Plant Growth Regulation

, Volume 63, Issue 3, pp 207–216 | Cite as

MEK/ERK inhibitor U0126 enhanced salt stress-induced programmed cell death in Thellungiella halophila suspension-cultured cells

  • Jin Wang
  • Xinrong Li
  • Yubing Liu
  • Xin Zhao
  • Cuiyun Chen
  • Fang Tian
Original Research

Abstract

Programmed cell death (PCD) is an active cellular suicide that occurs both in animals and plants throughout development and in response to abiotic or biotic stress. In contrast to plant hypersensitive response-like cell death, little is known about the molecular machinery that regulates the halophyte plant PCD under high salinity stress. Since mitogen-activated protein kinases (MAPKs) are involved in plant response/tolerance to salt stress, and plant MAPK genes belong to the extracellular signal-regulated kinase (ERK) subfamily, we have investigated the role of ERK-like enzymes in high salinity stress-induced cell death in Thellungiella halophila. The data showed that ERK-like enzymes were early (10 min) and transiently activated under 300 mM NaCl stress. Pretreatment with 10 μM U0126, a special MEK/ERK inhibitor, resulted in a small but statistically significant increase of the percentage of terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL)-positive nuclei in contrast to salt alone. The effects of U0126 on H2O2 production and cytochrome c (cyt c) release were also investigated. We found that the pretreatment with U0126 accelerated H2O2 production as well as cyt c release, and eventually enhanced cell death. The results suggest that ERK-like enzymes in Thellungiella halophila may act as a positive regulator of salt tolerance, as illustrated by pretreatment with U0126 which enhanced cell death under high salinity stress.

Keywords

Apoptosis Cytochrome c Hydrogen peroxide Mitogen-activated protein kinase Salt stress 

Notes

Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China (nos. 40825001 and 30870425). The authors are grateful to Rui Li from the School of Basic Medical Sciences of Lanzhou University for his help with the TUNEL assay.

