Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Phospholipases Dζ1 and Dζ2 have distinct roles in growth and antioxidant systems in Arabidopsis thaliana responding to salt stress

Abstract

Main conclusion

Phospholipases Dζ play different roles in Arabidopsis salt tolerance affecting the regulation of ion transport and antioxidant responses.

Lipid signalling mediated by phospholipase D (PLD) plays essential roles in plant growth including stress and hormonal responses. Here we show that PLDζ1 and PLDζ2 have distinct effects on Arabidopsis responses to salinity. A transcriptome analysis of a double pldζ1pldζ2 mutant revealed a cluster of genes involved in abiotic and biotic stresses, such as the high salt-stress responsive genes DDF1 and RD29A. Another cluster of genes with a common expression pattern included ROS detoxification genes involved in electron transport and biotic and abiotic stress responses. Total superoxide dismutase (SOD) activity was induced early in the shoots and roots of all pldζ mutants exposed to mild or severe salinity with the highest SOD activity measured in pldζ2 at 14 days. Lipid peroxidation in shoots and roots was higher in the pldζ1 mutant upon salt treatment and pldζ1 accumulated H2O2 earlier than other genotypes in response to salt. Salinity caused less deleterious effects on K+ accumulation in shoots and roots of the pldζ2 mutant than of wild type, causing only a slight variation in Na+/K+ ratio. Relative growth rates of wild-type plants, pldζ1, pldζ2 and pldζ1pldζ2 mutants were similar in control conditions, but strongly affected by salt in WT and pldζ1. The efficiency of photosystem II, estimated by measuring the ratio of chlorophyll fluorescence (F v/F m ratio), was strongly decreased in pldζ1 under salt stress. In conclusion, PLDζ2 plays a key role in determining Arabidopsis sensitivity to salt stress allowing ion transport and antioxidant responses to be finely regulated.

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

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

Abbreviations

MDA:

Malondialdehyde

PA:

Phosphatidic acid

PLD:

Phospholipase D

SOD:

Superoxide dismutase

References

  1. Alonso JM, Stepanova AN (2003) T-DNA mutagenesis in Arabidopsis. Methods Mol Biol Clifton NJ 236:177–188. doi:10.1385/1-59259-413-1:177

  2. Anschütz U, Becker D, Shabala S (2014) Going beyond nutrition: regulation of potassium homoeostasis as a common denominator of plant adaptive responses to environment. J Plant Physiol 171:670–687

  3. Bargmann BO, Munnik T (2006) The role of phospholipase D in plant stress responses. Curr Opin Plant Biol 9:515–522. doi:10.1016/j.pbi.2006.07.011

  4. Bargmann BO, Laxalt AM, Riet BT, Testerink C, Merquiol E, Mosblech A, Leon-Reyes A, Pieterse CM, Haring MA, Heilmann I, Bartels D, Munnik T (2009a) Reassessing the role of phospholipase D in the Arabidopsis wounding response. Plant Cell Environ 32:743–757. doi:10.1111/j.1365-3040.2009.01962.x

  5. Bargmann BO, Laxalt AM, Riet BT, van Schooten B, Merquiol E, Testerink C, Haring MA, Bartels D, Munnik T (2009b) Multiple PLDs required for high salinity and water deficit tolerance in plants. Plant Cell Physiol 50:78–89. doi:10.1093/pcp/pcn173

  6. Ben Rejeb K, Abdelly C, Savouré A (2014) How reactive oxygen species and proline face stress together. Plant Physiol Biochem 80:278–284. doi:10.1016/j.plaphy.2014.04.007

  7. Ben Rejeb K, Lefebvre-De Vos D, Le Disquet I, Leprince A-S, Bordenave M, Maldiney R, Jdey A, Abdelly C, Savouré A (2015) Hydrogen peroxide produced by NADPH oxidases increases proline accumulation during salt or mannitol stress in Arabidopsis thaliana. New Phytol 208:1138–1148. doi:10.1111/nph.13550

