Hindawi Publishing Corporation

Journal of Botany
Volume 2012, Article ID 217037, 26 pages
doi:10.1155/2012/217037

Review Article
Reactive Oxygen Species, Oxidative Damage, and Antioxidative
Defense Mechanism in Plants under Stressful Conditions

Pallavi Sharma,1 Ambuj Bhushan Jha,2 Rama Shanker Dubey,1 and Mohammad Pessarakli3
1 Department of Biochemistry, Faculty of Science, Banaras Hindu University, Varanasi 221005, India
2 Crop Development Centre, Department of Plant Sciences, College of Agriculture and Bioresources, University of Saskatchewan,
51 Campus Drive, Saskatoon SK, Canada SK S7N 5A8
3
School of Plant Sciences, The University of Arizona, Forbes Building, Room 303, P.O. Box 210036, Tucson, AZ 85721-0036, USA

Correspondence should be addressed to Mohammad Pessarakli, pessarak@email.arizona.edu

Received 6 August 2011; Revised 12 February 2012; Accepted 12 February 2012

Academic Editor: Andrea Polle

Copyright © 2012 Pallavi Sharma et al. This is an open access article distributed under the Creative Commons Attribution License,
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Reactive oxygen species (ROS) are produced as a normal product of plant cellular metabolism. Various environmental stresses
lead to excessive production of ROS causing progressive oxidative damage and ultimately cell death. Despite their destructive
activity, they are well-described second messengers in a variety of cellular processes, including conferment of tolerance to various
environmental stresses. Whether ROS would serve as signaling molecules or could cause oxidative damage to the tissues depends
on the delicate equilibrium between ROS production, and their scavenging. Efficient scavenging of ROS produced during various
environmental stresses requires the action of several nonenzymatic as well as enzymatic antioxidants present in the tissues. In this
paper, we describe the generation, sites of production and role of ROS as messenger molecules as well as inducers of oxidative
damage. Further, the antioxidative defense mechanisms operating in the cells for scavenging of ROS overproduced under various
stressful conditions of the environment have been discussed in detail.

1. Introduction cell is said to be in a state of “oxidative stress.” The enhanced

production of ROS during environmental stresses can pose
An unavoidable consequence of aerobic metabolism is a threat to cells by causing peroxidation of lipids, oxidation
production of reactive oxygen species (ROS). ROS include of proteins, damage to nucleic acids, enzyme inhibition, acti-
free radicals such as superoxide anion (O2 •− ), hydroxyl vation of programmed cell death (PCD) pathway and ulti-
radical (• OH), as well as nonradical molecules like hydro- mately leading to death of the cells [6–8, 11, 13, 14, 16, 17].
gen peroxide (H2 O2 ), singlet oxygen (1 O2 ), and so forth. Despite their destructive activity, ROS are well-described
Stepwise reduction of molecular oxygen (O2 ) by high-energy second messengers in a variety of cellular processes including
exposure or electron-transfer reactions leads to production tolerance to environmental stresses [18–20]. Whether ROS
of the highly reactive ROS. In plants, ROS are always formed will act as damaging or signaling molecule depends on the
by the inevitable leakage of electrons onto O2 from the elec- delicate equilibrium between ROS production and scaveng-
tron transport activities of chloroplasts, mitochondria, and ing. Because of the multifunctional roles of ROS, it is neces-
plasma membranes or as a byproduct of various metabolic sary for the cells to control the level of ROS tightly to avoid
pathways localized in different cellular compartments [1– any oxidative injury and not to eliminate them completely.
5]. Environmental stresses such as drought, salinity, chilling, Scavenging or detoxification of excess ROS is achieved by
metal toxicity, and UV-B radiation as well as pathogens an efficient antioxidative system comprising of the nonen-
attack lead to enhanced generation of ROS in plants due zymic as well as enzymic antioxidants [21]. The enzymic
to disruption of cellular homeostasis [6–15]. All ROS are antioxidants include superoxide dismutase (SOD), catalase
extremely harmful to organisms at high concentrations. (CAT), guaiacol peroxidase (GPX), enzymes of ascorbate-
When the level of ROS exceeds the defense mechanisms, a glutahione (AsA-GSH) cycle such as ascorbate peroxidase
2 Journal of Botany