References

  1. Ahlfors R, Macioszek V, Rudd J, Brosché M, Schlichting R, Scheel D, Kangasjärvi J (2004) Stress hormone-independent activation and nuclear translocation of mitogen-activated protein kinases in Arabidopsis thaliana during ozone exposure. Plant J 40:512–522PubMedCrossRefGoogle Scholar
  2. Amtmann A, Bohnert HJ, Bressan RA (2005) Abiotic stress and plant genome evolution. Search for new models. Plant Physiol 138:127–130PubMedCrossRefGoogle Scholar
  3. Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–399PubMedCrossRefGoogle Scholar
  4. Asai T, Tena G, Plotnikova J, Willmann MR, Chiu WL, Gomez-Gomez L, Boller T, Ausubel FM, Sheen J (2002) MAP kinase signalling cascade in Arabidopsis innate immunity. Nature 415:977–983PubMedCrossRefGoogle Scholar
  5. Ashraf M (1999) Breeding for salinity tolerance in plants. Crit Rev Plant Sci 13:17–42Google Scholar
  6. Balk J, Leaver CJ, McCabe PF (1999) Translocation of cytochrome c from the mitochondria to the cytosol occurs during heat-induced programmed cell death in cucumber plants. FEBS Lett 463:151–154PubMedCrossRefGoogle Scholar
  7. 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 72:248–254PubMedCrossRefGoogle Scholar
  8. Bressan RA, Zhang C, Zhang H, Hasegawa PM, Bohnert HJ, Zhu JK (2001) Learning from the Arabidopsis experience: the next gene search paradigm. Plant Physiol 127:1354–1360PubMedCrossRefGoogle Scholar
  9. Casolo V, Petrussa E, Krajnáková J, Macrì F, Vianello A (2005) Involvement of the mitochondrial K(+)ATP channel in H2O2- or NO-induced programmed death of soybean suspension cell cultures. J Exp Bot 56:997–1006PubMedCrossRefGoogle Scholar
  10. Chen PY, Lee KT, Chi WC, Hirt H, Chang CC, Huang HJ (2008) Possible involvement of MAP kinase pathways in acquired metal-tolerance induced by heat in plants. Planta 228:499–509PubMedCrossRefGoogle Scholar
  11. Chinnusamy V, Zhu JK (2003) Plant salt tolerance. In: Hirt H, Shinozaki K (eds) Plant responses to abiotic stress, vol 4. Springer-Verlag, Berlin, Germany, pp 241–270Google Scholar
  12. Chinnusamy V, Schumaker K, Zhu JK (2004) Molecular genetic perspectives on cross-talk and specificity in abiotic stress signalling in plants. J Exp Bot 55:225–236PubMedCrossRefGoogle Scholar
  13. Colcombet J, Hirt H (2008) Arabidopsis MAPKs: a complex signaling network involved in multiple biological processes. Biochem J 413:217–226PubMedCrossRefGoogle Scholar
  14. Cross TG, Scheel-Toellner D, Henriquez NV, Deacon E, Salmon M, Lord JM (2000) Serine/threonine protein kinases and apoptosis. Exp Cell Res 256:34–41PubMedCrossRefGoogle Scholar
  15. de Pinto MC, Francis D, De Gara L (1999) The redox state of the ascorbate-dehydroascorbate pair as a specific sensor of cell division in tobacco BY-2 cells. Protoplasma 209:90–97PubMedCrossRefGoogle Scholar
  16. de Pinto MC, Paradiso A, Leonetti P, De Gara L (2006) Hydrogen peroxide, nitric oxide and cytosolic ascorbate peroxidase at the crossroad between defence and cell death. Plant J 48:784–795PubMedCrossRefGoogle Scholar
  17. Desikan R, Clarke A, Hancock JT, Neill SJ (1999) H2O2 activates a MAP kinase-like enzyme in Arabidopsis thaliana suspension cultures. J Exp Bot 50:1863–1866CrossRefGoogle Scholar
  18. Doyle SM, Diamond M, McCabe PF (2010) Chloroplast and reactive oxygen species involvement in apoptotic-like programmed cell death in Arabidopsis suspension cultures. J Exp Bot 61:473–482PubMedCrossRefGoogle Scholar
  19. Droillard M, Boudsocq M, Barbier-Brygoo H, Laurière C (2002) Different protein kinase families are activated by osmotic stresses in Arabidopsis thaliana cell suspensions: involvement of the MAP kinases AtMPK3 and AtMPK6. FEBS Lett 527:43–50PubMedCrossRefGoogle Scholar
  20. Horbinski C, Chu CT (2005) Kinase signaling cascades in the mitochondrion: a matter of life or death. Free Radic Biol Med 38:2–11PubMedCrossRefGoogle Scholar
