Biologia plantarum 64: 50-57, 2020 | DOI: 10.32615/bp.2019.126

Overexpression of Iris lactea tonoplast Na+/H+ antiporter gene IlNHX confers improved salt tolerance in tobacco

Q. GUO, X.X. TIAN, P.C. MAO, L. MENG*
Beijing Research and Development Center for Grass and Environment,
Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, P.R. China

Sodium cation compartmentalization into vacuoles is one of the effective strategies for adaptation of halophytes to saline environments. Tonoplast Na+/H+ antiporter (NHX) is involved in Na+ sequestration into vacuoles under salt stress. However, the function of NHX in halophyte Iris lactea is still unclear. In this study, a significant positive correlation was observed between Na+ accumulations and IlNHX expression in tissues under 0 - 200 mM NaCl, indicating IlNHX might be responsible for Na+ accumulation of I. lactea under salt stress. More important, IlNHX was specifically localized to the tonoplast. Transgenic tobacco expressing IlNHX grew better and showed higher tolerance to 200 mM NaCl than respective wild type (WT). Compared to WT, transgenic tobacco accumulated more Na+ and K+ and maintained higher K+/Na+ ratios in tissues, accompanied by the reduction of chlorophyll loss and lipid peroxidation in the presence of NaCl. Moreover, transgenic tobacco exhibited markedly higher vacuolar H+-ATPase (V-ATPase) activity relative to WT when subjected to salt stress. The findings suggest that transgenic plants overexpressing IlNHX could compartmentalize more Na+ into vacuoles in tobacco via enhanced V-ATPase activity, which further contributes to maintaining K+ and Na+ homeostasis, to improved photosynthesis, and to protection of cell membrane integrity under salt stress.

Keywords: K+/Na+ ratios, Na+ compartmentalization, Nicotiana tabacum, tonoplast Na+/H+ antiporter, vacuolar H+-ATPase.

Received: June 16, 2019; Revised: October 7, 2019; Accepted: October 15, 2019; Prepublished online: January 27, 2020; Published online: January 17, 2020Show citation

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GUO, Q., TIAN, X.X., MAO, P.C., & MENG, L. (2020). Overexpression of Iris lactea tonoplast Na+/H+ antiporter gene IlNHX confers improved salt tolerance in tobacco. Biologia plantarum, 64, 50-57. doi: 10.32615/bp.2019.126.
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    References