(APX), monodehydroascorbate reductase (MDHAR), dehy- dissipation during photosynthesis can lead to formation of
droascorbate reductase (DHAR), and glutathione reductase chlorophyll (Chl) triplet state, whereas in the reaction centre
(GR) [21]. Ascorbate (AsA), glutathione (GSH), carotenoids, it is formed via charge recombination of the light-induced
tocopherols, and phenolics serve as potent nonenzymic charge pair [29]. The Chl triplet state can react with 3 O2 to
antioxidants within the cell. Various workers have reported give up the very highly destructive ROS 1 O2 :
increased activities of many enzymes of the antioxidant
defense system in plants to combat oxidative stress induced light
Chl −−→ 3 Chl, (1)
by various environmental stresses. Maintenance of a high
antioxidant capacity to scavenge the toxic ROS has been 3
Chl + 3 O2 −→ Chl + 1 O2 , (2)
linked to increased tolerance of plants to these environmental
stresses [22, 23]. Considerable progress has been made in Further, limited CO2 availability due to closure of stomata
improving stress-induced oxidative stress tolerance in crop during various environmental stresses such as salinity,
plants by developing transgenic lines with altered levels of drought favors the formation of 1 O2 . The life time of 1 O2
antioxidants [24, 25]. Simultaneous expression of multiple within the cell is probably 3 µs or less [30, 31]. A fraction of
antioxidant enzymes has been shown to be more effective 1 O has been shown to be able to diffuse over considerable
2
than single or double expression for developing transgenic distances of several hundred nanometers (nm). 1 O2 can last
plants with enhanced tolerance to multiple environmental for 4 µs in water and 100 µs in a nonpolar environment
stresses [26]. The present review focuses on types of ROS, [1]. 1 O2 reacts with most of the biological molecules
their site of production, and their role as messenger and at near diffusion-controlled rates [1]. It directly oxidizes
inducer of oxidative stress. Further, role of antioxidative protein, unsaturated fatty acids, and DNA [32]. It causes
defense system in combating danger posed by overproduced nucleic acid modification through selective reaction with
ROS under stresses has been discussed in detail. deoxyguanosine [33]. It is thought to be the most important
species responsible for light-induced loss of photosystem II
(PSII) activity which may trigger cell death [34]. 1 O2 can be
2. Reactive Oxygen Species, Sites of quenched by β-carotene, α-tocopherol or can react with the
Production, and Their Effects D1 protein of photosystem II as target [29].
Due to spin restriction, molecular O2 cannot accept four
ROS are a group of free radicals, reactive molecules, and ions
electrons at a time to produce H2 O. It accepts one electron at
that are derived from O2 . It has been estimated that about 1%
a time and hence during reduction of O2 stable intermediates
of O2 consumed by plants is diverted to produce ROS [27] in
are formed in the step-wise fashion [35]. O2 •− is the
various subcellular loci such as chloroplasts, mitochondria,