  21. Huang Y, Li H, Gupta R, Morris PC, Luan S, Kieber JJ (2000) ATMPK4, an Arabidopsis homolog of mitogen-activated protein kinase, is activated in vitro by AtMEK1 through threonine phosphorylation. Plant Physiol 122:1301–1310PubMedCrossRefGoogle Scholar
  22. Huang L, Li B, Li W, Guo H, Zou F (2009) ATP-sensitive potassium channels control glioma cells proliferation by regulating ERK activity. Carcinogenesis 30:737–744PubMedCrossRefGoogle Scholar
  23. Ichimura K, Mizoguchi T, Irie K, Morris P, Giraudat J, Matsumoto K, Shinozaki K (1998) Isolation of ATMEKK1 (a MAP kinase kinase kinase)-interacting proteins and analysis of a MAP kinase cascade in Arabidopsis. Biochem Biophys Res Comm 253:532–543PubMedCrossRefGoogle Scholar
  24. Ichimura K, Mizoguchi T, Yoshida R, Yuasa T, Shinozaki K (2000) Various abiotic stresses rapidly activate Arabidopsis MAP kinases ATMPK4 and ATMPK6. Plant J 24:655–665PubMedCrossRefGoogle Scholar
  25. Inan G, Zhang Q, Li P, Wang Z, Cao Z, Zhang H, Zhang C, Quist TM, Goodwin SM, Zhu J, Shi H, Damsz B, Charbaji T, Gong Q, Ma S, Fredricksen M, Galbraith DW, Jenks MA, Rhodes D, Hasegawa PM, Bohnert HJ, Joly RJ, Bressan RA, Zhu JK (2004) Salt cress: a halophyte and cryophyte Arabidopsis relative model system and its applicability to molecular genetic analyses of growth and development of extremophiles. Plant Physiol 135:1718–1737PubMedCrossRefGoogle Scholar
  26. Kim M, Ahn JW, Jin UH, Choi D, Paek KH, Pai HS (2003) Activation of the programmed cell death pathway by inhibition of proteasome function in plants. J Biol Chem 278:19406–19415PubMedCrossRefGoogle Scholar
  27. Kovtun Y, Chiu WL, Tena G, Sheen J (2000) Functional analysis of oxidative stress-activated mitogen-activated protein kinase cascade in plants. Proc Natl Acad Sci USA 97:2940–2945PubMedCrossRefGoogle Scholar
  28. Kroj T, Rudd JJ, Nürnberger T, Gäbler Y, Lee J, Scheel D (2003) Mitogen-activated protein kinases play an essential role in oxidative burst-independent expression of pathogenesis-related genes in parsley. J Biol Chem 278:2256–2264PubMedCrossRefGoogle Scholar
  29. Laloi C, Apel K, Danon A (2004) Reactive oxygen signalling: the latest news. Curr Opin Plant Biol 7:323–328PubMedCrossRefGoogle Scholar
  30. Lee J, Klessig DF, Nürnberger T (2001) A harpin binding site in tobacco plasma membranes mediates activation of the pathogenesis-related gene HIN1 independent of extracellular calcium but dependent on mitogen-activated protein kinase activity. Plant Cell 13:1079–1093PubMedCrossRefGoogle Scholar
  31. Levine A, Tenhaken R, Dixon R, Lamb C (1994) H2O2 from the oxidative burst orchestrates the plant hypersensitive disease resistance response. Cell 79:583–593PubMedCrossRefGoogle Scholar
  32. Lew RR, Levina NN, Shabala L, Anderca MI, Shabala SN (2006) Role of a mitogen-activated protein kinase cascade in ion flux-mediated turgor regulation in fungi. Eukaryot Cell 5:480–487PubMedCrossRefGoogle Scholar
  33. Li S, Šamaj J, Franklin-Tong VE (2007) A mitogen-activated protein kinase signals to programmed cell death induced by self-incompatibility in Papaver Pollen. Plant Physiol 145:236–245PubMedCrossRefGoogle Scholar
  34. Lin J, Wang Y, Wang G (2006) Salt stress-induced programmed cell death in tobacco protoplasts is mediated by reactive oxygen species and mitochondrial permeability transition pore status. J Plant Physiol 163:731–739PubMedCrossRefGoogle Scholar
  35. Liu Y, Ren D, Pike S, Pallardy S, Gassmann W, Zhang S (2007) Chloroplast-generated reactive oxygen species are involved in hypersensitive response-like cell death mediated by a mitogen-activated protein kinase cascade. Plant J 51:941–954PubMedCrossRefGoogle Scholar
  36. Maathuis FJM, Amtmann A (1999) K+ nutrition and Na+ toxicity: the basis of cellular K+/Na+ ratios. Ann Bot 84:123–133CrossRefGoogle Scholar