    1. Akbari, M., Mahna, N., Ramesh, K., Bandehagh, A., Mazzuca, S.: Ion homeostasis, osmoregulation, and physiological changes in the roots and leaves of pistachio rootstocks in response to salinity. - Protoplasma 255: 1349-1362, 2018. Go to original source...
    2. Apse, M.P., Aharon, G.S., Snedden, W.A., Blumwald, E.: Salt tolerance conferred by overexpression of a vacuolar Na+/H+ antiport in Arabidopsis. - Science 285: 1256-1258, 1999. Go to original source...
    3. Apse, M.P., Blumwald, E.: Na+ transport in plants. - FEBS Lett. 581: 2247-2254, 2007. Go to original source...
    4. Bai, W.B., Li, P.F.: [Effects of salt stress on growth of and absorption and transportation of K+ and Na+ in Iris lactea var. chinensis.] - Soils 37: 415-420, 2005. [In Chinese]
    5. Bao, A.K., Du, B.Q., Touil, L., Kang, P., Wang, Q.L., Wang, S.M.: Co-expression of tonoplast cation/H+ antiporter and H+-pyrophosphatase from xerophyte Zygophyllum xanthoxylum improves alfalfa plant growth under salinity, drought and field conditions. - Plant Biotechnol. J. 14: 964-975, 2016. Go to original source...
    6. Barragán, V., Leidi, E.O., Andrés, Z., Rubio, L., De Luca, A., Fernández, J.A., Cubero, B., Pardo, J.M.: Ion exchangers NHX1 and NHX2 mediate active potassium uptake into vacuoles to regulate cell turgor and stomatal function in Arabidopsis. - Plant Cell 24: 1127-1142, 2012. Go to original source...
    7. Bassil, E., Blumwald, E.: The ins and outs of intracellular ion homeostasis: NHX-type cation/H+ transporters. - Curr. Opin. Plant Biol. 22: 1-6, 2014. Go to original source...
    8. Bassil, E., Tajima, H., Liang, Y.C., Ohto, M.A., Ushijima, K., Nakano, R., Esumi, T., Coku, A., Belmonte, M., Blumwald, E.: The Arabidopsis Na+/H+ antiporters NHX1 and NHX2 control vacuolar pH and K+ homeostasis to regulate growth, flower development, and reproduction. - Plant Cell 23: 3482-3497, 2011. Go to original source...
    9. Blumwald, E., Aharon, G.S., Apse, M.P.: Sodium transport in plant cells. - Biochim. biophys. Acta 1465: 140-151, 2000. Go to original source...
    10. Dhindsa, R.S., Plumb-Dhindsa, P., Thorpe, T.A.: Leaf senescence: correlated with increased levels of membrane permeability and lipid peroxidation, and decreased levels of superoxide dismutase and catalase. - J. exp. Bot. 32: 93-101, 1981. Go to original source...
    11. Fan, W., Deng, G., Wang, H., Zhang, H., Zhang, P.: Elevated compartmentalization of Na+ into vacuoles improves salt and cold stress tolerance in sweet potato (Ipomoea batatas). - Physiol. Plant. 154: 560-571, 2015. Go to original source...
    12. Flowers, T.J., Colmer, T.D.: Salinity tolerance in halophytes. -New Phytol. 179: 945-963, 2008. Go to original source...
    13. Gaxiola, R.A., Palmgren, M.G., Schumacher, K.: Plant proton pumps. - FEBS Lett. 581: 2204-2214, 2007. Go to original source...
    14. Gaxiola, R.A., Rao, R., Sherman, A., Grisafi, P., Alper, S.L., Fink, G.R.: The Arabidopsis thaliana proton transporters, AtNhx1 and Avp1, can function in cation detoxification in yeast. -Proc. nat. Acad. Sci. USA 96: 1480-1485, 1999. Go to original source...
    15. Gouiaa, S., Khoudi, H.: Co-expression of vacuolar Na+/H+ antiporter and H+-pyrophosphatase with an IRES-mediated dicistronic vector improves salinity tolerance and enhances potassium biofortification of tomato. -Phytochemistry 117: 537-546, 2015. Go to original source...
    16. Guo, Q., Meng, L., Humphreys, M.W., Scullion, J., Mur, L.A.J.: Expression of FlHMA3, a P1B2-ATPase from Festulolium loliaceum, correlates with response to cadmium stress. - Plant Physiol. Biochem. 112: 270-277, 2017b. Go to original source...
    17. Guo, Q., Meng, L., Li, S.S., Zhang, L., Mao, P.C., Tian, X.X.: [Cloning of Iris lactea NHX and analysis of gene expression.] - Plant Physiol. J. 51: 2006-2012, 2015a. [In Chinese].
    18. Guo, Q., Meng, L., Mao, P.C., Tian, X.X.: An assessment of Agropyron cristatum tolerance to cadmium contaminated soil. - Biol. Plant. 58: 174-178, 2014. Go to original source...
    19. Guo, Q., Meng, L., Mao, P.C., Tian, X.X.: Salt tolerance in two tall wheatgrass species is associated with selective capacity for K+ over Na+. - Acta Physiol. Plant. 37: 1708, 2015b. Go to original source...
    20. Guo, Q., Meng, L., Zhang, Y.N., Mao, P.C., Tian, X.X., Li, S.S., Zhang, L.: Antioxidative systems, metal ion homeostasis and cadmium distribution in Iris lactea exposed to cadmium stress. - Ecotox. environ. Safety 139: 50-55, 2017a. Go to original source...
    21. Horsch, R.B., Fry, J.E., Hoffmann, N.L., Eichholtz, D., Rogers, S.G., Fraley, R.T.: A simple and general method for transferring genes into plants. - Science 227: 1229-1232, 1985.
    22. Kumari, P.H., Kumar, S.A., Sivan, P., Katam, R.: Overexpression of a plasma membrane bound Na+/H+ antiporter-like protein (SbNHXLP) confers salt tolerance and improves fruit yield in tomato by maintaining ion homeostasis. - Front. Plant Sci. 7: 2027, 2017.
    23. Leidi, E.O., Barragan, V., Rubio, L., El-Hamdaoui, A., Ruiz, M.T., Cubero, B., Fernandez, J.A., Bressan, R.A., Hasegawa, P.M., Quintero, F.J., Pardo, J.M.: The AtNHX1 exchanger mediates potassium compartmentation in vacuoles of transgenic tomato. - Plant J. 61: 495-506, 2010. Go to original source...