primary ROS formed in the cell which initiates a cascade
peroxisomes. ROS are well recognized for playing a dual role
of reactions to generate “secondary” ROS, either directly or
as both deleterious and beneficial species depending on their
prevalently through enzyme- or metal-catalysed processes
concentration in plants. At high concentration ROS cause
[36] depending on the cell type or cellular compartment.
damage to biomolecules, whereas at low/moderate concen-
O2 •− is a moderately reactive, short-lived ROS with a half-
tration it acts as second messenger in intracellular signaling
life of approx. 1 µs. O2 •− is a nucleophilic reactant with
cascades that mediate several responses in plant cells.
both oxidizing and reducing properties [37]. Anionic charge
of O2 •− inhibits its electrophilic activity toward electron-
2.1. Types of ROS. The most common ROS include 1 O2 , rich molecules. O2 •− has been shown to oxidize enzymes
O2 •− , H2 O2 , • OH. O2 itself is a totally harmless molecule as containing the [4Fe-4S] clusters (aconitase or dehydratase
in its ground state it has two unpaired electrons with parallel as examples) [38] and reduce cytochrome C [39]. O2 •− can
spin which makes it paramagnetic and, hence, unlikely accept one electron and two protons to form H2 O2 . It is
to participate in reactions with organic molecules unless easily dismutated to H2 O2 either nonenzymatically or by
it is activated [28]. Activation of O2 may occur by two SOD catalyzed reaction to hydrogen peroxide:
different mechanisms: (i) absorption of sufficient energy to
reverse the spin on one of the unpaired electrons and (ii) 2O2 •− + 2H+ −→ H2 O2 + O2 , (3)
stepwise monovalent reduction (Figure 1). In the former,
1 O is formed, whereas in latter, O is sequentially reduced SOD
2 2 2O2 •− + 2H+ −−→ H2 O2 + O2 , (4)
to O2 •− , H2 O2 , and • OH (Figure 1).
Electrons in the biradical form of oxygen have parallel H2 O2 is generated in the cells under normal as well as
spin. Absorption of sufficient energy reverses the spin of one wide range of stressful conditions such as drought, chilling,
of its unpaired electrons leading to formation of singlet state UV irradiation, exposure to intense light, wounding and
in which the two electrons have opposite spin. This activation intrusion by pathogens. Electron transport chain (ETC)
overcomes the spin restriction and 1 O2 can consequently of chloroplast, mitochondria, endoplasmic reticulum and
participate in reactions involving the simultaneous transfer plasma membrane, β-oxidation of fatty acid and photorespi-
of two electrons (divalent reduction) [28]. In the light, highly ration are major sources of H2 O2 generation in plant cells.
reactive 1 O2 can be produced via triplet chlorophyll (Chl) Photooxidation reactions, NADPH oxidase as well as xan-
formation in the antenna system and in the reaction centre thine oxidase (XOD) also contribute to H2 O2 production in
of photosystem II [29]. In the antenna, insufficient energy plants. It is also generated in tissues requiring it as a substrate
Journal of Botany 3

