705 and Methusalem project (08M03VGRJ). The authors gratefully acknowledge Dr Miguel Torres (Centro de Biotecnología y Genómica de Plantas, Universidad Politécnica de Madrid, Madrid 28223, Spain) for providing NADPH oxidase mutant seed, and Professor R. Carleer and his staff for the element determinations. Carine Put and Ann Wijgaerts are acknowledged for their skilful technical assistance.
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