  8. Beyer WF, Fridovich I (1987) Assaying for superoxide dismutase activity: some large consequences of minor changes in conditions. Anal Biochem 161:559–566. doi:10.1016/0003-2697(87)90489-1

  9. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

  10. Chen MX, Lung SC, Du ZY, Chye ML (2013) Engineering plants to tolerate abiotic stresses. Biocatal Agric Biotechnol 3:81–87. doi:10.1016/j.bcab.2013.09.010

  11. Crowe ML, Serizet C, Thareau V, Aubourg S, Rouzé P, Hilson P, Beynon J, Weisbeek P, van Hummelen P, Reymond P, Paz-Ares J, Nietfeld W, Trick M (2003) CATMA: a complete Arabidopsis GST database. Nucleic Acids Res 31:156–158

  12. Deal SB (1954) Flame photometric determination of sodium and potassium. Anal Chem 26:598–599. doi:10.1021/ac60087a063

  13. Deinlein U, Stephan AB, Horie T, Luo W, Xu G, Schroeder JI (2014) Plant salt-tolerance mechanisms. Trends Plant Sci 19:371–379. doi:10.1016/j.tplants.2014.02.001

  14. Ellouzi H, Ben Hamed K, Cela J, Munné-Bosch S, Abdelly C (2011) Early effects of salt stress on the physiological and oxidative status of Cakile maritima (halophyte) and Arabidopsis thaliana (glycophyte). Physiol Plant 142:128–143. doi:10.1111/j.1399-3054.2011.01450.x

  15. Ellouzi H, Ben Hamed K, Asensi-Fabado MA, Müller M, Abdelly C, Munné-Bosch S (2013) Drought and cadmium may be as effective as salinity in conferring subsequent salt stress tolerance in Cakile maritima. Planta 237:1311–1323. doi:10.1007/s00425-013-1847-7

  16. Galvan-Ampudia CS, Julkowska MM, Darwish E, Gandullo J, Korver RA, Brunoud G, Haring MA, Munnik T, Vernoux T, Testerink C (2013) Halotropism is a response of plant roots to avoid a saline environment. Curr Biol 23:2044–2050. doi:10.1016/j.cub.2013.08.042

  17. Gardiner JC, Harper JD, Weerakoon ND, Collings DA, Ritchie S, Gilroy S, Cyr RJ, Marc J (2001) A 90-kD phospholipase D from tobacco binds to microtubules and the plasma membrane. Plant Cell 13:2143–2158

  18. Ghars MA, Parre E, Debez A, Bordenave M, Richard L, Leport L, Bouchereau A, Savoure A, Abdelly C (2008) Comparative salt tolerance analysis between Arabidopsis thaliana and Thellungiella halophila, with special emphasis on K+/Na+ selectivity and proline accumulation. J Plant Physiol 165:588–599. doi:10.1016/j.jplph.2007.05.014

  19. Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem 48:909–930. doi:10.1016/j.plaphy.2010.08.016

  20. Hewitt EJ (1966) The composition of the nutrient solution. Sand and water culture methods used in the study of plant nutrition. Commonwealth Agricultural Bureaux, Farnham Royal, pp 187–246

  21. Hong Y, Pan X, Welti R, Wang X (2008) Phospholipase Dalpha3 is involved in the hyperosmotic response in Arabidopsis. Plant Cell 20:803–816. doi:10.1105/tpc.107.056390

  22. Hong Y, Zhang W, Wang X (2010) Phospholipase D and phosphatidic acid signalling in plant response to drought and salinity. Plant Cell Environ 33:627–635

  23. Hong J-P, Takeshi Y, Kondou Y, Schachtman DP, Matsui M, Shin R (2013) Identification and characterization of transcription factors regulating Arabidopsis HAK5. Plant Cell Physiol 54:1478–1490. doi:10.1093/pcp/pct094

  24. Hou Q, Ufer G, Bartels D (2016) Lipid signalling in plant responses to abiotic stress. Plant Cell Environ 39:1029–1048. doi:10.1111/pce.12666

