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Fusaric acid induction of programmed cell death modulated through nitric oxide signalling in tobacco suspension cells

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Abstract

Fusaric acid (FA) is a nonhost-selective toxin mainly produced by Fusarium oxysporum, the causal agent of plant wilt diseases. We demonstrate that FA can induce programmed cell death (PCD) in tobacco suspension cells and the FA-induced PCD is modulated by nitric oxide (NO) signalling. Cells undergoing cell death induced by FA treatment exhibited typical characteristics of PCD including cytoplasmic shrinkage, chromatin condensation, DNA fragmentation, membrane plasmolysis, and formation of small cytoplasmic vacuoles. In addition, caspase-3-like activity was activated upon the FA treatment. The process of FA-induced PCD was accompanied by a rapid accumulation of NO in a FA dose-dependent manner. Pre-treatment of cells with NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO) or NO synthase inhibitor N G-monomethyl-arginine monoacetate (L-NMMA) significantly reduced the rate of FA-induced cell death. Furthermore, the caspase-3-like activity and the expression of PAL and Hsr203J genes were alleviated by application of cPTIO or L-NMMA to FA-treated tobacco cells. This indicates that NO is an important factor involved in the FA-induced PCD. Our results also show that pre-treatment of tobacco cells with a caspase-3-specific inhibitor, Ac-DEVD-CHO, can reduce the rate of FA-induced cell death. These results demonstrate that the FA-induced cell death is a PCD and is modulated by NO signalling through caspase-3-like activation.

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Abbreviations

Ac-DEVD-CHO:

Acetyl-Asp-Glu-Val-Asp-aldehyde

cPTIO:

2-(4-Carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide

DAF-2:

4,5-Diaminofluorescein

DAF-FM DA:

4-Amino-5-methylamino-2′,7′-difluorescein diacetate

DAPI:

4′,6-Diamidino-2-phenylindole

FA:

Fusaric acid

L-NMMA:

N G-monomethyl-arginine monoacetate

NO:

Nitric oxide

PBS:

Phosphate buffered saline

PCD:

Programmed cell death

RT-PCR:

Reverse transcription-polymerase chain reaction

TUNEL:

Terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling

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Acknowledgments

This work was supported by the National Natural Science Foundation (31171806, 30871620), Shandong Provincial Natural Science Foundation (ZR2012CM032), and Anhui Province Tobacco Company Project (20100551002, 20100551005) in China.

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Authors and Affiliations

  1. Department of Plant Pathology, Shandong Agricultural University, Taian, 271018, China

    Jiao Jiao, Xiaoping Zhu & Yuancun Liang

  2. Tobacco Research Institute, Anhui Academy of Agricultural Sciences, Hefei, 230031, China

    Benguo Zhou & Zhengliang Gao

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  1. Jiao Jiao
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  2. Benguo Zhou
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  3. Xiaoping Zhu
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  4. Zhengliang Gao
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  5. Yuancun Liang
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Correspondence to Yuancun Liang.

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J. Jiao and B. Zhou contributed equally to this work.

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Jiao, J., Zhou, B., Zhu, X. et al. Fusaric acid induction of programmed cell death modulated through nitric oxide signalling in tobacco suspension cells. Planta 238, 727–737 (2013). https://doi.org/10.1007/s00425-013-1928-7

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