Skip to main content

Advertisement

Springer Nature Link
Log in

Improved Growth, Drought Tolerance, and Ultrastructural Evidence of Increased Turgidity in Tobacco Plants Overexpressing Arabidopsis Vacuolar Pyrophosphatase (AVP1)

  • Research
  • Published:
Molecular Biotechnology Aims and scope Submit manuscript

Abstract

An increasing volume of evidence indicating the mechanisms of drought tolerance of AVP1-overexpressing transgenic plants has been reported. In the present study, we are reporting the experiments conducted for the drought tolerance of AVP1 overexpressing plants and WT tobacco plants in three water regimes named as “fully watered,” “less-watered,” and “desiccated”. Results suggest that AVP1 plants exhibited greater vigor and drought tolerance in quantitative terms i.e., increase in size and weight of shoots and capsules. AVP1 plants produced more seeds than WT across all three water regimes. The less-watered regime was found to produce the greatest contrast. AVP1 overexpression enhanced solute accumulation in vacuoles resulting in an increase in water retention and turgor of the cell. The ultrastructure study of AVP1 overexpressing cells and WT leaf cells revealed that AVP1 plants displayed more turgid and hyperosmotic cells than WT. Moreover, guard cells in the AVP1 plants exhibited thick cell walls, few vacuoles, and deep and close stomata, whereas WT plants showed larger vacuoles and relatively open stomata aperture with no significant difference in size and number of the cells per unit area.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+
from $39.99 /Month
  • Starting from 10 chapters or articles per month
  • Access and download chapters and articles from more than 300k books and 2,500 journals
  • Cancel anytime
View plans

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10

Similar content being viewed by others

Explore related subjects

Discover the latest articles, books and news in related subjects, suggested using machine learning.

References

  1. Levitt, J. (1980). Responses of plants to environmental stresses (2nd ed.). New York: Academic Press.

    Google Scholar 

  2. Alpert, P. (2000). The discovery, scope and puzzle of desiccation tolerance in plants. Plant Ecology, 151, 5–17.

    Article  Google Scholar 

  3. Hsiao, T. C. (1973). Plant responses to water stress. Annual Review of Plant Physiology, 24, 519–570.

    Article  CAS  Google Scholar 

  4. Schulze, E. D. (1986). Carbon dioxide and water vapor exchange in response to drought in the atmosphere and in the soil. Annual Review of Plant Physiology, 37, 247–274.

    Article  Google Scholar 

  5. Cosgrove, D. J. (2000). Loosening of plant cell walls by expansins. Nature, 407, 321–326.

    Article  CAS  Google Scholar 

  6. Vicre, M., Sherwin, H. W., Driouich, A., Jaffer, M., Jauneau, A., & Farrant, J. M. (1999). Cell wall properties of hydrated and dry leaves of the resurrection plant Craterostigma wilmsii. Journal of Plant Physiology, 155, 719–726.

    Article  CAS  Google Scholar 

  7. Sarafian, V., Kim, Y., Poole, R. J., & Rea, P. A. (1992). Molecular cloning and sequence of cDNA encoding the pyrophosphate-energized vacuolar membrane proton pump of Arabidopsis thaliana. Proceedings National Academy of Sciences, 89, 1775–1779.

    Article  CAS  Google Scholar 

  8. Vercruyssen, L., Gonzalez, N., Werner, T., Schmülling, T., & Inzé, D. (2011). Combining enhanced root and shoot growth reveals cross talk between pathways that control plant organ size in Arabidopsis. Plant Physiology, 155(3), 1339–1352.

    Article  CAS  Google Scholar 

  9. Dong, Q.-L., Liu, D.-D., An, X.-H., Hu, D.-G., Yao, Y.-X., & Hao, Y.-J. (2011). MdVHP1 encodes an apple vacuolar H+-PPase and enhances stress tolerance in transgenic apple callus and tomato. Journal of Plant Physiology, 168(17), 2124–2133.

