Journal of Plant Biology

, Volume 51, Issue 2, pp 159–165 | Cite as

Overexpression ofENA1 from yeast increases salt tolerance inArabidopsis

  • Xiangqiang Kong
  • Xiuhua Gao
  • Weihuan Li
  • Jiqiang Zhao
  • Yanxiu Zhao
  • Hui Zhang


In yeast, the plasma membrane Na+/H+ antiporter and Na+-ATPase are key enzymes for salt tolerance.Saccharomyces cerevisiae Na+-ATPase (Enalp ATPase) is encoded by theENA1/PMR2A gene; expression ofENA1 is tightly regulated by Na+ and depends on ambient pH. Although Enalp is active mainly at alkaline pH values inS. cerevisiae, no Na+-ATPase has been found in flowering plants. To test whether this yeast enzyme would improve salt tolerance in plants, we introducedENA1 intoArabidopsis (cv. Columbia) under the control of the cauliflower mosaic virus 35S promoter. Transformants were selected for their ability to grow on a medium containing kanamyin. Southern blot analyses confirmed thatENA1 was transferred into theArabidopsis genome and northern blot analyses showed thatENA1 was expressed in the transformants. Several transgenic homozygous lines and wild-type (WT) plants were evaluated for salt tolerance. No obvious morphological or developmental differences existed between the transgenic and WT plants in the absence of stress. However, overexpression ofENA1 inArabidopsis improved seed germination rates and salt tolerance in seedlings. Under saline conditions, transgenic plants accumulated a lower amount of Na+ than did the wild type, and fresh and dry weights of the former were higher. Other experiments revealed that expression ofENA1 promoted salt tolerance in transgenicArabidopsis under both acidic and alkaline conditions.


alkaline pH Arabidopsis Na+-ATPase Saccharomyces cerevisiae salt tolerance 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Literature Cited

