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

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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.

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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

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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).

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Plant Biology, University of Szeged, Középfasor 52, 6701, Szeged, P.O. Box 654, Hungary

    Péter Poór, Judit Kovács, Dóra Szopkó & Irma Tari

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  1. Péter Poór
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  2. Judit Kovács
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  3. Dóra Szopkó
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  4. Irma Tari
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Correspondence to Irma Tari.

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Poór, P., Kovács, J., Szopkó, D. et al. Ethylene signaling in salt stress- and salicylic acid-induced programmed cell death in tomato suspension cells. Protoplasma 250, 273–284 (2013). https://doi.org/10.1007/s00709-012-0408-4

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