Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

The plasma membrane–bound phospholipase Dδ enhances freezing tolerance in Arabidopsis thaliana

Nature Biotechnology volume 22, pages 427–433 (2004)Cite this article

Abstract

Freezing injury is a major environmental limitation on the productivity and geographical distribution of plants. Here we show that freezing tolerance can be manipulated in Arabidopsis thaliana by genetic alteration of the gene encoding phospholipase Dδ (PLDδ), which is involved in membrane lipid hydrolysis and cell signaling. Genetic knockout of the plasma membrane–associated PLDδ rendered A. thaliana plants more sensitive to freezing, whereas overexpression of PLDδ increased freezing tolerance. Lipid profiling revealed that PLDδ contributed approximately 20% of the phosphatidic acid produced in wild-type plants during freezing, and overexpression of PLDδ increased the production of phosphatidic acid species. The PLDδ alterations did not affect the expression of the cold-regulated genes COR47 or COR78 or alter cold-induced increases in proline or soluble sugars, suggesting that the PLD pathway is a unique determinant of the response to freezing and may present opportunities for improving plant freezing tolerance.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Decreased freezing tolerance in PLDδ-KO A. thaliana.
Figure 2: Increased freezing sensitivity of cold-acclimated PLDδ-KO seedlings of A. thaliana.
Figure 3: Overexpression of PLDδ (PLDδ-OE) and the effect of cold acclimation and freezing on the PLDδ expression in A. thaliana.
Figure 4: Increased freezing tolerance in PLDδ-overexpressing plants (PLDδ-OE).
Figure 5: Effect of PLDδ-KO and PLDδ-OE on the expression of cold-regulated genes and H2O2-induced cell death.
Figure 6: Changes in phosphatidic acid (PA) species and proline and soluble sugar content after cold treatments of PLDδ-KO, PLDδ-OE and wild-type A. thaliana.

Similar content being viewed by others

References

  1. Thomashow, M.F. Plant cold acclimation: freezing tolerance genes and regulatory mechanisms. Annu. Rev. Plant Physiol. Plant Mol. Biol. 50, 571–599 (1999).

    Article  CAS  Google Scholar 

  2. Iba, K. Acclimative response to temperature stress in higher plants: approaches of gene engineering for temperature tolerance. Annu. Rev. Plant Biol. 53, 225–245 (2002).

    Article  CAS  Google Scholar 

  3. Gong, Z. et al. RNA helicase-like protein as an early regulator of transcription factors for plant chilling and freezing tolerance. Proc. Natl. Acad. Sci. USA 99, 11507–11512 (2002).

    Article  CAS  Google Scholar 

  4. Chinnusamy, V. et al. ICE1: a regulator of cold-induced transcriptome and freezing tolerance in Arabidopsis. Genes Dev. 17, 1043–1054 (2003).

    Article  CAS  Google Scholar 

  5. Jaglo-Ottosen, K.R., Gilmour, S.J., Zarka, D.G., Schabenberger, O. & Thomashow, M.F. Arabidopsis CBF1 overexpression induces COR genes and enhances freezing tolerance. Science 280, 104–106 (1998).

    Article  CAS  Google Scholar 

  6. Kasuga, M., Liu, Q., Miura, S., Yamaguchi-Shinozaki, K. & Shinozaki, K. Improving plant drought, salt, and freezing tolerance by gene transfer of a single stress-inducible transcription factor. Nat. Biotechnol. 17, 287–291 (1999).

    Article  CAS  Google Scholar 

  7. Xin, Z. & Browse, J. eskimo1 mutants of Arabidopsis are constitutively freezing-tolerant. Proc. Natl. Acad. Sci. USA 95, 7799–7804 (1998).

    Article  CAS  Google Scholar 

  8. Seki, M. et al. Monitoring the expression pattern of 1300 Arabidopsis genes under drought and cold stresses by using a full-length cDNA microarray. Plant Cell 13, 61–72 (2001).

    Article  CAS  Google Scholar 

  9. Fowler, S. & Thomashow, M.F. Arabidopsis transcriptome profiling indicates that multiple regulatory pathways are activated during cold acclimation in addition to the CBF cold response pathway. Plant Cell 14, 1675–1690 (2002).

    Article  CAS  Google Scholar 

  10. Thomashow, M.F. So what's new in the field of plant cold acclimation? Lots! Plant Physiol. 125, 89–93 (2001).

    Article  CAS  Google Scholar 

  11. Knight. H., Trewavas, A.J. & Knight, M. Cold calcium signaling in Arabidopsis involves two cellular pools and a change in calcium signature after acclimation. Plant Cell 8, 489–503 (1996).

    CAS  PubMed  PubMed Central  Google Scholar 

  12. Plieth, C., Hansen, U.P., Knight, H. & Knight, M. Temperature sensing by plants: the primary characteristics of signal perception and calcium response. Plant J. 18, 491–497 (1999).

    Article  CAS  Google Scholar 

  13. Ruelland, E., Cantrel, C., Gawer, M., Kader, J.C. & Zachowski, A. Activation of phospholipases C and D is an early response to a cold exposure in Arabidopsis suspension cells. Plant Physiol. 130, 999–1007 (2002).

    Article  CAS  Google Scholar 

  14. Welti, R. et al. Profiling membrane lipids in plant stress responses. Role of phospholipase Dα in freezing-induced lipid changes in Arabidopsis. J. Biol. Chem. 277, 31994–32002 (2002).

    Article  CAS  Google Scholar 

  15. Cullis, P.R. & DeKruijff, B. Lipid polymorphism and the functional roles of lipids in biological membranes. Biochim. Biophys. Acta 559, 399–420 (1979).

