Accepted Manuscript Title: Hydrogen peroxide production and mitochondrial dysfunction contribute to the fusaric acid-induced programmed cell death in tobacco cells Author: Jiao Jiao Ling Sun Benguo Zhou Zhengliang Gao Yu Hao Xiaoping Zhu Yuancun Liang PII: DOI: Reference:
S0176-1617(14)00087-X http://dx.doi.org/doi:10.1016/j.jplph.2014.03.015 JPLPH 51923
To appear in: Received date: Revised date: Accepted date:
18-10-2013 8-3-2014 19-3-2014
Please cite this article as: Jiao J, Sun L, Zhou B, Gao Z, Hao Y, Zhu X, Liang Y, Hydrogen peroxide production and mitochondrial dysfunction contribute to the fusaric acid-induced programmed cell death in tobacco cells, Journal of Plant Physiology (2014), http://dx.doi.org/10.1016/j.jplph.2014.03.015 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
JIAO J et al
J Plant Physiol
Hydrogen peroxide production and mitochondrial dysfunction contribute to the fusaric acid-induced programmed cell death in tobacco cells Jiao Jiaoa,1, Ling Suna,1, Benguo Zhoub,1, Zhengliang Gaob, Yu Haoa, Xiaoping Zhua, Yuancun
ip t
5
a
cr
Lianga*
Department of Plant Pathology,
10
b
us
Shandong Agricultural University, Taian 271018, Shandong Province, China Tobacco Research Institute,
an
Anhui Academy of Agricultural Sciences, Hefei 23003, Anhui Province, China
Abbreviations: APX, ascorbate peroxidase; AsA, ascorbic acid; CAT, catalase; CsA, cyclosporine
15
M
A; DHR123, dihydrohodamine123; DPI, diphenyl iodonium; FA, fusaric acid; H2O2, hydrogen peroxide; MDA, malondialdehyde; MPTP, mitochondrial permeability transition pore; NO, nitric
te
death.
d
oxide; PBS, phosphate buffered saline; ROS, reactive oxygen species; PCD, programmed cell
20
Ac ce p
* Corresponding author. Tel.: +86 538 8242301. E-mail address: [email protected] (Y. Liang).
1
25
These authors contributed equally to this work.
30
1
Page 1 of 21
JIAO J et al
J Plant Physiol
SUMMARY Fusaric acid (FA), a non-specific toxin produced mainly by Fusarium spp., can 35
cause programmed cell death (PCD) in tobacco suspension cells. The mechanism underlying the FA-induced PCD was not well understood. In this study, we analyzed the roles of hydrogen peroxide (H2O2) and mitochondrial function in the FA-induced PCD. Tobacco suspension cells
ip t
were treated with 100 μM FA and then analyzed for H2O2 accumulation and mitochondrial
functions. Here we demonstrate that cells undergoing FA-induced PCD exhibited H2O2 production, lipid peroxidation, and a decrease of the catalase and ascorbate peroxidase activities. Pre-treatment
cr
40
of tobacco suspension cells with antioxidant ascorbic acid and NADPH oxidase inhibitor diphenyl
us
iodonium significantly reduced the rate of FA-induced cell death as well as the caspase-3-like protease activity. Moreover, FA treatment of tobacco cells decreased the mitochondrial membrane
45
an
potential and ATP content. Oligomycin and cyclosporine A, inhibitors of the mitochondrial ATP synthase and the mitochondrial permeability transition pore, respectively, could also reduce the
M
rate of FA-induced cell death significantly. Taken together, the results presented in this paper demonstrate that H2O2 accumulation and mitochondrial dysfunction are the crucial events during
Keywords: Fusaric acid
Ac ce p
Programmed cell death
te
50
d
the FA-induced PCD in tobacco suspension cells.