1O
2 H2 O
Singlet Water
oxygen

CAT
e−
e- GPX
Energy absorption
APX
H+
Spontaneous
dismutation H2 O2
O2 O2 •−
e− e− 2H+ Hydrogen
Molecular Superoxide
oxygen anion peroxide
SOD e−

Fenton H+ Haber-
reaction Weiss
reaction

• OH

Hydroxyl
radical

Figure 1: Schematic representation of generation of reactive oxygen species (ROS) in plants. Activation of O2 occurs by two different
mechanisms. Stepwise monovalent reduction of O2 leads to formation of O2 •− , H2 O2 , and • OH, whereas energy transfer to O2 leads to
formation of 1 O2 . O2 •− is easily dismutated to H2 O2 either nonenzymatically or by superoxide dismutase (SOD) catalyzed reaction to H2 O2 .
H2 O2 is converted to H2 O by catalase (CAT), guaiacol peroxidase (GPX), and ascorbate peroxidase (APX).

for biosynthesis such as for lignification and suberization. First, Fe(III) is reduced by O2 •− , followed by oxidation by
H2 O2 is moderately reactive and is relatively long-lived dihydrogen peroxide (Fenton reaction)
molecule with a half-life of 1 ms [40]. H2 O2 has no unpaired
electrons, unlike other oxygen radicals, it can readily cross Fe2+ + H2 O2 −→ Fe3+ + OH− +• OH, (6)
biological membranes and consequently can cause oxidative
damage far from the site of its formation. Because H2 O2 is and reaction:
the only ROS that can diffuse through aquaporins in the
membranes and over larger distances within the cell [41] and O2 •− + H2 O2 −→ • OH + OH− + O2 . (7)
is relatively stable compared to other ROS, it has received
particular attention as a signal molecule involved in the
Metal catalysis is necessary for this reaction since the rate
regulation of specific biological processes and triggering tol-
of uncatalyzed reaction is negligible [47]. • OH is the most
erance against various environmental stresses such as plant-
reactive among all ROS. It has a single unpaired electron,
pathogen interactions at low concentration [19, 20, 42]. At
thus, it can react with oxygen in triplet ground state. • OH
high concentration, H2 O2 can oxidize the cysteine (–SH)
interacts with all biological molecules and causes subsequent
or methionine residues (–SCH3 ), and inactivate enzymes by
cellular damages such as lipid peroxidation, protein damage,
oxidizing their thiol groups, such as enzymes of Calvin cycle,
and membrane destruction [48]. Because cells have no enzy-
Cu/Zn-SOD, and Fe-SOD [43]. When hydrogen peroxide
matic mechanism to eliminate • OH, its excess production
accumulates at levels of 10 µM, the enzymes in the Calvin
can eventually lead to cell death [49]. Under illumination,
cycle, such as fructose-1,6-bisphosphatase, sedoheptulose-
formation of • OH by the Fenton reaction at the active site
1,7-bisphosphatase, and phosphoribulokinase, lose 50% of
of the enzyme RbcL leads to its fragmentation in chloroplast
their activity [44, 45]. It also oxidizes protein kinases,
lysates [50, 51]. The oxidation of organic substrates by • OH
phosphatases, and transcription factors containing thiolate
may proceed by two possible reactions, either by addition
residues. At high concentrations, it orchestrates programmed
of • OH to organic molecules or due to abstraction of a
cell death [46].
hydrogen atom from it. Because of short lifetime and the
Both O 2 •− and H2 O2 are only moderately reactive.
strongly positive redox potential (close to +2 V) of “free”
The cellular damage by ROS appears to be due to their •
OH, its sites of reaction are close to its point of formation
conversion into more reactive species. The formation of • OH
[52]. In this context, organic oxygen radicals such as alkoxy,
is dependent on both H2 O2 and O2 •− and, thus, its formation
peroxy, semiquinones, reduced hydrogen peroxide, and
is subject to inhibition by both SOD and CAT.
hydrogen peroxide-electron donor complexes (crypto-OH),
The Haber-Weiss reaction generates • OH from H2 O2 and
as well as metallo-oxygen complexes, have been proposed
O2 •− . It consists of the following two reactions:
as the ultimately active species besides destructive free • OH
Fe3+ + O2 •− −→ Fe2+ + O2 , (5) [53].
4 Journal of Botany

Chloroplast
Mitochondria
PSI: electron transport chain
Fd, 2Fe-2S, and 4Fe-4S clusters Complex I: NADH dehydrogenase segment
PSII: electron transport chain Complex II: reverse electron flow to complex I
QA and QB Complex III: ubiquinone-cytochrome region
Enzymes
Chlorophyll pigments Aconitase, 1-galactono-γ
lactone, dehydrogenase (GAL)
Cell wall
Cell-wall-associated peroxidase Plasma membrane
ROS Electron transporting oxidoreductases
diamine oxidases
NADPH oxidase, quinone oxidase

Endoplasmic Peroxisome
reticulum
NAD(P)H-dependent Matrix: xanthine oxidase (XOD)
electron transport Apoplast Membrane: electron transport chain flavoprotein
involving Cyt P450 Cell-wall-associated NADH and Cyt b
oxalate oxidase Metabolic processes: glycolate oxidase,
Amine oxidases fatty acid oxidation, flavin oxidases,
disproportionation of O2 •−
radicals

Figure 2: Sites of production of reactive oxygen species (ROS) in plants. ROS are produced at several locations in the cell-like chloroplast,
mitochondria, plasma membrane, peroxisomes, apoplast, endoplasmic reticulum, and cell wall.