    Article  CAS  Google Scholar 

  10. Yao, Y. X., Dong, Q. L., You, C. X., Zhai, H., & Hao, Y. J. (2011). Expression analysis and functional characterization of apple MdVHP1 gene reveals its involvement in Na(+), malate and soluble sugar accumulation. Plant Physiology and Biochemistry, 49(10), 1201–1208.

    Article  CAS  Google Scholar 

  11. Liu, L., Wang, Y., Wang, N., Dong, Y. Y., Fan, X. D., Liu, X. M., et al. (2011). Cloning of a vacuolar H(+)-pyrophosphatase gene from the halophyte Suaeda corniculata whose heterologous overexpression improves salt, saline-alkali and drought tolerance in Arabidopsis. Journal of Integrated Plant Biology, 53(9), 731–742.

    CAS  Google Scholar 

  12. Zhang, J., Li, J., Wang, X., & Chen, J. (2011). OVP1, a vacuolar H+-translocating inorganic pyrophosphatase (V-PPase) overexpression improved rice cold tolerance. Plant Physiology and Biochemistry, 49(1), 33–38.

    Article  Google Scholar 

  13. Pasapula, V., Shen, G., Kuppu, S., Paez-Valencia, J., Mendoza, M., Hou, P., et al. (2011). Expression of an Arabidopsis vacuolar H+-pyrophosphatase gene (AVP1) in cotton improves drought and salt tolerance and increases fibre yield in the field conditions. Plant Biotechnology Journal, 9(1), 88–99.

    Article  CAS  Google Scholar 

  14. Li, Z., Baldwin, C. M., Hu, Q., Liu, H., & Luo, H. (2010). Heterologous expression of Arabidopsis H+-pyrophosphatase enhances salt tolerance in transgenic creeping bentgrass (Agrostis stolonifera L.). Plant Cell Environment, 33(2), 272–289.

    Article  CAS  Google Scholar 

  15. Lv, S. L., Lian, L. J., Tao, P. L., Li, Z. X., Zhang, K. W., & Zhang, J. R. (2009). Overexpression of Thellungiella halophila H(+)-PPase (TsVP) in cotton enhances drought stress resistance of plants. Planta, 229(4), 899–910.

    Article  CAS  Google Scholar 

  16. Lv, S., Zhang, K., Gao, Q., Lian, L., Song, Y., & Zhang, J. (2008). Overexpression of an H+-PPase gene from Thellungiella halophila in cotton enhances salt tolerance and improves growth and photosynthetic performance. Plant Cell Physiology, 49(8), 1150–1164.

    Article  CAS  Google Scholar 

  17. Brini, F., Hanin, M., Mezghani, I., Berkowitz, G. A., & Masmoudi, K. (2007). Overexpression of wheat Na+/H+ antiporter TNHX1 and H+-pyrophosphatase TVP1 improve salt- and drought-stress tolerance in Arabidopsis thaliana plants. Journal of Experimental Botany, 58(2), 301–308.

    Article  CAS  Google Scholar 

  18. Gao, F., Gao, Q., Duan, X., Yue, G., Yang, A., & Zhang, J. (2006). Cloning of an H+-PPase gene from Thellungiella halophila and its heterologous expression to improve tobacco salt tolerance. Journal of Experimental Botany, 57(12), 3259–3270.

    Article  CAS  Google Scholar 

  19. Guo, S., Yin, H., Zhang, X., Zhao, F., Li, P., Chen, S., et al. (2006). Molecular cloning and characterization of a vacuolar H+-pyrophosphatase gene, SsVP, from the halophyte Suaeda salsa and its overexpression increases salt and drought tolerance of Arabidopsis. Plant Molecular Biology, 60(1), 41–50.

    Article  CAS  Google Scholar 

  20. Park, S., Li, J., Pittman, J. K., Berkowitz, G. A., Yang, H., Undurraga, S., et al. (2005). Up-regulation of a H+-pyrophosphatase (H+-PPase) as a strategy to engineer drought-resistant crop plants. Proceedings National Academy of Sciences, 102(52), 18830–18835.