  1. Apse MP, Aharon GS, Snedden WA, Blumwald E (1999) Salt tolerance conferred by overexpression of a vacuolar Na+/H+ anti-porter inArabidopsis. Science285: 1256–1258PubMedCrossRefGoogle Scholar
  2. Banuelos MA, Rodrfguez-Navarro A (1998) P-type ATPase mediate sodium and potassium effluxes inSaccharomyces cerevisiae. J Biol Chem273: 1640–1646PubMedCrossRefGoogle Scholar
  3. Banuelos MA, Quintero FJ, Rodríguez-Navarro A (1995) Functional expression of theENA1 (PMR2)-ATPase ofSaccharomyces cerevisiae inSchizosaccharomyces pombe. Biochim Biophys Acta1229: 233–238PubMedCrossRefGoogle Scholar
  4. Berthomieu P;Conejero G, Nublat A, Brackenbury WJ, Lambert C, Savio C, Uozomi N, Oiki S, Yamada K, Cellier F (2003) Functional analysis ofAtHKT1 inArabidopsis shows that Na+ recirculation by the phloem is crucial for salt tolerance. EMBO J22: 2004–2014PubMedCrossRefGoogle Scholar
  5. Chomczynski P, Sacci N (1987) Single-step method of RNA isolation by acid guanidiumthiocyanate-phenol-chloroform extraction. Anal Biochem162: 156–159PubMedCrossRefGoogle Scholar
  6. Clough SJ, Bent AF (1998) Floral dip: A simplified method forAgrobacterium-mediated transformation ofArabidopsis thaliana. Plant J16: 735–743PubMedCrossRefGoogle Scholar
  7. Flowers TJ, Yeo AR (1995) Breeding for salinity resistance in crop plants: Where next? Aust J Plant Physiol22: 875–884CrossRefGoogle Scholar
  8. Flowers TJ, Troke PF, Yeo AR (1977) The mechanism of salt tolerance in halophytes. Annu Rev Plant Physiol22: 89–121CrossRefGoogle Scholar
  9. Fukuda A, Nakamura A, Tanaka Y (1999) Molecular cloning and expression of the Na+/H+ exchanger gene inOryza sativa. Biochim Biophys Acta 1446: 149–155PubMedGoogle Scholar
  10. Gao XH, Ren ZH, Zhao YX, Zhang H (2003) Overexpression ofSOD2 increase salt toleranceof Arabidopsis. Plant Physiol133: 1873–1881PubMedCrossRefGoogle Scholar
  11. Guo Y, Qiu Q, Quintero FJ, Pardo JM, Ohta M, Zhang C, Schumaker KS, Zhu JK (2004) Transgenic evaluation of activated mutant alleles of SOS2 reveals a critical requirement for its kinase activity and C-terminal regulatory domain for salt tolerance inArabidopsis thaliana. Plant Cell16: 435–449PubMedCrossRefGoogle Scholar
  12. Hamada A, Shono M, Xia T, Ohta M, Hayashi Y, Tanaka A, Hayakawa T (2001) Isolation and characterization of Na+/H+ anti-porter gene from the halophyteAtriplex gmelini. Plant Mol Biol46: 35–42PubMedCrossRefGoogle Scholar
  13. Haro R, Garcia de Blas B, Rodríguez-Navarro A (1991) A novel P- type ATPase from yeast involved in sodium transport. FEBS Lett291: 189–191PubMedCrossRefGoogle Scholar
  14. Hasegawa PM, Bressan RA, Zhu JK, Bohnert HJ (2000) Plant cellular and molecular responses to high salinity. Annu Rev Plant Physiol Plant Mol Biol51: 463–499PubMedCrossRefGoogle Scholar
  15. Jia ZP, Mccullough N, Martel R, Hemmingsen S, Young PG (1992) Gene amplification at a locus encoding a putative Na+/H+ antiporter confers sodium and lithium tolerance in fission yeast. EMBO J11: 1631–1640PubMedGoogle Scholar
  16. Katiyar-Agarwal S, Zhu JH, Kim K, Agarwal M, Fu XM, Huang A, Zhu JK (2006) The plasma membrane Na+/H+ antiporter SOS1 interacts with RCD1 and functions in oxidative stress tolerance inArabidopsis. Proc Natl Acad Sci USA103: 18816–18821PubMedCrossRefGoogle Scholar
  17. Ma XL, Zhang Q, Shi HZ, Zhu JK, Zhao YX, Ma CL, Zhang H (2004) Molecular cloning and different expression of a vacuolar Na+/H+ antiporter gene inSuaeda salsa under salt stress. Biol Plant48: 219–225CrossRefGoogle Scholar
  18. Munns R (2002) Comparative physiology of salt and water stress. Plant Cell Environ25: 239–250PubMedCrossRefGoogle Scholar
  19. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol Plant15: 473–497CrossRefGoogle Scholar
  20. Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant cDNA. Nucleic Acids Res8: 4321–4325PubMedCrossRefGoogle Scholar
  21. Nakayama H, Yoshida K, Shinmyo A (2004) Yeast plasma membrane Ena1p ATPase alters alkali-cation homeostasis and confers increased salt tolerance in tobacco cultured cells. Biotechnol Bioengr85: 776–789CrossRefGoogle Scholar
  22. Qiu Q, Guo Y, Dietrich MA, Schumaker KS, Zhu JK (2002) Regulation of SOS1, a plasma membrane Na+/H+ exchanger inArabidopsis thaliana, by SOS2 and SOS3. Proc Natl Acad Sci USA99: 8436–8441PubMedCrossRefGoogle Scholar
  23. Ramos J (1999) Contrasting salt tolerance mechanisms inSaccharomyces cerevisiae andDebaryomyces hansenii. Recent Res Dev Microbiol3: 377–390Google Scholar
  24. Ren ZH, Gao JP, Li LG, Cai XL, Huang W, Chao DY, Zhu MZ, Wang ZY, Luan S, Lin HX (2005) A rice quantitative trait locus for salt tolerance encodes a sodium transporter. Nat Genet37: 1141–1146PubMedCrossRefGoogle Scholar
  25. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: A Laboratory Manual, Ed 2. Cold Spring Harbor Laboratory Press, New YorkGoogle Scholar
  26. Serrano R (1996) Salt tolerance in plants and microorganisms: Toxicity targets and defense responses. Intl Rev Cyto165: 1–52CrossRefGoogle Scholar
  27. Serrano R, Rodrfguez-Navarro A (2001) Ion homeostasis during salt stress in plants. Curr Opin Cell Biol13: 399–404PubMedCrossRefGoogle Scholar
  28. Shi H, Wu SJ, Zhu JK (2003) Overexpression of a plasma membrane Na+/H+ antiporter improves salt tolerance inArabidopsis. Nat Biotechnol21: 81–85PubMedCrossRefGoogle Scholar
  29. Troll W, Lindsley J (1995) Proline content determination in plant tissues. J Biol Chem215: 655–660Google Scholar
  30. Wang BS, Zhao KF (1995) Comparison of extractive methods of Na+, K+ in wheat leaves. Plant Physiol Commun31: 50–52Google Scholar
  31. Zhang HX, Blumwald E (2001) Transgenic salt-tolerance tomato plants accumulate salt in foliage but not in fruit. Nat Biotechnol19: 765–768PubMedCrossRefGoogle Scholar
  32. Zhang HX, Hodson JN, Williams JP, Blumwald E (2001) Engineering salt-tolerantBrassica plants: Characterization of yield and seed oil quality in transgenic plants with increased vacuolar sodium accumulation. Proc Natl Acad Sci USA98: 12832–12836PubMedCrossRefGoogle Scholar
  33. Zhu JK (2000) Genetic analysis of plant salt tolerance usingArabidopsis. Plant Physiol124: 941–948PubMedCrossRefGoogle Scholar
  34. Zhu JK (2001) Plant salt tolerance. Trends Plant Sci6: 66–71PubMedCrossRefGoogle Scholar
  35. Zhu JK (2002) Salt and drought stress signal transduction in plants. Annu Rev Plant Biol53: 247–273PubMedCrossRefGoogle Scholar

Copyright information

© The Botanical Society of Korea 2008

Authors and Affiliations

  • Xiangqiang Kong
    • 1
  • Xiuhua Gao
    • 1
  • Weihuan Li
    • 1
  • Jiqiang Zhao
    • 1
  • Yanxiu Zhao
    • 1
  • Hui Zhang
    • 2
    • 1
  1. 1.Kay Lab of Plant Stress ResearchCollege of Life Science, Shandong Normal UniversityShandong ProvincePR China
  2. 2.Life Science College of Shandong Normal UniversityShandong ProvincePR China

Personalised recommendations