2.2. Sites of Production of ROS. ROS are produced in both spontaneously to H2 O2 on the external “stromal” membrane
unstressed and stressed cells at several locations in chloro- surface. At Fe-S centers where Fe2+ is available, H2 O2 may
plasts, mitochondria, plasma membranes, peroxisomes, be transformed through the Fenton reaction into the much
apoplast, endoplasmic reticulum, and cell walls (Figure 2). more dangerous OH• .
ROS are always formed by the inevitable leakage of electrons
onto O2 from the electron transport activities of chloroplasts,
mitochondria, and plasma membranes or as a byproduct 2.2.2. Mitochondria. Mitochondria can produce ROS in
of various metabolic pathways localized in different cellular several sites of ETC. In mitochondria direct reduction of
compartments. oxygen to O2 •− occurs in the flavoprotein region of NADH
dehydrogenase segment (complex I) of the respiratory chain
2.2.1. Chloroplasts. In chloroplasts, various forms of ROS [56]. When NAD+ -linked substrates for complex I are
are generated from several locations. ETCs in PSI and PSII limited, electron transport can occur from complex II to
are the main sources of ROS in chloroplasts. Production of complex I (reverse electron flow). This process has been
ROS by these sources is enhanced in plants by conditions shown to increase ROS production at complex I and is
limiting CO2 fixation, such as drought, salt, and temperature regulated by ATP hydrolysis [57]. Ubiquinone-cytochrome
stresses, as well as by the combination of these conditions region (complex III) of the ETC also produces O2 •− from
with high-light stress. Under normal conditions, the electron oxygen. It is believed that fully reduced ubiquinone donates
flow from the excited PS centers to NADP which is reduced an electron to cytochrome C1 and leaves an unstable highly
to NADPH which, then, enters the Calvin cycle and reduces reducing ubisemiquinone radical which is favorable for the
the final electron acceptor, CO2 . In case of overloading of the electron leakage to O2 and, hence, to O2 •− formation [58].
ETC, due to decreased NADP supply resulting from stress In plants, under normal aerobic conditions, ETC and ATP
conditions, there is leakage of electron from ferredoxin to syntheses are tightly coupled; however, various stress factors
O2 , reducing it to O2 •− [54]. This process is called Mehler lead to inhibition and modification of its component, leading
reaction: to over reduction of electron carriers and, hence, formation
of ROS [4, 59].
2O2 + 2Fdred −→ 2O2 •− + 2Fdox (8) Several enzymes present in mitochondrial matrix can
produce ROS. Some of them produce ROS directly, for
Leakage of electrons to O2 may also occur from 2Fe-2S and example aconitase, whereas some others like 1-galactono-
4Fe-4S clusters in the ETC of PSI. In PSII, acceptor side of γ lactone dehydrogenase (GAL), are able to feed electrons
ETC contains QA and QB. Leakage of electron from this site to ETC [60, 61]. O2 •− is the primary ROS formed by
to O2 contributes to the production of O2 •− [55]. monovalent reduction in the ETC. It is converted quickly
The formation of O2 •− by O2 reduction is a rate-limiting either by the MnSOD (mitochondrial form of SOD) or APX
step. Once formed O2 •− generates more aggressive ROS. into the relatively stable and membrane-permeable H2 O2 .
It may be protonated to HO2 • on the internal, “lumen” H2 O2 can be further converted to extremely active hydroxyl
membrane surface or dismutated enzymatically (by SOD) or radical (OH• ) in the Fenton reaction.
6 Journal of Botany

Mediated
Hormones Plant responses

Abscisic acid Stomata closure

Auxin Root gravitropism

Gibberelic acid Seed germination

Messenger
ROS Lignin biosynthesis
at low concentration Jasmonic acid

Gibberelic acid Programmed cell death

Salicylic acid Hypersensitive responses

Salicylic acid Osmotic stress

Figure 3: Reactive oxygen species (ROS) as second messengers in several plant hormone responses, including stomatal closure, root
gravitropism, seed germination, lignin biosynthesis, programmed cell death, hypersensitive responses, and osmotic stress.

