Beruflich Dokumente
Kultur Dokumente
2005
28410121020
Original Article
Oxidative stress and ozone: perception, signalling and response
M. Baier
et al.
Biochemistry & Physiology of Plants, W5, University of Bielefeld, 33501 Bielefeld, Germany
ABSTRACT
The primary site of ozone interaction with plant cells is the
extracellular matrix where ozone challenges the antioxidant protection of the cells. Accordingly, ozone sensitivity
generally correlates with the ascorbate status of the apoplast, which is an important signal initiation point. In addition, ozone sensing takes place by covalent modification of
redox-sensitive components of the plasma membrane, for
example ion channels like the plasma membrane Ca2+channels. Subsequent intracellular signal transduction is an
intriguing network of hormone, Ca2+ and MAPK signalling
pathways, significantly overlapping with oxidative burstinduced pathogen signalling. Comparison of recent
transcriptome analysis revealed that in addition to genes
generally induced by all kinds of oxidative stress, for example, transcripts for PR-proteins and most antioxidant
enzymes, approximately one-third of the responsive transcripts are ozone specific, indicating jasmonic acid, salicylic
acid and ethylene-independent redox signalling triggered
by extracellular redox sensing.
Key-words: abiotic stress; oxidative stress; ozone; reactive
oxygen species; redox signalling; signal transduction;
transcriptomics.
Abbreviations: 2CPA, 2-Cys peroxiredoxin-A; ABA,
abscisic acid; ACC, 1-aminocyclopropane-1-carboxylic
acid; GSH, reduced glutathione; JA, jasmonic acid; MAPK,
mitogen-activated protein kinase; RNS, reactive nitrogen
species; ROS, reactive oxygen species; SA, salicylic acid;
UV, ultra violet.
INTRODUCTION
Plants face a continuously changing environment. In order
to complete their life cycle and maintain a sufficient level
of fitness to reproduce, they need to adapt to abiotic and
biotic conditions. Periods of deviation from optimum are
sensed in order to anticipate severe stress and to activate
hardening processes. Two basic mechanisms allow for sensing of such environmental cues. First, sensors and receptors
have evolved to directly monitor physical or chemical
Correspondence, Margarete Baier.
E-mail: margarete.baier@uni-bielefeld.de
1012
ozone (Noctor & Foyer 1998; Plchl et al. 2000). Accordingly, when for instance in an ozone-sensitive genotype of
Raphanus sativus L. the L-ascorbate content was increased
by supplying the biosynthetic precursor L-galactono-1,4lactone, tolerance to ozone was increased (Maddison et al.
2002). However, using quite high concentrations of
300 p.p.b. O3, Luwe, Takahama & Heber (1993) observed a
very intriguing time-dependent relationship between oxidation of extracellular ascorbate and subsequent oxidation
of the intracellular glutathione pool, while the cellular
ascorbate redox state was unaltered during fumigation.
Ascorbate regeneration was tightly coupled to GSH within
the cell and transport activity was indicated to replenish the
reduced ascorbate pool in the apoplast. By the glutathionedependent regeneration mechanism, the extracellular
ascorbate oxidation affected the intracellular thiol
signature.
Experimental work strongly indicates that ascorbate is
not the only signal initiation point in ozone sensing. As
demonstrated by Jacob & Heber (1998) using oxidationsensitive fluorescence dyes, even co-infiltration of leaves
with 10 mM ascorbic acid did not prevent ascorbic-independent oxidation reactions. The reactivity of ozone is too high
to be selective. From recent work several potential pathways of redox-dependent signalling can be deduced
(Fig. 1): Using aequorin as an intracellular Ca2+-indicator
Clayton et al. (1999) showed a specific increase in the intracellular Ca2+-concentration if atmospheric ozone reached
concentrations above 70 p.p.b. It indicates oxidative activation of Ca2+ channels similar to the response to ABAinduced H2O2 (Kwak et al. 2003; Suhita et al. 2004; Dietrich
et al. 2004). In accordance with this hypothesis, short-term
exposure to O3 decreased the stomata aperture (McAinsh
et al. 2002). In terms of signalling, the Ca2+ channel would
2+
Ca
?