  25. Hunt R (1990) Basic Growth analysis: plant growth analysis for beginners. Unwin Hyman, London

  26. Julkowska MM, Testerink C (2015) Tuning plant signaling and growth to survive salt. Trends Plant Sci 20:586–594. doi:10.1016/j.tplants.2015.06.008

  27. Li G, Xue H-W (2007) Arabidopsis PLDzeta2 regulates vesicle trafficking and is required for auxin response. Plant Cell 19:281–295. doi:10.1105/tpc.106.041426

  28. Li M, Qin C, Welti R, Wang X (2006) Double knockouts of phospholipases Dzeta1 and Dzeta2 in Arabidopsis affect root elongation during phosphate-limited growth but do not affect root hair patterning. Plant Physiol 140:761–770. doi:10.1104/pp.105.070995

  29. Li M, Hong Y, Wang X (2009) Phospholipase D- and phosphatidic acid-mediated signaling in plants. Biochim Biophys Acta Mol Cell Biol Lipids 1791:927–935. doi:10.1016/j.bbalip.2009.02.017

  30. Lurin C, Andrés C, Aubourg S, Bellaoui M, Bitton F, Bruyère C, Caboche M, Debast C, Gualberto J, Hoffmann B, Lecharny A, Ret ML, Martin-Magniette M-L, Mireau H et al (2004) Genome-wide analysis of Arabidopsis pentatricopeptide repeat proteins reveals their essential role in organelle biogenesis. Plant Cell 16:2089–2103. doi:10.1105/tpc.104.022236

  31. Magome H, Yamaguchi S, Hanada A, Kamiya Y, Oda K (2004) dwarf and delayed-flowering 1, a novel Arabidopsis mutant deficient in gibberellin biosynthesis because of overexpression of a putative AP2 transcription factor. Plant J 37:720–729

  32. Magome H, Yamaguchi S, Hanada A, Kamiya Y, Oda K (2008) The DDF1 transcriptional activator upregulates expression of a gibberellin-deactivating gene, GA2ox7, under high-salinity stress in Arabidopsis. Plant J 56:613–626. doi:10.1111/j.1365-313X.2008.03627.x

  33. McLoughlin F, Testerink C (2013) Phosphatidic acid, a versatile water-stress signal in roots. Front Plant Sci 4:525. doi:10.3389/fpls.2013.00525

  34. Meijer HJ, Munnik T (2003) Phospholipid-based signaling in plants. Annu Rev Plant Biol 54:265–306

  35. Miller G, Suzuki N, Ciftci-Yilmaz S, Mittler R (2010) Reactive oxygen species homeostasis and signalling during drought and salinity stresses. Plant Cell Environ 33:453–467. doi:10.1111/j.1365-3040.2009.02041.x

  36. Mittler R (2016) ROS are good. Trends Plant Sci 22:11–19. doi:10.1016/j.tplants.2016.08.002

  37. Mollinedo F (2012) Lipid raft involvement in yeast cell growth and death. Front Oncol 2:140

  38. Munns R, Tester M (2008) Mechanisms of salinity tolerance. Annu Rev Plant Biol 59:651–681. doi:10.1146/Annurev.Arplant.59.032607.092911

  39. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue culture. Physiol Plant 15:473–497

  40. Muzi C, Camoni L, Visconti S, Aducci P (2016) Cold stress affects H+-ATPase and phospholipase D activity in Arabidopsis. Plant Physiol Biochem 108:328–336

  41. Ohashi Y, Oka A, Rodrigues-Pousada R, Possenti M, Ruberti I, Morelli G, Aoyama T (2003) Modulation of phospholipid signaling by GLABRA2 in root-hair pattern formation. Science 300:1427–1430

  42. Parre E, Ghars MA, Leprince AS, Thiery L, Lefebvre D, Bordenave M, Richard L, Mazars C, Abdelly C, Savouré A (2007) Calcium signaling via phospholipase C is essential for proline accumulation upon ionic but not nonionic hyperosmotic stresses in Arabidopsis. Plant Physiol 144:503–512. doi:10.1104/pp.106.095281

  43. Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29:e45. doi:10.1093/nar/29.9.e45