    Article  CAS  Google Scholar 

  16. Verleij, A.J., DeMaagd, R., Leunissen-Bijvelt, J. & DeKruijff, B. Divalent cations and chlorpromazine can induce non-bilayer structures in phosphatidic acid-containing membranes. Biochim. Biophys. Acta 684, 255–262 (1982).

    Article  Google Scholar 

  17. Wang, X. Phospholipase D in hormonal and stress signaling. Curr. Opin. Plant Biol. 5, 408–414 (2002).

    Article  CAS  Google Scholar 

  18. Qin, C. & Wang, X. The Arabidopsis phospholipase D family. Characterization of a calcium-independent and phosphatidylcholine-selective PLDζ1 with distinct regulatory domains. Plant Physiol. 128, 1057–1068 (2002).

    Article  CAS  Google Scholar 

  19. Wang, C. & Wang, X. A novel phospholipase D of Arabidopsis that is activated by oleic acid and associated with the plasma membrane. Plant Physiol. 127, 1102–1112 (2001).

    Article  CAS  Google Scholar 

  20. Gardiner, J.C. et al. 90-kD phospholipase D from tobacco binds to microtubules and the plasma membrane. Plant Cell 13, 2143–2158 (2001).

    Article  CAS  Google Scholar 

  21. Katagiri, T., Takahashi, S. & Shinozaki, K. Involvement of a novel Arabidopsis phospholipases D, AtPLD, in dehydration-inducible accumulation of phosphatidic acid in stress signaling. Plant J. 26, 595–605 (2001).

    Article  CAS  Google Scholar 

  22. Zhang, W. et al. Phospholipase Dδ and phosphatidic acid decrease H2O2-induced cell death in Arabidopsis. Plant Cell 15, 2285–2295 (2003).

    Article  CAS  Google Scholar 

  23. Prasad, T.K., Anderson, M.D., Martin, B.A. & Stewart, C.R. Evidence for chilling-induced oxidative stress in maize seedlings and a regulatory role for hydrogen peroxide. Plant Cell 6, 65–74 (1994).

    Article  CAS  Google Scholar 

  24. Sakamoto, A., Valverde, R., Alia, R., Chen, T.H.H. & Murata, N. Transformation of Arabidopsis with the codA gene for choline oxidase enhances freezing tolerance of plants. Plant J. 22, 449–453 (2000).

    Article  CAS  Google Scholar 

  25. McKersie, B.D., Bowley, S.R. & Jones, K.S. Winter survival of transgenic alfalfa overexpressing superoxide dismutase. Plant Physiol. 119, 839–848 (1999).

    Article  CAS  Google Scholar 

  26. Dubouzet, J.G. et al. OsDREB genes in rice, Oryza sativa L., encode transcription activators that function in drought-, high-salt- and cold-responsive gene expression. Plant J. 33, 751–763 (2003).

    Article  CAS  Google Scholar 

  27. Hardy, S., Langelier, Y. & Prentki, M. Oleate activates phosphatidylinositol 3-kinase and promotes proliferation and reduces apoptosis of MDA-MB-231 breast cancer cells, whereas palmitate has opposite effects. Cancer Res. 60, 6353–6358 (2000).

    CAS  PubMed  Google Scholar 

  28. Kachroo, P., Shanklin, J., Shah, J., Whittle, E.J. & Klessig, D.F. A fatty acid desaturase modulates the activation of defense signaling pathways in plants. Proc. Natl. Acad. Sci. USA 98, 9448–9453 (2001).

    Article  CAS  Google Scholar 

  29. Fan, L., Zheng, S. & Wang, X. Antisense suppression of phospholipase Dα retards abscisic acid- and ethylene- promoted senescence in postharvest Arabidopsis leaves. Plant Cell 9, 2183–2196 (1997).

    CAS  PubMed  PubMed Central  Google Scholar 

  30. Pappan, K., Zheng, S. & Wang, X. Identification and characterization of a novel phospholipase D that requires polyphosphoinositides and submicromolar calcium for activity in Arabidopsis. J. Biol. Chem. 272, 7048–7054 (1997).

    Article  CAS  Google Scholar 

  31. Magne, C. & Larher, F. High sugar content of extracts interferes with colorimetric determination of amino acids and free proline. Anal. Biochem. 200, 115–118 (1992).

    Article  CAS  Google Scholar 

  32. Bravo, L.A., Zúñiga, G.E., Alberdi, M. & Corcuera, L.J. The role of ABA in freezing tolerance and cold acclimation in barley. Physiol. Plant. 103, 17–23 (1998).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from the National Science Foundation (NSF), US Department of Agriculture, Kansas NSF EPSCoR and the Kansas State University Plant Biotechnology Center. The authors would like to thank Charles Rife for freezing chamber use, Todd Williams for acquisition of the ESI-MS/MS data and Christen Buseman for help with processing of the lipid profiling data. This is contribution 04-265-J from the Kansas Agricultural Experiment Station.

Author information

Authors and Affiliations

  1. Department of Biochemistry, Kansas State University, Manhattan, 66506-3702, Kansas, USA

    Weiqi Li, Maoyin Li, Wenhua Zhang & Xuemin Wang

  2. Division of Biology, Kansas State University, Manhattan, 66506-3702, Kansas, USA

    Ruth Welti

Authors
  1. Weiqi Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Maoyin Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wenhua Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ruth Welti
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Xuemin Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Xuemin Wang.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Li, W., Li, M., Zhang, W. et al. The plasma membrane–bound phospholipase Dδ enhances freezing tolerance in Arabidopsis thaliana. Nat Biotechnol 22, 427–433 (2004). https://doi.org/10.1038/nbt949

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt949

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing