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Protoplasma

, Volume 250, Issue 1, pp 273–284 | Cite as

Ethylene signaling in salt stress- and salicylic acid-induced programmed cell death in tomato suspension cells

  • Péter Poór
  • Judit Kovács
  • Dóra Szopkó
  • Irma TariEmail author
Original Article

Abstract

Salt stress- and salicylic acid (SA)-induced cell death can be activated by various signaling pathways including ethylene (ET) signaling in intact tomato plants. In tomato suspension cultures, a treatment with 250 mM NaCl increased the production of reactive oxygen species (ROS), nitric oxide (NO), and ET. The 10−3 M SA-induced cell death was also accompanied by ROS and NO production, but ET emanation, the most characteristic difference between the two cell death programs, did not change. ET synthesis was enhanced by addition of ET precursor 1-aminocyclopropane-1-carboxylic acid, which, after 2 h, increased the ROS production in the case of both stressors and accelerated cell death under salt stress. However, it did not change the viability and NO levels in SA-treated samples. The effect of ET induced by salt stress could be blocked with silver thiosulfate (STS), an inhibitor of ET action. STS reduced the death of cells which is in accordance with the decrease in ROS production of cells exposed to high salinity. Unexpectedly, application of STS together with SA resulted in increasing ROS and reduced NO accumulation which led to a faster cell death. NaCl- and SA-induced cell death was blocked by Ca2+ chelator EGTA and calmodulin inhibitor W-7, or with the inhibitors of ROS. The inhibitor of MAPKs, PD98059, and the cysteine protease inhibitor E-64 reduced cell death in both cases. These results show that NaCl induces cell death mainly by ET-induced ROS production, but ROS generated by SA was not controlled by ET in tomato cell suspension.

Keywords

Ethylene Programmed cell death Reactive oxygen species Salicylic acid Salt stress Tomato suspension culture 

Abbreviations

ACC

1-Aminocyclopropane-1-carboxylic acid

AVG

Aminoethoxyvinyl glycine

CAT

Catalase

cPTIO

Carboxyphenyl-tetramethylimidazoline-oxide

DAF-2 DA

4,5 Diaminofluorescein-diacetate

DPI

Diphenyleneiodonium chloride

E-64

N-(trans-epoxysuccinyl)-l-leucine 4-guanidinobutylamide

EGTA

Ethylene glycol tetraacetic acid

EL

Relative electrolyte leakage

ET

Ethylene

FDA

Fluorescein diacetate

H2DCFDA

2′,7′-Dichlorofluorescein diacetate

HR

Hypersensitive response

KOR

Depolarization-active outward-rectifying K+ channels

MAPK

Mitogen-activated protein kinase

NO

Nitric oxide

Nr

Never ripe tomato (ethylene receptor) mutant

NSCC

Non-selective cation channels

PBS

Phosphate-buffered saline

PCD

Programmed cell death

PD98059

Amino-methoxyphenyl-benzopyran

PM

Plasma membrane

ROS

Reactive oxygen species

SA

Salicylic acid

SNP

Sodium nitroprusside

SOD

Superoxide dismutase

STS

Silver thiosulphate

TUNEL

Terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling

W-7

N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide hydrochloride

Notes

Acknowledgments

We thank Kispálné Szabó Ibolya for her excellent technical assistance. This work was supported by grants from the Hungarian National Scientific Research Foundation (OTKA K76854 and OTKA K 101243).