[19]. The strength, lifetime and size of the ROS signaling and suggested that ROS are components of the hormonally
pool depends on balance between oxidant production and regulated cell death pathway in barley aleurone cells.
removal by the antioxidant. Using mutants deficient in Plants have evolved a complex regulatory network to
key ROS-scavenging enzymes, Miller and coworkers [74] mediate biotic and abiotic stress responses based on ROS
identified a signaling pathway that is activated in cells synthesis, scavenging, and signaling. Transient production of
in response to ROS accumulation. Interestingly, many of ROS is detected in the early events of plant-pathogen inter-
the key players in this pathway, including different zinc actions and plays an important signaling role in pathogenesis
finger proteins and WRKY transcription factors, are also signal transduction regulators. This production-called oxida-
central regulators of abiotic stress responses involved in tive burst could be considered as a specific signal during the
temperature, salinity and osmotic stresses. interaction process [80]. In HR, SA is thought to potentiate
ROS are considered second messengers in the abscisic ROS signaling [81]. ROS are shown to act as a second
acid (ABA) transduction pathway in guard cells [19, 20]. messenger for the induction of defense genes in tomato
ABA induced H2 O2 is an essential signal in mediating plants in response to wounding [82]. ROS were generated
stomatal closure to reduce water loss through the activation near cell walls of vascular bundle cells of tomato leaves
of calcium-permeable channels in the plasma membrane in response to wounding and resulted H2 O2 from wound-
[77]. Jannat and coworkers [78] observed that ABA- inducible polygalacturonase acted as a second messenger for
inducible cytosolic H2 O2 elevation functions in ABA- the activation of defense genes in mesophyll cells, but not for
induced stomatal closure, while constitutive increase of signaling pathway genes in vascular bundle cells [82].
H2 O2 does not cause stomatal closure. Role of ROS as second Lignin is important for the plant’s response to environ-
messenger in root gravitropism has been demonstrated. mental stress. Denness and coworkers [83] characterized a
Based on their work, Joo and coworkers [73] proposed genetic network enabling plants to regulate lignin biosynthe-
that gravity induces asymmetric movement of auxin within sis in response to cell wall damage through dynamic interac-
60 min, and, then, the auxin stimulates ROS generation tions between Jasmonic acid and ROS. ROS have been shown
to mediate gravitropism. Further, scavenging of ROS by to play important roles in osmotic stress, low temperature,
antioxidants (N-acetylcysteine, ascorbic acid, and Trolox) and heavy metal signal transduction pathway [75, 84, 85].
inhibited root gravitropism [73]. ROS appear to be involved Genes involved in osmotic stress signaling have been shown
in dormancy alleviation. In dormant barley grains under to be upregulated by ROS, including the transcription factor
control condition, gibberellic acid (GA) signaling and ROS DREB2A and a histidine kinase [18]. In Arabidopsis culture
content are low, while ABA signaling is high, resulting in cells, it was reported that the MAPK AtMPK6 that can be
dormancy. Exogenous H2 O2 does not appear to alter ABA activated by low temperature and osmotic stress could also
biosynthesis and signaling, but has a more pronounced effect be activated by oxidative stress [84]. Borsani and coworkers
on GA signaling, inducing a change in hormonal balance [86] suggested that the increased osmotic stress tolerance
that results in germination [79]. ROS have been shown to of transgenic Arabidopsis expressing a salicylate hydroxylase
play a key role in PCD in barley aleurone cells, initiated (NahG) gene, might result from decreased SA-mediated ROS
by GA. Bethke and Jones [72] observed that GA-treated generation. Zhao and coworkers [87] reported that ROS play
aleurone protoplasts are less tolerant to internally generated important roles in drought-induced abscisic acid synthesis
or exogenously applied H2 O than ABA-treated protoplasts in plant and suggested that they may be the signals through
Journal of Botany 7

