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Halophytic NHXs confer salt tolerance by altering cytosolic and vacuolar K+ and Na+ in Arabidopsis root cell

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Abstract

While the role of the vacuolar NHX Na+/H+ exchangers in plant salt tolerance has been demonstrated on numerous occasions, their control over cytosolic ionic relations has never been functionally analysed in the context of subcellular Na+ and K+ homeostasis. In this work, PutNHX1 and SeNHX1 were cloned from halophytes Puccinellia tenuiflora and Salicornia europaea and transiently expressed in Arabidopsis wild type Col-0 and the nhx1 mutant. Phylogentic analysis, topological prediction, analysis of evolutionary conservation, the topology structure and analysis of hydrophobic or polar regions of PutNHX1 and SeNHX1 indicated that they are unique tonoplast Na+/H+ antiporters with characteristics for salt tolerance. As a part of the functional assessment, cytosolic and vacuolar Na+ and K+ in different root tissues and ion fluxes from root mature zone of Col-0, nhx1 and their transgenic lines were measured. Transgenic lines sequestered large quantity of Na+ into root cell vacuoles and also promoted high cytosolic and vacuolar K+ accumulation. Expression of PutNHX1 and SeNHX1 led to significant transient root Na+ uptake in the four transgenic lines upon recovery from salt treatment. In contrast, the nhx1 mutant maintained a prolonged Na+ efflux and the nhx1:PutNHX1 and nhx1:SeNHX1 lines started to actively pump Na+ out of the cell. Overall, our findings suggest that PutNHX1 and SeNHX1 improve Na+ sequestration in the vacuole and K+ retention in the cytosol and vacuole of root cells of Arabidopsis, and that they interact with other regulatory mechanisms to provide a highly orchestrated regulation of ionic relations among intracellular cell compartments.

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Acknowledgements

We thank Dr Christopher Grefen for providing the pUB-DEST and pUBC-RFP-DEST vectors, Dr Vivien Rolland and A/Prof Spencer Whitney for P19 strain and Guang Chen for constructing the phylogenetic tree. This project was supported by an Australian Research Council (ARC) DECRA award (DE140101143) and a Chinese 1000-Plan project to Zhong-Hua Chen. Xiaohui Liu is a recipient of China Scholarship Council (CSC) award. Gang Wang and Jing Ji are supported by National Natural Science Foundation of China projects (31271419 and 31271793).

Authors' contributions

Z.H.C. X.L. and G.W. conceived the original research plans and designed the experiments; Z.H.C. P.H. J.J. A.S. and S.S. and supervised the experiments; X.L. and S.C. performed most of the experiments with assistance from F.W. F.D. M.M. and M.Z.; X.L. S.C. and Z.H.C. analysed the data; Z.H.C. X.L. P.H. and S.S. wrote the article with contributions of all the authors.

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    1. School of Science and Health, Hawkesbury Institute for the Environment, Western Sydney University, Penrith, NSW, 2751, Australia

      Xiaohui Liu, Shenguan Cai, Michelle Mak, Paul Holford, Anya Salih & Zhong-Hua Chen

    2. School of Environmental Science and Engineering, Tianjin University, Tianjin, 300072, China

      Xiaohui Liu, Gang Wang, Fabao Dong & Jing Ji

    3. School of Land and Food, University of Tasmania, Hobart, TAS, 7005, Australia

      Feifei Wang, Meixue Zhou & Sergey Shabala

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    Liu, X., Cai, S., Wang, G. et al. Halophytic NHXs confer salt tolerance by altering cytosolic and vacuolar K+ and Na+ in Arabidopsis root cell. Plant Growth Regul 82, 333–351 (2017). https://doi.org/10.1007/s10725-017-0262-7

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