Hydrogen peroxide Mitochondrion
55
Tobacco suspension cells Introduction
Programmed cell death (PCD) is an active process that regulates plant organ development and
cellular homeostasis (Green and Reed, 1998). Previous studies have indicated that during plant 60
growth and development, PCD can be induced by multiple biotic and abiotic stress stimuli including salt stress and toxins produced by plant pathogens (Yao et al., 2001; Duval et al., 2005; Lin et al., 2006; Samadi and Behboodi 2006; Wang et al., 2010; Jiao et al., 2013). A number of studies have also indicated that mitochondria, chloroplast, and signalling molecules have profound effects on PCD (Balk and Leaver, 2001; Vacca et al., 2006; Gadjev et al., 2008; Reape and 2
Page 2 of 21
JIAO J et al
65
J Plant Physiol
McCabe, 2010; Godbole et al., 2013). Fusaric acid (FA, 5-butylpicolinic acid) is a non-specific toxin produced by Fusarium species (Bacon et al., 1996), a common phytopathogen of many economically important plant species. Although the role of FA in Fusarium pathogenicity has not been fully characterized (Gapillout et
70
ip t
al., 1996), it is known to function as a virulence factor and its production can affect disease
symptom development. Several studies have shown that higher concentrations of FA (>10–4 M)
cr
were toxic to plant and can cause cell growth, change membrane permeability, decrease of
mitochondrial activity, inhibition of ATP synthesis, and change antioxidant enzyme activities
us
(Marré et al., 1993; Kuźniak, 2001). In contrast, low concentration of FA (
S0176-1617(14)00087-X http://dx.doi.org/doi:10.1016/j.jplph.2014.03.015 JPLPH 51923
To appear in: Received date: Revised date: Accepted date:
18-10-2013 8-3-2014 19-3-2014
Please cite this article as: Jiao J, Sun L, Zhou B, Gao Z, Hao Y, Zhu X, Liang Y, Hydrogen peroxide production and mitochondrial dysfunction contribute to the fusaric acid-induced programmed cell death in tobacco cells, Journal of Plant Physiology (2014), http://dx.doi.org/10.1016/j.jplph.2014.03.015 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
JIAO J et al
J Plant Physiol
Hydrogen peroxide production and mitochondrial dysfunction contribute to the fusaric acid-induced programmed cell death in tobacco cells Jiao Jiaoa,1, Ling Suna,1, Benguo Zhoub,1, Zhengliang Gaob, Yu Haoa, Xiaoping Zhua, Yuancun
ip t
5
a
cr
Lianga*
Department of Plant Pathology,
10
b
us
Shandong Agricultural University, Taian 271018, Shandong Province, China Tobacco Research Institute,
an
Anhui Academy of Agricultural Sciences, Hefei 23003, Anhui Province, China
Abbreviations: APX, ascorbate peroxidase; AsA, ascorbic acid; CAT, catalase; CsA, cyclosporine
15
M
A; DHR123, dihydrohodamine123; DPI, diphenyl iodonium; FA, fusaric acid; H2O2, hydrogen peroxide; MDA, malondialdehyde; MPTP, mitochondrial permeability transition pore; NO, nitric
te
death.
d
oxide; PBS, phosphate buffered saline; ROS, reactive oxygen species; PCD, programmed cell
20
Ac ce p
* Corresponding author. Tel.: +86 538 8242301. E-mail address: [email protected] (Y. Liang).
1
25
These authors contributed equally to this work.
30
1
Page 1 of 21
JIAO J et al
J Plant Physiol
SUMMARY Fusaric acid (FA), a non-specific toxin produced mainly by Fusarium spp., can 35
cause programmed cell death (PCD) in tobacco suspension cells. The mechanism underlying the FA-induced PCD was not well understood. In this study, we analyzed the roles of hydrogen peroxide (H2O2) and mitochondrial function in the FA-induced PCD. Tobacco suspension cells
ip t
were treated with 100 μM FA and then analyzed for H2O2 accumulation and mitochondrial
functions. Here we demonstrate that cells undergoing FA-induced PCD exhibited H2O2 production, lipid peroxidation, and a decrease of the catalase and ascorbate peroxidase activities. Pre-treatment
cr
40
of tobacco suspension cells with antioxidant ascorbic acid and NADPH oxidase inhibitor diphenyl
us
iodonium significantly reduced the rate of FA-induced cell death as well as the caspase-3-like protease activity. Moreover, FA treatment of tobacco cells decreased the mitochondrial membrane
45
an
potential and ATP content. Oligomycin and cyclosporine A, inhibitors of the mitochondrial ATP synthase and the mitochondrial permeability transition pore, respectively, could also reduce the
M
rate of FA-induced cell death significantly. Taken together, the results presented in this paper demonstrate that H2O2 accumulation and mitochondrial dysfunction are the crucial events during
Keywords: Fusaric acid
Ac ce p
Programmed cell death
te
50
d
the FA-induced PCD in tobacco suspension cells.
Hydrogen peroxide Mitochondrion
55
Tobacco suspension cells Introduction
Programmed cell death (PCD) is an active process that regulates plant organ development and
cellular homeostasis (Green and Reed, 1998). Previous studies have indicated that during plant 60
growth and development, PCD can be induced by multiple biotic and abiotic stress stimuli including salt stress and toxins produced by plant pathogens (Yao et al., 2001; Duval et al., 2005; Lin et al., 2006; Samadi and Behboodi 2006; Wang et al., 2010; Jiao et al., 2013). A number of studies have also indicated that mitochondria, chloroplast, and signalling molecules have profound effects on PCD (Balk and Leaver, 2001; Vacca et al., 2006; Gadjev et al., 2008; Reape and 2
Page 2 of 21
JIAO J et al
65
J Plant Physiol
McCabe, 2010; Godbole et al., 2013). Fusaric acid (FA, 5-butylpicolinic acid) is a non-specific toxin produced by Fusarium species (Bacon et al., 1996), a common phytopathogen of many economically important plant species. Although the role of FA in Fusarium pathogenicity has not been fully characterized (Gapillout et
70
ip t
al., 1996), it is known to function as a virulence factor and its production can affect disease
symptom development. Several studies have shown that higher concentrations of FA (>10–4 M)
cr
were toxic to plant and can cause cell growth, change membrane permeability, decrease of
mitochondrial activity, inhibition of ATP synthesis, and change antioxidant enzyme activities
us
(Marré et al., 1993; Kuźniak, 2001). In contrast, low concentration of FA (