ROS
at high concentrations

Oxidative damage

Lipid Protein DNA

Chain breakage Site-specific amino acid Deoxyribose oxidation
Increase in membrane modification Strand breakage
fluidity and permeability Fragmentation of the Removal of nucleotides
peptide chain Modification of bases
Aggregation of cross- DNA-protein crosslinks
linked reaction products
Altered electric charge
Enzyme inactivation
Increased susceptibility
of proteins to proteolysis

Figure 4: Reactive oxygen species (ROS) induced oxidative damage to lipids, proteins, and DNA.

which the plant can “sense” the drought condition. Using acid. The polyunsaturated fatty acids (PUFAs) present in
pharmacological inhibitors, it is demonstrated that metals membrane phospholipids are particularly sensitive to attack
Cd2+ and Cu2+ induce MAP kinase activation via distinct by ROS. A single • OH can result in peroxidation of many
ROS-generating systems [85]. polyunsaturated fatty acids because the reactions involved in
this process are part of a cyclic chain reaction. The overall
2.4. ROS and Oxidative Damage to Biomolecules. Production process of lipid peroxidation involves three distinct stages:
and removal of ROS must be strictly controlled in order to initiation, progression, and termination steps. The initial
avoid oxidative stress. When the level of ROS exceeds the phase of lipid peroxidation includes activation of O2 which is
defense mechanisms, a cell is said to be in a state of “oxidative rate limiting. O2 •− and • OH can react with methylene groups
stress”. However, the equilibrium between production and of PUFA forming conjugated dienes, lipid peroxy radicals
scavenging of ROS is perturbed under a number of stressful and hydroperoxides [88]:
conditions such as salinity, drought, high light, toxicity due PUFA − H + X• −→ PUFA + X − H. (9)
to metals, pathogens, and so forth. Enhanced level of ROS
can cause damage to biomolecules such as lipids, proteins
and DNA (Figure 4). These reactions can alter intrinsic PUFA + O2 −→ PUFA − OO• . (10)
membrane properties like fluidity, ion transport, loss of
The peroxy radical formed is highly reactive and able to
enzyme activity, protein cross-linking, inhibition of protein
propagate the chain reaction:
synthesis, DNA damage, and so forth ultimately resulting in
cell death. PUFA − OO• +PUFA − OOH −→ PUFA − OOH + PUFA•
(11)
2.4.1. Lipids. When ROS level reaches above threshold,
The formation of conjugated diene occurs when free radicals
enhanced lipid peroxidation takes place in both cellular and
attack the hydrogens of methylene groups separating double
organellar membranes, which, in turn, affect normal cellular
bonds and, thereby, rearrangement of the bonds occurs
functioning. Lipid peroxidation aggravates the oxidative
[89]. The lipid hydroperoxides produced (PUFA-OOH) can
stress through production of lipid-derived radicals that
undergo reductive cleavage by reduced metals, such as Fe2+ ,
themselves can react with and damage proteins and DNA.
according to the following reaction:
The level of lipid peroxidation has been widely used as an
indicator of ROS mediated damage to cell membranes under Fe2+ complex + PUFA − OOH
stressful conditions. Increased peroxidation (degradation) of (12)
−→ Fe3+ complex + PUFA − O•
lipids has been reported in plants growing under environ-
mental stresses [8, 10, 12, 13]. Increase in lipid peroxidation Several reactive species including: lipid alkoxyl radicals, alde-
under these stresses parallels with increased production of hydes (malonyldialdehyde, acrolein and crotonaldehyde),
ROS. Malondialdehyde (MDA) is one of the final products alkanes, lipid epoxides, and alcohols can be easily formed
of peroxidation of unsaturated fatty acids in phospholipids by the decomposition of lipid hydroperoxide [90]. The lipid
and is responsible for cell membrane damage [43]. Two alkoxy radical produced, (PUFA-O• ), can initiate additional
common sites of ROS attack on the phospholipid molecules chain reactions [91]:
are the unsaturated (double) bond between two carbon
atoms and the ester linkage between glycerol and the fatty PUFA − O• + PUFA − H −→ PUFA − OH + PUFA• (13)