    Article  CAS  Google Scholar 

  21. Gaxiola, R. A., Li, J., Undurraga, S., Dang, L. M., Allen, G. J., Alper, S. L., et al. (2001). Drought and salt-tolerant plants result from overexpression of the AVP1 H+-pump. Proceedings National Academy of Sciences, 98, 11444–11449.

    Article  CAS  Google Scholar 

  22. Voelker, T., Sturm, A., & Chrispeels, M. J. (1987). Differences in expression between two seed lectin alleles obtained from normal and lectin-deficient beans are maintained in transgenic tobacco. EMBO Journal, 6, 3571–3577.

    CAS  Google Scholar 

  23. Kunik, T., Salomon, R., Zamir, D., Navot, N., Zeidan, M., Michelson, I., et al. (1994). Transgenic tomato plants expressing the tomato yellow leaf curl virus capsid protein are resistant to the virus. Bio-Technology, 12, 500–504.

    Article  CAS  Google Scholar 

  24. Verwoerd, T., Dekker, B. M. M., & Hoekema, A. (1989). A small-scale procedure for the rapid isolation of plant RNAs. Nucleic Acid Research, 17, 2362.

    Article  CAS  Google Scholar 

  25. Bradford, M. M. (1976). A rapid and sensitive method for the quantification of microgram quantities of protein, utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248–254.

    Article  CAS  Google Scholar 

  26. Maeshima, M. (2000). Vacuolar H+-pyrophosphatase. Biochimica et Biophysica Acta, 1465, 37–51.

    Article  CAS  Google Scholar 

  27. Spurr, A. R. (1969). A low viscosity epoxy resin embedding medium for electron microscopy. Journal of Ultrastructure Research, 26, 31–43.

    Article  CAS  Google Scholar 

  28. Roberts, I. M. (2002). Iso-butanol saturated water: A simple procedure for increasing staining intensity of resin sections for light and electron microscopy. Journal of Microscopy, 207, 97–107.

    Article  CAS  Google Scholar 

  29. Hameed, S. (2003). Molecular characterization and significance of rhizobial exopolysaccharides. PhD thesis, Quaid-i-Azam University, Islamabad, pp. 28–30.

  30. Sievers, F., Wilm, A., Dineen, D. G., Gibson, T. J., Karplus, K., Li, W., et al. (2011). Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega. Molecular Systems Biology, 7, 539.

    Article  Google Scholar 

  31. Li, F., Xing, S., Guo, Q., Zhao, M., Zhang, J., Gao, Q., et al. (2011). Drought tolerance through over-expression of the expansin gene TaEXPB23 in transgenic tobacco. Journal of Plant Physiology, 168(9), 960–966.

    Article  CAS  Google Scholar 

  32. Lucas, S., Durmaz, E., Akpinar, B. A., & Budak, H. (2011). The drought response displayed by a DRE-binding protein from Triticum dicoccoides. Plant Physiology Biochemistry, 49, 346–351.

    Article  CAS  Google Scholar 

  33. Rivero, R. M., Kojima, M., Gepstein, A., Sakakibara, H., Mittler, R., Gepstein, S., et al. (2007). Delayed leaf senescence induces extreme drought tolerance in a flowering plant. Proceedings of the National Academy of Sciences, 104, 19631–19636.

    Article  CAS  Google Scholar 

  34. Zhu, X. G., Long, S. P., & Ort, D. R. (2010). Improving photosynthetic efficiency for greater yield. Annual Review of Plant Biology, 61, 235–261.

    Article  CAS  Google Scholar 

  35. Li, J., Yang, H., Peer, W. A., Richter, G., Blakeslee, J., Bandyopadhyay, A., et al. (2005). Arabidopsis H+-PPase AVP1 regulates auxin-mediated organ development. Science, 310(5745), 121–125.

    Article  CAS  Google Scholar 

  36. Lewis, J. K., & Valerie, M. K. (1995). The cell surface and cellular communication. In: Principles and molecular biology (2nd ed., pp. 242–245). New York: Benjamin-Cummings.