  44. Planchais S, Cabassa C, Toka I, Justin A-M, Renou J-P, Savouré A, Carol P (2014) BASIC AMINO ACID CARRIER 2 gene expression modulates arginine and urea content and stress recovery in Arabidopsis leaves. Front Plant Sci 5:330. doi:10.3389/fpls.2014.00330

  45. Qin C, Wang X (2002) The Arabidopsis phospholipase D family. Characterization of a calcium-independent and phosphatidylcholine-selective PLDzeta1 with distinct regulatory domains. Plant Physiol 128:1057–1068

  46. Rangani J, Parida AK, Panda A, Kumari A (2016) Coordinated changes in antioxidative enzymes protect the photosynthetic machinery from salinity induced oxidative damage and confer salt tolerance in an extreme halophyte Salvadora persica L. Front Plant Sci 7:50. doi:10.3389/fpls.2016.00050

  47. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. Cold Spring Harbor Laboratory Press, New York

  48. Singh A, Pandey A, Baranwal V, Kapoor S, Pandey GK (2012) Comprehensive expression analysis of rice phospholipase D gene family during abiotic stress and development. Plant Signal Behav 7:847–855

  49. Slama I, Abdelly C, Bouchereau A, Flowers T, Savouré A (2015) Diversity, distribution and roles of osmoprotective compounds accumulated in halophytes under abiotic stress. Ann Bot 115:433–447. doi:10.1093/aob/mcu239

  50. Thiery L, Leprince AS, Lefebvre D, Ghars MA, Debarbieux E, Savouré A (2004) Phospholipase D is a negative regulator of proline biosynthesis in Arabidopsis thaliana. J Biol Chem 279:14812–14818

  51. Vergnolle C, Vaultier M-N, Taconnat L, Renou J-P, Kader J-C, Zachowski A, Ruelland E (2005) The cold-induced early activation of phospholipase C and D pathways determines the response of two distinct clusters of genes in Arabidopsis cell suspensions. Plant Physiol 139:1217–1233. doi:10.1104/pp.105.068171

  52. Wang X (2005) Regulatory functions of phospholipase D and phosphatidic acid in plant growth, development, and stress responses. Plant Physiol 139:566–573

  53. Yu L, Nie J, Cao C, Jin Y, Yan M, Wang F, Liu J, Xiao Y, Liang Y, Zhang W (2010) Phosphatidic acid mediates salt stress response by regulation of MPK6 in Arabidopsis thaliana. New Phytol 188:762–773

  54. Zhang Y, Zhu H, Zhang Q, Li M, Yan M, Wang R, Wang L, Welti R, Zhang W, Wang X (2009) Phospholipase Dα1 and phosphatidic acid regulate NADPH oxidase activity and production of reactive oxygen species in ABA-mediated stomatal closure in Arabidopsis. Plant Cell 21:2357–2377. doi:10.1105/tpc.108.062992

  55. Zhu JK (2000) Genetic analysis of plant salt tolerance using Arabidopsis. Plant Physiol 124:941–948

  56. Zhu JK (2002) Salt and drought stress signal transduction in plants. Annu Rev Plant Biol 53:247–273

Download references

Acknowledgements

This work was supported by the UPMC (France) and the CBBC (Tunisia) and also supported by the Tunisian-French UTIQUE Network (No. 13G0929). We thank Frederique Bitton and Jean-Pierre Renou from URG-INRA, Evry (France) for help producing the microarray data. Bualuang Faiyue thanks the Junior Research Fellowship Program supported by the French Embassy in Thailand.

Author information

Correspondence to Arnould Savouré.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (PDF 208 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Ben Othman, A., Ellouzi, H., Planchais, S. et al. Phospholipases Dζ1 and Dζ2 have distinct roles in growth and antioxidant systems in Arabidopsis thaliana responding to salt stress. Planta 246, 721–735 (2017). https://doi.org/10.1007/s00425-017-2728-2

Download citation

Keywords

  • Ion relations
  • Phospholipase D
  • PLDζ
  • Reactive oxygen species
  • Salt stress
  • Transcriptome