  37. MAPK Group (2002) Mitogen-activated protein kinase cascades in plants: a new nomenclature. Trends Plant Sci 7:301–308CrossRefGoogle Scholar
  38. Mizoguchi T, Irie K, Hirayama T, Hayashida N, Yamaguchi-Shinozaki K, Matsumoto K, Shinozaki K (1996) A gene encoding a mitogen-activated protein kinase kinase kinase is induced simultaneously with genes for a mitogen-activated protein kinase and an S6 ribosomal protein kinase by touch, cold, and water stress in Arabidopsis thaliana. Proc Natl Acad Sci USA 93:765–769PubMedCrossRefGoogle Scholar
  39. Mlejnek P, Procházka S (2002) Activation of caspase-like proteases and induction of apoptosis by isopentenyladenosine in tobacco BY-2 cells. Planta 215:158–166PubMedCrossRefGoogle Scholar
  40. Moon H, Lee B, Choi G, Shin D, Prasad DT, Lee O, Kwak SS, Kim DH, Nam J, Bahk J, Hong JC, Lee SY, Cho MJ, Lim CO, Yun DJ (2003) NDP kinase 2 interacts with two oxidative stress-activated MAPKs to regulate cellular redox state and enhances multiple stress tolerance in transgenic plants. Proc Natl Acad Sci USA 100:358–363PubMedCrossRefGoogle Scholar
  41. Morris PC (2001) MAP kinase signal transduction pathways in plants. New Phytol 151:67–89CrossRefGoogle Scholar
  42. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473–497CrossRefGoogle Scholar
  43. Pedley KF, Martin GB (2005) Role of mitogen-activated protein kinases in plant immunity. Curr Opin Plant Biol 8:541–547PubMedCrossRefGoogle Scholar
  44. Petersen M, Brodersen P, Naested H, Andreasson E, Lindhart U, Johansen B, Nielsen HB, Lacy M, Austin MJ, Parker JE, Sharma SB, Klessig DF, Martienssen R, Mattsson O, Jensen AB, Mundy J (2000) Arabidopsis MAP kinase 4 negatively regulates systemic acquired resistance. Cell 103:1111–1120PubMedCrossRefGoogle Scholar
  45. Pitzschke A, Djamei A, Bitton F, Hirt H (2009a) A major role of the MEKK1–MKK1/2–MPK4 pathway in ROS signalling. Mol Plant 2:120–137PubMedCrossRefGoogle Scholar
  46. Pitzschke A, Schikora A, Hirt H (2009b) MAPK cascade signalling networks in plant defence. Curr Opin Plant Biol 12:421–426PubMedCrossRefGoogle Scholar
  47. Raman M, Cobb MH (2003) MAP kinase modules: many roads home. Curr Biol 13:R886–R888PubMedCrossRefGoogle Scholar
  48. Ren D, Yang KY, Li GJ, Liu Y, Zhang S (2006) Activation of Ntf4, a tobacco mitogen-activated protein kinase, during plant defense response and its involvement in hypersensitive response-like cell death. Plant Physiol 141:1482–1493PubMedCrossRefGoogle Scholar
  49. Rudd JJ, Keon J, Hammond-Kosack KE (2008) The wheat mitogen-activated protein kinases TaMPK3 and TaMPK6 are differentially regulated at multiple levels during compatible disease interactions with Mycosphaerella graminicola. Plant Physiol 147:802–815PubMedCrossRefGoogle Scholar
  50. Schwacke R, Hager A (1992) Fungal elicitors induce a transient release of active oxygen species from cultured spruce cells that are dependent on Ca2+ and protein-kinase activity. Planta 187:136–141CrossRefGoogle Scholar
  51. Shabala S (2003) Regulation of potassium transport in leaves: from molecular to tissue level. Ann Bot 92:627–634PubMedCrossRefGoogle Scholar
  52. Shabala S (2009) Salinity and programmed cell death: unravelling mechanisms for ion specific signaling. J Exp Bot 60:709–712PubMedCrossRefGoogle Scholar
  53. Shabala S, Cuin TA (2008) Potassium transport and plant salt tolerance. Physiol Plant 133:651–669PubMedCrossRefGoogle Scholar
  54. Shabala S, Cuin TA, Prismall L, Nemchinov LG (2007) Expression of animal CED-9 anti-apoptotic gene in tobacco modifies plasma membrane ion fluxes in response to salinity and oxidative stress. Planta 227:189–197PubMedCrossRefGoogle Scholar
