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Overexpression of a tonoplast Na+/H+ antiporter from the halophytic shrub Nitraria sibirica improved salt tolerance and root development in transgenic poplar

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Abstract

The sodium/proton exchanger (NHX) mediates Na+ and H+ countertransport in plants and plays an important role in regulating intracellular pH and maintaining ion and osmotic balance. Previously, an NHX1 orthologue was isolated from the halophyte Nitraria sibirica Pall (referred to as NsNHX1), and its role in enhancing salt tolerance of transgenic Arabidopsis was confirmed. To further analyse its features and functions, the 1311-bp sequence of the NsNHX1 promoter was cloned, and histochemical staining showed that β-glucuronidase (GUS) expression driven by the NsNHX1 promoter was strongly induced by abiotic stress and phytohormones, such as salt, drought, gibberellins, and methyl jasmonate, in turn indicating that NsNHX1 might participate in the regulation of various signalling pathways. To determine how NsNHX1 regulates salt tolerance in forestry trees, NsNHX1 was introduced into 84K poplar, and salt tolerance analysis of transgenic poplars showed that overexpression of NsNHX1 increased the overall biomass, survival rate, and plant height, and the contents of chlorophyll, proline and water, all of which are consequences of antioxidant enzyme activity under salt stress conditions. This showed that overexpression of NsNHX1 enhanced the salt tolerance of transgenic poplars as a result of NsNHX1-mediated Na+ compartmentalisation, more efficient photosynthesis, greater activity of antioxidant enzymes, and improved osmotic adjustment. Moreover, overexpression of NsNHX1 enhanced the root development of transgenic poplars; this resulted in increased biomass and height under normal and salt stress conditions, likely due to coupling between NsNHX1 and membrane proton pumps. These results provided a theoretical and experimental basis for further understanding the function and regulatory mechanism of NsNHX1, as well as its application for genetic improvement of forestry trees.

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References

  • Akama K, Shiraishi H, Ohta S, Nakamura K, Okada K, Shimura Y (1992) Efficient transformation of Arabidopsis thaliana: comparison of the efficiencies with various organs, plant ecotypes and Agrobacterium strain. Plant Cell Rep 12:7–11

    CAS  PubMed  Google Scholar 

  • Ballesterous E, Blumwald E, Donaire JP, Belver A (2010) Na+/H+ antiport activity in tonoplast vesicles isolated from sunflowers roots induced by NaCl stress. Physiol Plant 99(2):328–334

    Google Scholar 

  • Barkla BJ, Zingarelli L, Blumwald E, Smith JAC (1995) Tonoplast Na+/H+ antiport activity and its energization by the vacuolar H+-ATPase in the halophytic plant Mesembryanthemun crystallinum. L. Plant Physiol 109:549–556

    CAS  PubMed  PubMed Central  Google Scholar 

  • Blumwald E, Poole RJ (1985) Na+/H+ antiporter in isolated tonoplast vesicles from storage tissue of Beta vulgaris. Plant Physiol 78(1):163–167

    CAS  PubMed  PubMed Central  Google Scholar 

  • Blumwald E, Aharon GS, Apse MP (2000) Sodium transport in plant cells. Biochim Biophys Acta 1465(2000):140–151

    CAS  PubMed  Google Scholar 

  • Chakraborty K, Sairam RK, Bhattacharya R (2012) Differential expression of salt overly sensitive pathway genes determines salinity stress tolerance in Brassica genotypes. Plant Physiol Biochem 51:90–101

    CAS  PubMed  Google Scholar 

  • Chanroj S, Wang GY, Venema K, Zhang MW, Delwiche CF, Sze H (2012) Conserved and diversified gene families of monovalent cation/H+ antiporters from algae to flowering plants. Front Plant Sci 3(25):1–18

    Google Scholar 

  • Chen SL, Polle A (2010) Salinity tolerance of Populus. Plant Biol 12(2):317–333