    24. Ma, Q., Li, Y.X., Yuan, H.J., Hu, J., Wei, L., Bao, A.K., Zhang, J.L., Wang, S.M.: ZxSOS1 is essential for long-distance transport and spatial distribution of Na+ and K+ in the xerophyte Zygophyllum xanthoxylum. - Plant Soil 374: 661-676, 2014. Go to original source...
    25. Mao, P.C., Tian, X.X., Meng, L.: [Evaluation of salt tolerance for 16 Iris lactea var. chinensis accessions at seeding stage.] - Pratacult. Sci. 30: 35-43, 2013. [In Chinese]
    26. Mishra, S., Behura, R., Awasthi, J.P., Dey, M., Sahoo, D., Bhowmik, S.S.D., Panda, S.K., Sahoo, L.: Ectopic overexpression of a mung bean vacuolar Na+/H+ antiporter gene (VrNHX1) leads to increased salinity stress tolerance in transgenic Vigna unguiculata L. Walp. - Mol. Breed. 34: 1345-1359, 2014. Go to original source...
    27. Munns, R., Tester, M.: Mechanisms of salinity tolerance. - Annu. Rev. Plant Biol. 59: 651-681, 2008. Go to original source...
    28. Murguía, J.R., Bellés, J.M., Serrano, R.: A salt-sensitive 3'(2'),5'-bisphosphate nucleotidase involved in sulfate activation. - Science 267: 232-234, 1995. Go to original source...
    29. Pardo, J.M., Cubero, B., Leidi, E.O., Quintero, F.J.: Alkali cation exchangers: roles in cellular homeostasis and stress tolerance. - J. exp. Bot. 57: 1181-1199, 2006. Go to original source...
    30. Reguera, M., Bassil, E., Blumwald, E.: Intracellular NHX-type cation/H+ antiporters in plants. - Mol. Plants 7: 261-263, 2014. Go to original source...
    31. Shabala, S.: Learning from halophytes: physiological basis and strategies to improve abiotic stress tolerance in crops. - Ann. Bot. 112: 1209-1221, 2013. Go to original source...
    32. Shi, H.H., Zhu, J.K.: Regulation of expression of the vacuolar Na+/H+ antiporter gene AtNHX1 by salt stress and abscisic acid. - Plant mol. Biol. 50: 543-550, 2002. Go to original source...
    33. Tsunekawa, K., Shijuku, T., Hayashimoto, M., Kojima, Y., Onai, K., Morishita, M., Ishiura, M., Kuroda, T., Nakamura, T., Kobayashi, H., Sato, M., Toyooka, K., Matsuoka, K., Omata, T., Uozumi, N.: Identification and characterization of the Na+/H+antiporter Nhas3 from the thylakoid membrane of Synechocystis sp. PCC 6803. - J. biol. Chem. 284: 16513-16521, 2009. Go to original source...
    34. Wang, B.S., Ratajczak, R., Zhang, J.H.: Activity, amount and subunit composition of vacuolar-type H+-ATPase and H+-PPase in wheat roots under severe NaCl stress. - J. Plant Physiol. 157: 109-116, 2000. Go to original source...
    35. Wu, G.Q., Liang, N., Feng, R.J., Zhang, J.J.: Evaluation of salinity tolerance in seedlings of sugar beet (Beta vulgaris L.) cultivars using proline, soluble sugars and cation accumulation criteria. - Acta. Physiol. Plant. 35: 2665-2674, 2013. Go to original source...
    36. Wu, G.Q., Feng, R.J., Wang, S.M., Wang, C.M., Bao, A.K., Wei, L., Yuan, H.J.: Co-expression of xerophyte Zygophyllum xanthoxylum ZxNHX and ZxVP1-1 confers enhanced salinity tolerance in chimeric sugar beet (Beta vulgaris L.). -Front. Plant Sci. 6: 581, 2015. Go to original source...
    37. Xia, T., Apse, M.P., Aharon, G.S., Blumwald, E.: Identification and characterization of a NaCl-inducible vacuolar Na+/H+ antiporter in Beta vulgaris. - Physiol. Plant. 116: 206-212, 2002. Go to original source...
    38. Yamaguchi, T., Fukuda-Tanaka, S., Inagaki, Y., Saito, N., Yonekura-Sakakibara, K., Tanaka, Y., Kusumi, T., Iida, S.: Genes encoding the vacuolar Na+/H+ exchanger and flower coloration. - Plant Cell Physiol. 42: 451-461, 2001. Go to original source...
    39. Yamaguchi, T., Hamamoto, S., Uozumi, N.: Sodium transport system in plant cells. - Front. Plant Sci. 4: 410, 2013. Go to original source...
    40. Ye, C.Y., Zhang, H.C., Chen, J.H., Xia, X.L., Yin, W.L.: Molecular characterization of putative vacuolar NHX-type Na+/H+ exchanger genes from the salt-resistant tree Populus euphratica. - Physiol. Plant. 137: 166-174, 2009. Go to original source...
    41. Yeo, A.R.: Salt tolerance in the halophyte Suaeda maritime (L.) Dum.: intracellular compartmentation of ions. -J. exp. Bot. 32:487-497, 1981. Go to original source...
    42. Zhang, H.X., Hodson, J.N., Williams, J.P., Blumwald, E.: Engineering salt-tolerant Brassica plants: characterization of yield and seed oil quality in transgenic plants with increased vacuolar sodium accumulation. - Proc. nat. Acad. Sci. USA 98: 12832-12836, 2001. Go to original source...
    43. Zhang, Y.M., Zhang, H.M., Liu, Z.H., Li, H.C., Guo, X.L., Li, G.L.: The wheat NHX antiporter gene TaNHX2 confers salt tolerance in transgenic alfalfa by increasing the retention capacity of intracellular potassium. - Plant mol. Biol. 87: 317-327, 2015. Go to original source...
    44. Zhou, S., Zhang, Z., Tang, Q., Lan, H., Li, Y., Luo, P.: Enhanced V-ATPase activity contributes to the improved salt tolerance of transgenic tobacco plants overexpressing vacuolar Na+/H+ antiporter AtNHX1. - Biotechnol. Lett. 33: 375-380, 2011. Go to original source...
    45. Zhu, J.K.: Regulation of ion homeostasis under salt stress. - Curr. Opin. Plant Biol. 6: 441-445, 2003. Go to original source...

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