  55. Taji T, Seki M, Satou M, Sakurai T, Kobayashi M, Ishiyama K, Narusaka Y, Narusaka M, Zhu JK, Shinozaki K (2004) Comparative genomics in salt tolerance between Arabidopsis and Arabidopsis-related halophyte salt cress using Arabidopsis microarray. Plant Physiol 135:1697–1709PubMedCrossRefGoogle Scholar
  56. Teige M, Scheikl E, Eulgem T, Doczi R, Ichimura K, Shinozaki K, Dangl JL, Hirt H (2004) The MKK2 pathway mediates cold and salt stress signaling in Arabidopsis. Mol Cell 15:141–152PubMedCrossRefGoogle Scholar
  57. Tena G, Asai T, Chiu WL, Sheen J (2001) Plant mitogen-activated protein kinase signaling cascades. Curr Opin Plant Biol 4:392–400PubMedCrossRefGoogle Scholar
  58. Tester M, Davenport R (2003) Na+ tolerance and Na+ transport in higher plants. Ann Bot 91:503–527PubMedCrossRefGoogle Scholar
  59. Vacca RA, de Pinto MC, Valenti D, Passarella S, Marra E, De Gara L (2004) Production of reactive oxygen species, alteration of cytosolic ascorbate peroxidase, and impairment of mitochondrial metabolism are early events in heat shock-induced programmed cell death in tobacco Bright-Yellow 2 cells. Plant Physiol 134:1100–1112PubMedCrossRefGoogle Scholar
  60. Vacca RA, Valenti D, Bobba A, Merafina RS, Passarella S, Marra E (2006) Cytochrome c is released in a reactive oxygen species-dependent manner and is degraded via caspase-like proteases in tobacco Bright-Yellow 2 cells en route to heat shock-induced cell death. Plant Physiol 141:208–219PubMedCrossRefGoogle Scholar
  61. Van Breusegem F, Dat JF (2006) Reactive oxygen species in plant cell death. Plant Physiol 141:384–390PubMedCrossRefGoogle Scholar
  62. Vaux DL, Strasser A (1996) The molecular biology of apoptosis. Proc Natl Acad Sci USA 93:2239–2244PubMedCrossRefGoogle Scholar
  63. Wang J, Li X, Liu Y, Zhao X (2010) Salt stress induces programmed cell death in Thellungiella halophila suspension-cultured cells. J Plant Physiol 167:1145–1151PubMedCrossRefGoogle Scholar
  64. Xie Z, Chen Z (2000) Harpin-induced hypersensitive cell death is associated with altered mitochondrial functions in tobacco cells. Mol Plant Microbe Interact 13:183–190PubMedCrossRefGoogle Scholar
  65. Yang KY, Liu Y, Zhang S (2001) Activation of a mitogen-activated protein kinase pathway is involved in disease resistance in tobacco. Proc Natl Acad Sci USA 98:741–746PubMedCrossRefGoogle Scholar
  66. Zhang S, Klessig DF (2001) MAPK cascades in plant defense signaling. Trends Plant Sci 6:520–527PubMedCrossRefGoogle Scholar
  67. Zhang S, Liu Y, Klessig DF (2000) Multiple levels of tobacco WIPK activation during the induction of cell death by fungal elicitins. Plant J 23:339–347PubMedCrossRefGoogle Scholar
  68. Zhang T, Liu Y, Yang T, Zhang L, Xu S, Xue L, An L (2006) Diverse signals converge at MAPK cascades in plant. Plant Physiol Biochem 44:274–283PubMedCrossRefGoogle Scholar
  69. Zhao X, Kim Y, Park G, Xu JR (2005) A mitogen-activated protein kinase cascade regulating infection-related morphogenesis in Magnaporthe grisea. Plant Cell 17:1317–1329PubMedCrossRefGoogle Scholar
  70. Zhu JK (2000) Genetic analysis of plant salt tolerance using Arabidopsis. Plant Physiol 124:941–948PubMedCrossRefGoogle Scholar
  71. Zhu J, Fu X, Koo YD, Zhu JK, Jenney FE Jr, Adams MWW, Zhu Y, Shi H, Yun DJ, Hasegawa PM, Bressan RA (2007) An enhancer mutant of Arabidopsis salt overly sensitive 3 mediates both ion homeostasis and the oxidative stress response. Mol Cell Biol 27:5214–5224PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Jin Wang
    • 1
  • Xinrong Li
    • 1
  • Yubing Liu
    • 1
  • Xin Zhao
    • 1
  • Cuiyun Chen
    • 1
  • Fang Tian
    • 1
  1. 1.Extreme Stress Resistance and Biotechnology Laboratory, Shapotou Desert Research and Experiment StationCold and Arid Regions Environmental and Engineering Research Institute, Chinese Academy of SciencesLanzhouChina

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