    CAS  PubMed  Google Scholar 

  • Chen HT, Chen X, Gu HP, Wu BY, Zhang HM, Yuan XX, Cui XY (2014) GmHKT1;4, a novel soybean gene regulating Na+/K+ ratio in roots enhances salt tolerance in transgenic plants. Plant Growth Regul 73:299–308

    CAS  Google Scholar 

  • Cheng TL, Li HY, Wu HW, Liu ZX, Wu X, Yang S, Zhang HX, Yang XY (2015) Comparison on Osmotica accumulation of different salt-tolerant plants under salt stress. For Res 28(6):826–832

    Google Scholar 

  • Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743

    CAS  PubMed  Google Scholar 

  • Elena D (2012) Brief approach of plant-insect herbivores physiological interactions. J Hortic For Biotechnol 16(2):17–31

    Google Scholar 

  • Guan B, Hu YZ, Zeng YL, Wang Y, Zhang FC (2011) Molecular characterization and functional analysis of a tonoplast Na+/H+ antiporter gene (HcNHX1) from Halostachys caspica. Mol Biol Rep 38(3):1889–1899

    CAS  PubMed  Google Scholar 

  • He CX, Yan JQ, Shen GX, Fu LH, Holaday A, Auld D, Blumwald E, Zhang H (2005) Expression of an Arabidopsis vacuolar sodium/proton antiporter gene in cotton improves photosynthetic performance under salt conditions and increases fiber yield in the field. Plant Cell Physiol 46(11):1848–1854

    CAS  PubMed  Google Scholar 

  • Jha A, Joshi M, Yadav NS, Agarwal PK, Jha B (2011) Cloning and characterization of the Salicornia brachiata Na+/H+ antiporter gene SbNHX1 and its expression by abiotic stress. Mol Biol Rep 38:1965–1973

    CAS  PubMed  Google Scholar 

  • Li TX, Zhang Y, Liu H, Wu YT, Li WB, Zhang HX (2010) Stable expression of Arabidopsis vacuolar Na+/H+ antiporter gene AtNHX1, and salt tolerance in transgenic soybean for over six generations. Sci Bull 55(12):1127–1134

    CAS  Google Scholar 

  • Li NN, Wang X, Ma BJ, Du C, Zheng LL, Wang YC (2017) Expression of a Na+/H+ antiporter RtNHX1 from a recretohalophyte Reaumuria trigyna improved salt tolerance of transgenic Arabidopsis thaliana. J Plant Physiol 218:109–120

    CAS  PubMed  Google Scholar 

  • Liang D, Wang JH, Liu D, Wang CL, Shi XW, Wu Q, Zheng JJ, Feng G (2016) The effect on germination and seedlings of wheat under different salt stresses. Agric Sci Technol 17(11):2564–2568

    Google Scholar 

  • Liu L, Zeng YL, Zhang FC (2012) Regulation of rice (Oryza sativa) NHX1 gene: role of its promoter and C-terminal. Acta Botan Boreali-Occiden Sin 32(7):1295–1303

    CAS  Google Scholar 

  • Ma XL, Zhang Q, Shi HZ, Zhu JK, Zhao YX, Ma CL, Zhang H (2004) Molecular cloning and different expression of a tonoplast Na+/H+ antiporter gene in Suaeda salsa under salt stress. Biol Plant 48(2):219–225

    CAS  Google Scholar 

  • Nie WX, Xu L, Yu BJ (2015) A putative soybean GmsSOS1 confers enhanced salt tolerance to transgenic Arabidopsis sos1-1 mutant. Protoplasma 252(1):127–134

    CAS  PubMed  Google Scholar 

  • Niu X, Bressan RA, Hasegawa PM, Pardo JM (1995) Ion homeostasis in NaCl stress environments. Plant Physiol 109:735–742

    CAS  PubMed  PubMed Central  Google Scholar 

  • Olias R, Eljakaoui Z, Li J, Morales PAD, Marin-manzano MC, Pardo JM, Belver A (2009) The plasma membrane Na+/H+ antiporter SOS1 is essential for salt tolerance in tomato and affects the partitioning of Na+ between plant organs. Plant Cell Environ 32(7):904–916

