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CAX3 (cation/proton exchanger) mediates a Cd tolerance by decreasing ROS through Ca elevation in Arabidopsis

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Over-expression of CAX3 encoding a cation/proton exchanger enhances Cd tolerance by decreasing ROS (Reactive Oxygen Species) through activating anti-oxidative enzymes via elevation of Ca level in Arabidopsis

Abstract

CAXs (cation/proton exchangers) are involved in the sequestration of cations such as Mn, Li, and Cd, as well as Ca, from cytosol into the vacuole using proton gradients. In addition, it has been reported that CAX1, 2 and 4 are involved in Cd tolerance. Interestingly, it has been reported that CAX3 expressions were enhanced by Cd in Cd-tolerant transgenic plants expressing Hb1 (hemoglobin 1) or UBC1 (Ub-conjugating enzyme 1). Therefore, to investigate whether CAX3 plays a role in increasing Cd tolerance, CAX3 of Arabidopsis and tobacco were over-expressed in Arabidopsis thaliana. Compared to control plants, both transgenic plants displayed an increase in Cd tolerance, no change in Cd accumulation, and enhanced Ca levels. In support of these, AtCAX3-Arabidopsis showed no change in expressions of Cd transporters, but reduced expressions of Ca exporters and lower rate of Ca efflux. By contrast, atcax3 knockout Arabidopsis exhibited a reduced Cd tolerance, while the Cd level was not altered. The expression of Δ90-AtCAX3 (deletion of autoinhibitory domain) increased Cd and Ca tolerance in yeast, while AtCAX3 expression did not. Interestingly, less accumulation of ROS (H2O2 and O2) was observed in CAX3-expressing transgenic plants and was accompanied with higher antioxidant enzyme activities (SOD, CAT, GR). Taken together, CAX3 over-expression may enhance Cd tolerance by decreasing Cd-induced ROS production by activating antioxidant enzymes and by intervening the positive feedback circuit between ROS generation and Cd-induced spikes of cytoplasmic Ca.

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Acknowledgements

We thank Prof. K. D. Hirschi, Children’s Nutrition Research Center (CNRC), Baylor College of Medicine, Houston, Texas, USA for providing the atcax3 knockout mutant seeds, yeast K667, vector pRS306, constructs pRS306:AtCAX3 and pRS306:Δ90-AtCAX3, and valuable comments and discussions.

Funding

This work was supported by the Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries (IPET) through the Agri-Bio industry Technology Development Program, funded by the Ministry of Agriculture, Food and Rural Affairs (MAFRA) (IPET 319107–4), Korea.

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SH conceived the project and wrote the article with contributions from all the authors; MM and RB performed the majority of the experiments; DK performed some, more minor, parts of the experiments. MM and RB contributed equally to this work.

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Correspondence to Seongbin Hwang.

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Accession numbers

Sequence data from this article can be found in the TAIR (https://www.arabidopsis.org) data libraries under the following accession numbers: AtActin2 (AT3G18780), AtCAX1 (AT2G38170), AtCAX2 (AT3G13320), AtCAX3 (AT3G51860), AtCAX4 (AT5G01490), AtHMA2 (AT4G30110), AtHMA4 (AT2G19110), AtNRAMP3 (AT2G23150), AtPDR8 (AT1G59870), AtABCC1 (AT1G30400), AtIRT1 (AT4G19690), AtACA8 (AT5G56330), AtACA10 (AT4G29900), AtGLR3.3 (AT1G42540), AtMCA1 (AT4G35920), AtCNGC10 (AT1G01340), NtCAX3 (MK875639).

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Modareszadeh, M., Bahmani, R., Kim, D. et al. CAX3 (cation/proton exchanger) mediates a Cd tolerance by decreasing ROS through Ca elevation in Arabidopsis. Plant Mol Biol 105, 115–132 (2021). https://doi.org/10.1007/s11103-020-01072-1

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  • DOI: https://doi.org/10.1007/s11103-020-01072-1

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