Lipid
lipid signalling/JA
LOOH
O3
H2O2
H2O2
5
ascorbate
ascorbate
GSSG
dehydroascorbate
dehydroascorbate
GSH
O3
intercellular
gas space
cell
wall
plasma
membrane
ROS
cytosol
2005 Blackwell Publishing Ltd, Plant, Cell and Environment, 28, 10121020
Oxidative burst
Recent studies presented evidence that ozone activates an
oxidative burst and may induce a hypersensitive response
(HR) similar to defence responses of plants to pathogen
infections (see Kangasjrvi et al. 2005 for details). Involvement of the plasmamembrane NADPH oxidase in ozonetriggered ROS accumulation has been, for example, shown
in the Arabidopsis thaliana mutant rcd1 (radical-induced
cell death 1; Overmyer et al. 2000). In this work, application
of the NADPH oxidase inhibitor diphenylene iodonium
inhibited ROS accumulation and reduced leaf damage of
the rcd1 mutant indicating that ozone initiates active cellular ROS production (Overmyer et al. 2000). In addition, in
ROS
Cytosol
C a 2+
Nucleus
MAPK
Ethylene
SA
JA
PP2C
2005 Blackwell Publishing Ltd, Plant, Cell and Environment, 28, 10121020
ROS
JA and SA
ABA
PP2C
MKK1
Ethylene
MPK4
Response
A
MKK4/5
MPK6
MPK3
Response
B
MEK1/2
Response
C
2005 Blackwell Publishing Ltd, Plant, Cell and Environment, 28, 10121020
O3 (200 nL L-1) and comprehensively compared the signalling pathways of ethylene, JA and SA on ozone-responsive
gene expression. Approximately 75% of the ozoneresponsive transcripts were induced and 48 of 205 genes
were suppressed by O3. Among the 109 transcripts with
known functions, 33 are involved in metabolism like monodehydroascorbate reductase, glutaredoxin and pyruvate
kinase, 24 in cellular organization and biogenesis, 25 in cell
rescue/defence (e.g. glutathione S-transferase, PR4, Lascorbate peroxidase), 11 in signal transduction like cyclophilin ROC7, six in energy (e.g. chlorophyll a/b-binding
protein and the gamma subunit of the mitochondrial F1ATPase), five in protein synthesis and degradation, three
in transcription and two in transport. This study corresponds
to results of previous studies of genes for which activation
or suppression by O3 has been described. In response to O3
exposure, transcript levels of cytosolic ascorbate peroxidase
(Willekens et al. 1994; Kubo et al. 1995), lipoxygenase
(Maccarrone, Veldink & Vliegenhardt 1992), blue copperbinding protein (Langebartels et al. 2000), glutathione Stransferase, PR proteins and catalases (Lim, Woo & Nam
2003) were elevated, whereas transcript levels of chlorophyll a/b-binding protein (Miller, Arteca & Pell 1999), rbcS
(Glick et al. 1995; Miller et al. 1999), and Cu/Zn superoxide
dismutase (Kliebenstein, Monde & Last 1998) were
reduced. Additionally, elevated transcript levels of enzymes
such as glutathione S-transferase, glutaredoxin, cyclophilin
ROC7 and ascorbate peroxidase indicate the involvement
of thiol-/disulphide transitions and thiol-dependent redox
regulation in acclimation to O3 exposure. With a proteomics
approach Agrawal et al. (2002c) studied the effects of ozone
exposure in rice. The ozone treatment resulted, for example, in decreased accumulation of photosynthetic proteins
whereas induced accumulation was, for example, observed
for an ascorbate peroxidase, superoxide dismutase, and
pathogenesis-related (PR) proteins. By contrast to acute
treatments, chronic exposure to ozone in the field caused a
smaller proportion of the Arabidopsis thaliana transcriptome to be altered, with five times more genes downregulated than up-regulated (Miyazaki et al. 2004).
As exposure to high levels of O3 induces the synthesis of
ethylene, JA and SA and activates the corresponding signalling pathways (Sharma & Davis 1997; Rao et al. 2000a,
b; Kangasjrvi et al. 2005), the regulation of 157 O3-induced
genes was analysed in signal transduction deficient mutants
as the ethylene-insensitive ein2, JA-resistant jar1 and SAinsensitive npr1. While approximately 50% of the transcripts were inhibited in ein2 and jar1, only 20% of the
transcripts were inhibited in npr1 (Tamaoki et al. 2003a),
indicating that JA and ethylene signalling are more crucial
for the induction than SA signalling at least under these
particular conditions. Of the O3-induced genes with function in cell rescue and defence such as L-ascorbate peroxidase, glutathione S-transferase and hevein-like protein
(PR4) about 70% were controlled by ethylene and JA and
were suppressed by SA, suggesting that ethylene and JA
signalling are important for the induction of defence gene
expression as outlined above.
2005 Blackwell Publishing Ltd, Plant, Cell and Environment, 28, 10121020
OUTLOOK
Recent analysis of transcriptome patterns in wild-type
plants and mutants has expanded our knowledge about
responses that are induced by oxidative stress. However, in
contrast to the experimental approaches, where, for example, the ozone concentration was suddenly and strongly
increased, in nature, during a vegetation period, the maximum daily ozone concentrations peak several times a
month over periods of weeks and induce hardening of the
plants (Luwe 1996). Therefore, one can assume that most
responses described in short-term fumigation experiments
are acute stress reactions. To understand oxidative signalling in general, however, we will have to distinguish the
intracellular oxidative sensing mechanisms from the com-
ACKNOWLEDGMENTS
The authors work that is summarized in this review was
supported by the Deutsche Forschungsgemeinschaft
(FOR387, Di346/6; Ba 2011/2)
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2005 Blackwell Publishing Ltd, Plant, Cell and Environment, 28, 10121020