    CAS  PubMed  Google Scholar 

  • Pehlivan N, Sun L, Jarrett P, Yang XJ, Mishra N, Chen L, Kadioglu A, Shen GX, Zhang H (2016) Co-overexpressing a plasma membrane and a vacuolar membrane sodium/proton antiporter significantly improves salt tolerance in transgenic Arabidopsis plants. Plant Cell Physiol 57(5):1069–1084

    CAS  PubMed  PubMed Central  Google Scholar 

  • Polle A, Chen SL (2014) On the salty side of life: molecular, physiological and anatomical adaptation and acclimation of trees to extreme habitats. Plant Cell Environ 38(9):1794–1816

    PubMed  Google Scholar 

  • Poulsen LR, Palmgren MG, López-Marqués RL (2016) Transient expression of P-type ATPases in tobacco epidermal cells. Methods Mol Biol 1377:383–393

    CAS  PubMed  Google Scholar 

  • Qiao WH, Zhao XY, Li W, Luo Y, Zhang XS (2007) Overexpression of AeNHX1, a root-specific vacuolar Na+/H+ antiporter from Agropyron elongatum, confers salt tolerance to Arabidopsis and Festuca plants. Plant Cell Rep 26(9):1663–1672

    CAS  PubMed  Google Scholar 

  • Qiao GR, Zhuo RY, Liu MY, Jiang J, Li HY, Qiu WM, Pan LY, Lin S, Zhang XG, Sun ZX (2011) Over-expression of the Arabidopsis Na+/H+ antiporter gene in Populus deltoides CL×P. euramericana CL “NL895” enhances its salt tolerance. Acta Physiol Plant 33:691–696

    CAS  Google Scholar 

  • Sahoo DP, Kumar S, Mishra S, Kobayashi Y, Panda SK, Sahoo L (2016) Enhanced salinity tolerance in transgenic mungbean overexpressing Arabidopsis antiporter (NHX1) gene. Mol Breed 36(10):144–158

    Google Scholar 

  • Shi HZ, Zhu JK (2002) Regulation of expression of the tonoplast Na+/H+ antiporter gene AtNHX1 by salt stress and abscisic acid. Plant Mol Biol 50:543–550

    CAS  PubMed  Google Scholar 

  • Tang RJ, Yang Y, Yang L, Liu H, Wang CT, Yu MM, Gao XS, Zhang HX (2014) Poplar calcineurin B-like proteins PtCBL10A and PtCBL10B regulate shoot salt tolerance through interaction with PtSOS2 in the vacuolar membrane. Plant Cell Environ 37:573–588

    CAS  PubMed  Google Scholar 

  • Verma D, Singla-Pareek SL, Rajagopal D, Reddy MK, Sopory SK (2007) Functional validation of a novel isoform of Na+/H+ antiporter from Pennisetum glaucum for enhancing salinity tolerance in rice. J Biosci 32(3):1–7

    Google Scholar 

  • Wang L, Ma YK, Li NN, Zhang WB, Mao HY, Lin XF (2016) Isolation and characterization of a tonoplast Na+/H+ antiporter from the halophyte Nitraria sibirica. Biol Plant 60(1):113–122

    CAS  Google Scholar 

  • Wu SJ, Ding L, Zhu JK (1996) SOS1, a genetic locus essential for salt tolerance and potassium acquisition. Plant Cell 8(4):617–627

    CAS  PubMed  PubMed Central  Google Scholar 

  • Wu CX, Gao XH, Kong XQ, Zhao YX, Zhang H (2009) Molecular cloning and functional analysis of a Na+/H+ antiporter gene ThNHX1 from a halophytic plant Thellungiella halophila. Plant Mol Biol Report 27(1):1–12

    CAS  Google Scholar 

  • Wu GQ, Xi JJ, Wang Q, Bao AK, Ma Q, Zhang JL, Wang SM (2011) The ZxNHX gene encoding tonoplast Na+/H+ antiporter from the xerophyte Zygophyllum xanthoxylum plays important roles in response to salt and drought. J Plant Physiol 168(8):758–767