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. Affenzeller MJ, Dareshouri A, Andosch A, Lutz C, Lutz-Meindl U (2009) Salt stress induced cell death in the unicellular green alga Micrasterias denticulata. J Exp Bot 60:939–954PubMedCrossRefGoogle Scholar
  2. Ahlfors R, Brosché M, Kangasjärvi J (2009) Ozone and nitric oxide interaction in Arabidopsis thaliana. Plant Signal Behav 4:878–879PubMedCrossRefGoogle Scholar
  3. Alvarez ME (2000) Salicylic acid in the machinery of hypersensitive cell death and disease resistance. Plant Mol Biol 44:429–442PubMedCrossRefGoogle Scholar
  4. Arfan M, Athar HR, Ashraf M (2007) Does exogenous application of salicylic acid through the rooting medium modulate growth and photosynthetic capacity in two differently adapted spring wheat cultivars under salt stress? J Plant Physiol 164:685–694PubMedCrossRefGoogle Scholar
  5. Bastianelli F, Costa A, Vescovi M, D’Apuzzo E, Zottini M, Chiurazzi M, Schiavo FL (2010) Salicylic acid differentially affects suspension cell cultures of Lotus japonicus and one of its non-symbiotic mutants. Plant Mol Biol 72:469–483PubMedCrossRefGoogle Scholar
  6. Bi YH, Chen WL, Zhang WN, Zhou Q, Yun LJ, Xing D (2009) Production of reactive oxygen species, impairment of photosynthetic function and dynamic changes in mitochondria are early events in cadmium-induced cell death in Arabidopsis thaliana. Biol Cell 101:629–643PubMedCrossRefGoogle Scholar
  7. Bleecker AB, Kende H (2000) Ethylene: a gaseous signal molecule in plants. Annu Rev Cell Dev Biol 16:1–18PubMedCrossRefGoogle Scholar
  8. Byczkowska A, Kunikowska A, Kaźmierczak A (2012) Determination of ACC-induced cell-programmed death in roots of Vicia faba ssp. minor seedlings by acridine orange and ethidium bromide staining. Protoplasma. doi: 10.1007/s00709-0120383-9
  9. Clarke A, Desikan R, Hurst RD, Hancock JT, Neil SJ (2000) NO way back: nitric oxide and programmed cell death in Arabidopsis thaliana suspension cultures. Plant J 24:667–677PubMedCrossRefGoogle Scholar
  10. Dangl JL, Dietrich RA, Thomas H (2000) Senescence and programmed cell death. Biochem Mol Biol Plants 2000:1044–1100Google Scholar
  11. Danon A, Delorme V, Mailhac N, Gallois P (2000) Plant programmed cell death: a common way to die. Plant Physiol Biochem 38:647–655CrossRefGoogle Scholar
  12. De Jong A, Hoeberichts FA, Yakimova ET, Maximova E, Woltering EJ (2000) Chemical-induced apoptotic cell death in tomato cells: involvement of caspase-like proteases. Planta 211:656–662PubMedCrossRefGoogle Scholar
  13. De Jong A, Yakimova ET, Kapchina VM, Woltering EJ (2002) A critical role for ethylene in hydrogen peroxide release during programmed cell death in tomato suspension cells. Planta 21:537–545CrossRefGoogle Scholar
  14. del Pozo O, Pedley KF, Martin GB (2004) MAPKKKa is a positive regulator of cell death associated with both plant immunity and disease. EMBO J 23:3072–3082PubMedCrossRefGoogle Scholar
  15. Delledonne M, Xia Y, Dixon RA, Lamb C (1998) Nitric oxide functions as a signal in plant disease resistance. Nature 394:585–588PubMedCrossRefGoogle Scholar
  16. Duan Y, Zhang W, Li B, Wang Y, Li K, Sodmergen HC, Zhang Y, Li X (2010) An endoplasmic reticulum response pathway mediates programmed cell death of root tip induced by water stress in Arabidopsis. New Phytol 186:681–695PubMedCrossRefGoogle Scholar
  17. Ederli L, Morettini R, Borgogni A, Wasternack C, Miersch O, Reale L, Ferranti F, Tosti N, Pasqualini S (2006) Interaction between nitric oxide and ethylene in the induction of alternative oxidase in ozone-treated tobacco plants. Plant Physiol 142:595–608PubMedCrossRefGoogle Scholar
  18. Gamborg O, Miller R, Ojima K (1968) Nutrient requirement suspensions cultures of soybean root cells. Exp Cell Res 50:151–158PubMedCrossRefGoogle Scholar
  19. García-Heredia JM, Hervás M, De la Rosa MA, Navarro JA (2008) Acetylsalicylic acid induces programmed cell death in Arabidopsis cell cultures. Planta 228:89–97PubMedCrossRefGoogle Scholar
  20. Gechev TS, Hille J (2005) Hydrogen peroxide as a signal controlling plant programmed cell death. J Cell Biol 168:17–20PubMedCrossRefGoogle Scholar