  37. Couot-Gastelier, J., Laffray, D., & Louguet, P. (1984). Etude comparee de 1′utrastructure des stomates ouverts et fermes chez le Tradescantia viragimiana. Canadian Journal of Botany, 62(1505–1512), 41.

    Google Scholar 

  38. Meidner, H., & Edwards, M. (1975). Direct measurements of turgor pressure potentials of guard cells, I. Journal of Experimental Botany, 26, 319–330.

    Article  Google Scholar 

  39. Edwards, M., Meidner, H., & Sheriff, D. W. (1976). Direct measurement of turgor pressure potentials of guard cells: II. The mechanical advantage of subsidiary cells, the spannungsphase, and the optimum leaf water deficit. Journal of Experimental Botany, 27, 163–171.

    Article  Google Scholar 

  40. Franks, P. J., Cowan, I. R., Tyerman, S. D., Cleary, A. L., Lloyd, J., & Farquhar, G. D. (1995). Guard cell pressure/aperture characteristics measured with the pressure probe. Plant Cell Environment, 18, 795–800.

    Article  Google Scholar 

  41. Franks, P. J., Cowanl, R. I., & Farquhar, G. D. (1998). A study of stomatal mechanics using the cell pressure probe. Plant, Cell and Environment, 21, 94–100.

    Article  Google Scholar 

  42. Gao, Xin.-Qi., Li, Chun.-Guang., Wei, Peng.-Cheng., Zhang, Xin.-Yan., Chen, Jia., Wang, Xue.-Chen., et al. (2005). The dynamic changes of tonoplasts in guard cells are important for stomatal movement in Vicia faba. Plant Physiology, 139(3), 1207–1216.

    Article  CAS  Google Scholar 

  43. Gaxiola, R. A., Fink, G. R., & Hirschi, K. D. (2002). Genetic manipulation of vacuolar proton pumps and transporters. Plant Physiology, 129, 967–973.

    Article  CAS  Google Scholar 

  44. Blatt, M. R. (2000). Cellular signaling and volume control in stomatal movements in plants. Annual Review of Cell Developmental Biology, 16, 221–241.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors acknowledge Dr. Roberto Gaxiola, Assistant Professor Cellular and Molecular Biosciences School of Life Sciences, Arizona State University, Arizona, USA for providing us with AVP1 gene construct. The authors also acknowledge Dr. Sohail Hameed Incharge, Electron Microscopy services and Mr. Javed Iqbal at National Institute for Biotechnology and Genetic Engineering (NIBGE), Pakistan for their technical support in electron microscopy experiments. We thank Pak–US project for financial support of this work in the laboratory of Prof. Dr. Eduardo Blumwald, Department of Plant Sciences, University of California, Davis USA.

Author information

Authors and Affiliations

  1. National Institute for Biotechnology and Genetic Engineering, Faisalabad, Pakistan

    Anjuman Arif & Muhammad Arif

  2. Biosciences Division, Pakistan Atomic Energy Commission (PAEC), Islamabad, Pakistan

    Yusuf Zafar

  3. Department of Plant Sciences, University of California, Davis, CA, 95616, USA

    Eduardo Blumwald

Authors
  1. Anjuman Arif
    View author publications

    Search author on:PubMed Google Scholar

  2. Yusuf Zafar
    View author publications

    Search author on:PubMed Google Scholar

  3. Muhammad Arif
    View author publications

    Search author on:PubMed Google Scholar

  4. Eduardo Blumwald
    View author publications

    Search author on:PubMed Google Scholar

Corresponding author

Correspondence to Anjuman Arif.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Arif, A., Zafar, Y., Arif, M. et al. Improved Growth, Drought Tolerance, and Ultrastructural Evidence of Increased Turgidity in Tobacco Plants Overexpressing Arabidopsis Vacuolar Pyrophosphatase (AVP1). Mol Biotechnol 54, 379–392 (2013). https://doi.org/10.1007/s12033-012-9577-9

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12033-012-9577-9

Keywords