    CAS  PubMed  Google Scholar 

  • Xu M, Chen CH, Cai H, Wu L (2018) Overexpression of PeHKT1;1 improves salt tolerance in Populus. Genes 9(10):475–489

    PubMed Central  Google Scholar 

  • Yamaguchi T, Blumwald E (2005) Developing salt-tolerant crop plants: challenges and opportunities. Trends Plant Sci 10(12):615–620

    CAS  PubMed  Google Scholar 

  • Yang YL, Shi RX, Wei XL, Fan Q, An LZ (2010a) Effect of salinity on antioxidant enzymes in calli of the halophyte Nitraria tangutorum Bobr. Plant Cell Tissue Organ Cult 102:387–395

    CAS  Google Scholar 

  • Yang YL, Wei XL, Shi RX, Fan Q, An LZ (2010b) Salinity-induced physiological modification in the callus from halophyte Nitraria tangutorum Bobr. J Plant Growth Regul 29:465–476

    CAS  Google Scholar 

  • Zhang HX, Blumwald E (2001) Transgenic salt-tolerant tomato plants accumulate salt in foliage but not in fruit. Nat Biotechnol 19(8):765–768

    CAS  PubMed  Google Scholar 

  • Zhang HX, Hodson JN, Williams JP, Blumwald E (2001) Engineering salt-tolerant Brassica plant: characterization of yield and seed oil quality in transgenic plants with increase vacuolar sodium accumulation. Proc Natl Acad Sci U S A 98:12832–12836

    CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang XY, Zhu Y, Wu HH, Guo HW (2016) Post-transcriptional gene silencing in plants: a double-edged sword. Sci China Life Sci 59:271–276

    PubMed  Google Scholar 

  • Zhu JK (2001) Plant salt tolerance. Trends Plant Sci 6:66–71

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We would like to thank Professor Haifeng Yang in Inner Mongolia Agricultural University, China, for kindly providing 84K poplar (Populus alba × Populus glandulosa) and Professor Nakamura Kenzo in Chubu University for kindly providing pBI101-35::Gus-Hm vector.

Funding

This work was supported by the National Natural Science Foundation of China (31860216, 31560065, 31660213), the Inner Mongolia Autonomous Region Science and Technology Innovation Guidance Award Fund (2017), and the Inner Mongolia Natural Science Foundation (2017MS0329).

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Author notes

  1. Xin Geng, Shouye Chen and E. Yilan contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory of Forage and Endemic Crop Biotechnology, Ministry Education, School of Life Sciences, Inner Mongolia University, 235 Daxuexi Road, Hohhot, 010021, China

    Xin Geng, Shouye Chen, E. Yilan, Huiping Mao, Alatan qiqige, Yingchun Wang, Zhi Qi & Xiaofei Lin

  2. College of Forestry, Inner Mongolia Agricultural University, Hohhot, 010019, China

    Wenbo Zhang

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  1. Xin Geng
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Contributions

Conceived and designed the experiments: Xiaofei Lin.

Performed the experiments and analysed the data: Xin Geng, Shouye Chen, Yilan E, Wenbo Zhang, Huiping Mao, Alatan qiqige, Yingchun Wang, Zhi Qi, and Xiaofei Lin.

Wrote the paper: Xin Geng, Xiaofei Lin, and Shouye Chen.

Statement: Xin Geng, Shouye Chen, and Yilan E contributed equally to this paper.

Corresponding author

Correspondence to Xiaofei Lin.

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The authors declare that they have no conflict of interest.

Data archiving statement

NsNHX1 sequences and information of Nitraria sibirica were deposited in the NCBI database (accession number: AB859847, accessible with the following link: https://www.ncbi.nlm.nih.gov/nuccore/820687834).

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Communicated by A. De La Torre

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Geng, X., Chen, S., Yilan, E. et al. Overexpression of a tonoplast Na+/H+ antiporter from the halophytic shrub Nitraria sibirica improved salt tolerance and root development in transgenic poplar. Tree Genetics & Genomes 16, 81 (2020). https://doi.org/10.1007/s11295-020-01475-7

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