  21. Gémes K (2011) Improving salt stress acclimation of tomato by salicylic acid: the role of reactive oxygen species and nitric oxide. PhD Thesis, (in Hungarian) pp. 63–66Google Scholar
  22. Gémes K, Poór P, Horváth E, Zs K, Szopkó D, Szepesi Á, Tari I (2011) Cross-talk between salicylic acid and NaCl-generated reactive oxygen species and nitric oxide in tomato during acclimation to high salinity. Physiol Plant 142:179–192PubMedCrossRefGoogle Scholar
  23. Gunawardena A, Pearce DM, Jackson MB, Hawes CR, Evans DE (2001) Characterisation of programmed cell death during aerenchyma formation induced by ethylene or hypoxia in roots of maize (Zea mays L.). Planta 212:205–214PubMedCrossRefGoogle Scholar
  24. Hoeberichts FA, ten Have A, Woltering EJ (2003) A tomato metacaspase gene is upregulated during programmed cell death in Botrytis cinerea-infected leaves. Planta 217:517–522PubMedCrossRefGoogle Scholar
  25. Huh GH, Damaz B, Matsumoto TK, Reddy MP, Rus AM, Ibeas JI (2002) Salt causes ion disequilibrium-induced programmed cell death in yeast and plants. Plant J 29:649–659PubMedCrossRefGoogle Scholar
  26. Jones ML, Chaffin GS, Eason JR, Clark DG (2005) Ethylene-sensitivity regulates proteolytic activity and cysteine protease gene expression in petunia corollas. J Exp Bot 56:2733–2744PubMedCrossRefGoogle Scholar
  27. Joseph B, Jini D (2010) Salinity induced programmed cell death in plants: challenges and opportunities for salt-tolerant plants. J Plant Sci 5:376–390CrossRefGoogle Scholar
  28. Kubis SE, Castilho AMMF, Vershinin AV, Heslop-Harrison JS (2003) Retroelements, transposons and methylation status in the genome of oil palm (Elaeis guineensis) and the relationship to somaclonal variation. Plant Mol Biol 52:69–79PubMedCrossRefGoogle Scholar
  29. Kumar V, Parvatam G, Ravishankar GA (2009) AgNO3—a potential regulator of ethylene activity and plant growth modulator. Electric J Biotechnol 12:1–15Google Scholar
  30. Lam E, Kato N, Lawton M (2001) Programmed cell death, mitochondria and plant hypersensitive response. Nature 411:848–853PubMedCrossRefGoogle Scholar
  31. Leslie CA, Romani RJ (1986) Salicylic acid: a new inhibitor of ethylene biosynthesis. Plant Cell Rep 5:144–146CrossRefGoogle Scholar
  32. Love AJ, Milner JJ, Sadanandom A (2008) Timing is everything: regulatory overlap in plant cell death. Trends Plant Sci 13:589–595PubMedCrossRefGoogle Scholar
  33. Mittler R, Vanderauwera S, Gollery M, Breusegem F (2004) Reactive oxygen gene network of plants. Trends Plant Sci 9:490–498PubMedCrossRefGoogle Scholar
  34. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  35. Overmyer K, Brosché M, Kangasjärvi J (2003) Reactive oxygen species and hormonal control of cell death. Trends Plant Sci 8:335–342PubMedCrossRefGoogle Scholar
  36. Poór P, Tari I (2011) Ethylene-regulated reactive oxygen species and nitric oxide under salt stress in tomato cell suspension culture. Acta Biol Szeged 55:143–146Google Scholar
  37. Poór P, Gémes K, Horváth F, Szepesi Á, Simon ML, Tari I (2011a) Salicylic acid treatment via the rooting medium interferes with the stomatal response, CO2 fixation rate and carbohydrate metabolism in tomato and decreases the harmful effects of subsequent salt stress. Plant Biol 13:105–114PubMedCrossRefGoogle Scholar
  38. Poór P, Szopkó D, Tari I (2011b) Ionic homeostasis disturbance is involved in tomato cell death induced by NaCl and salicylic acid. In Vitro Cell Dev Biol Plant. doi: 10.1007/s11627-011-9419-7, #IVPL-D-10-00389R2
  39. Quirino BF, Noh YS, Himelblau E, Amasino RM (2000) Molecular aspects of leaf senescence. Trends Plant Sci 5:278–282PubMedCrossRefGoogle Scholar
  40. Rao MV, Lee H, Davis KR (2002) Ozone-induced ethylene production is dependent on salicylic acid, and both salicylic acid and ethylene act in concert to regulate ozone-induced cell death. Plant J 32:447–456PubMedCrossRefGoogle Scholar
  41. Rogers HJ (2005) Cell death and organ development in plants. Curr Top Dev Biol 71:225–261PubMedCrossRefGoogle Scholar
  42. Shabala S (2009) Salinity and programmed cell death: unravelling mechanisms for ion specific signalling. J Exp Bot 60:709–712PubMedCrossRefGoogle Scholar
  43. Shi Q, Bao Z, Zhu Z, Ying Q, Qian Q (2006) Effects of different treatments of salicylic acid on heat tolerance, chlorophyll fluorescence, and antioxidant enzyme activity in seedlings of Cucumis sativa L. Plant Growth Regul 48:127–135CrossRefGoogle Scholar
  44. Siddiqui MH, Al-Whaibi MH, Basalah MO (2011) Role of nitric oxide in tolerance of plants to abiotic stress. Protoplasma 248:447–455PubMedCrossRefGoogle Scholar
  45. Sun J, Li L, Liu M, Wang M, Ding M, Deng S, Lu C, Zhou X, Shen X, Zheng X, Chen S (2010) Hydrogen peroxide and nitric oxide mediate K+/Na+ homeostasis and antioxidant defense in NaCl-stressed callus cells of two contrasting poplars. Plant Cell Tissue Organ Cult 103:205–215CrossRefGoogle Scholar
  46. Szepesi Á, Csiszár J, Gémes K, Horváth E, Horváth F, Simon ML, Tari I (2009) Salicylic acid improves acclimation to salt stress by stimulating abscisic aldehyde oxidase activity and abscisic acid accumulation, and increases Na+ content in leaves without toxicity symptoms in Solanum lycopersicum L. J Plant Physiol 166:914–925PubMedCrossRefGoogle Scholar
  47. Tari I, Csiszár J, Gémes K, Szepesi Á (2006) Modulation of Cu2+ accumulation by (aminoethoxyvinyl)glycine and methylglyoxal bis(guanylhydrazone), the inhibitors of stress ethylene and polyamine synthesis in wheat genotypes. Cereal Res Commun 34:989–996CrossRefGoogle Scholar
  48. Tari I, Poór P, Gémes K (2011) Sublethal concentrations of salicylic acid decrease the formation of reactive oxygen species but maintain an increased nitric oxide production in the root apex of the ethylene-insensitive Never ripe tomato mutants. Plant Signal Behav 6:1263–1266PubMedCrossRefGoogle Scholar
  49. Trobacher CP, Senatore A, Greenwood JS (2006) Masterminds or minions? Cysteine proteinases in plant programmed cell death. Can J Bot-Rev 84:651–667CrossRefGoogle Scholar
  50. van Doorn (2011) Classes of programmed cell death in plants, compared to those in animals. J Exp Bot 62:4749–4761PubMedCrossRefGoogle Scholar
  51. Vartapetian AB, Tuzhikov AL, Chichkova NV, Taliansky M, Wolpert TJ (2011) A plant alternative to animal caspases: subtilisin-like proteases. Cell Death Diff 18:1289–1297CrossRefGoogle Scholar
  52. Wang H, Liang X, Wan Q, Wang X, Bi Y (2009) Ethylene and nitric oxide are involved in maintain ion homeostasis in Arabidopsis callus under salt stress. Planta 230:293–307PubMedCrossRefGoogle Scholar
  53. Wang H, Liang X, Huang J, Zhang D, Lu H, Liu Z, Bi Y (2010a) Involvement of ethylene and hydrogen peroxide in induction of alternative respiratory pathway in salt-treated Arabidopsis calluses. Plant Cell Physiol 51:1754–1765PubMedCrossRefGoogle Scholar
  54. Wang J, Li X, Liu Y, Zhao X (2010b) Salt stress induces programmed cell death in Thellungiella halophila suspension-cultured cells. J Plant Physiol 167:1145–1151PubMedCrossRefGoogle Scholar
  55. Wi SJ, Jang SJ, Park KY (2010) Inhibition of biphasic ethylene production enhances tolerance to abiotic stress by reducing the accumulation of reactive oxygen species in Nicotiana tabacum. Mol Cells 30:37–49PubMedCrossRefGoogle Scholar
  56. Woltering EJ (2004) Death proteases come alive. Trends Plant Sci 9:469–472PubMedCrossRefGoogle Scholar
  57. Yakimova ET, Kapchina-Toteva VM, Laarhoven LJ, Harren FM, Woltering EJ (2006) Involvement of ethylene and lipid signalling in cadmium-induced programmed cell death in tomato suspension cells. Plant Physiol Biochem 44:581–589PubMedCrossRefGoogle Scholar
  58. Yakimova ET, Woltering EJ, Kapchina-Toteva VM, Harren FJM, Cristescu SM (2008) Cadmium toxicity in cultured tomato cells-role of ethylene, proteases and oxidative stress in cell death signaling. Cell Biol Int 32:1521–1529CrossRefGoogle Scholar
  59. Zhang S, Klessing DF (2001) MAPK cascades in plant defense signaling. Trends Plant Sci 6:520–527PubMedCrossRefGoogle Scholar
  60. Zottini M, Costa A, Michele RD, Ruzzene M, Carimi F, Schiavo FL (2007) Salicylic acid activates nitric oxide synthesis in Arabidopsis. J Exp Bot 58:1397–1405PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Péter Poór
    • 1
  • Judit Kovács
    • 1
  • Dóra Szopkó
    • 1
  • Irma Tari
    • 1
    Email author
  1. 1.Department of Plant BiologyUniversity of SzegedSzegedHungary

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