Reactive Oxygen Species (ROS)

Reactive oxygen species (ROS)
Presented by
B. Tarun Reddy
Reactive Oxygen Species
• ROS a collective term that includes all reactive
forms of oxygen includes both radicals and
non radicals.
• Free radicals represent a class of highly
reactive intermediate chemical entities whose
reactivity is derived from the presence of
unpaired electron.
Free Radical
 ROS are well recognized for playing a dual role as both
deleterious and beneficial species depending on their
concentration in plants.
 They are formed as a natural by-product of the normal
metabolism and have important roles in cell signaling
and homeostasis.
 Higher level of ROS can attack virtually all
macromolecules, which results in serious damage to
cellular components .
Whereas at low concentration it acts as
messenger in intracellular signaling.
Under stress conditions, the balance between
the production and elimination of ROS is
disturbed in cellular components of plants.
ROS production and scavenging is thought to
be controlled by a large network of genes
termed the ROS gene network.
Sources of ROS
Glycolate cycle
Electron
transport peroxisomes
chain
Exogenous sources of ROS
Site of production
• 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 of various metabolic pathways
localized in different cellular compartments.
Chloroplast
 ETCs in PSI and PSII are the main sources of ROS
in chloroplasts.
 ROS are produced in conditions such as drought,
salt, and temperature stresses, as well as by the
combination of these conditions with high-light
intensity.
2 O2 + 2 Fd (red) ⟶ 2 O2•− + 2 Fd (oxi)
Mitochondria
In plants, under normal aerobic conditions
and ATP syntheses are tightly coupled;
however, various stress factors lead to
inhibition or modification leading to over
reduction of electron carriers which leads to
formation of ROS.
 Several enzymes present in mitochondrial
matrix can produce ROS.
Peroxisomes
Peroxisomes are probably the major sites of
H2O2 production.
The main metabolic processes responsible for
the generation of H2O2 in peroxisomes are the
photorespiration, the fatty acid β-oxidation
and flavin oxidases.
Plasma membrane
Electron transporting oxidoreductases are at
plasma membranes and lead to generation of
ROS at plasma membrane.
NADPH oxidase transfer of electrons from
cytoplasmic NADPH to O2 to form O2 •−.
NADPH oxidase has been proposed to play a
key role in the production and accumulation
of ROS in plants under stress conditions.
Apoplast
Cell-wall-located enzymes are responsible for
apoplastic ROS production .
The cell-wall-associated oxalate oxidase,
releases H2O2 and CO2 from oxalic acid.
Amine oxidase like enzymes may contribute to
defense responses occurring in the apoplast
following biotic stress, mainly through H2O2
production.
Oxidative damage to
Biomolecules
ROS
at high concentration
Membranes
 When ROS level reaches above threshold, peroxidation
takes place in which in turn affect normal cellular
functioning.
 A single •OH can result in peroxidation of many fatty
acids molecules.
 These ROS attack can lead to chain breakage and,
thereby, increase in membrane fluidity and
permeability.
Proteins
 Amino acid modification, fragmentation of the peptide
chain, aggregation of cross-linked reaction products,
altered electric charge and increased susceptibility of
proteins to proteolysis occur.
 Thiol groups and Sulphur containing amino acids are very
susceptible sites for attack by ROS.
 Oxidized proteins were more efficiently degraded by
proteolytic activity increased.
DNA
DNA is genetic material and any damage to the DNA
can result in changes in proteins, which may lead to
malfunctions or complete inactivation of the proteins.
Oxidative attack on DNA results in deoxyribose

oxidation, strand breakage, removal of nucleotides,
variety of modifications in the organic bases of the
nucleotides, and DNA-protein crosslinks.
When •OH attacks on either DNA or proteins
associated with it, DNA protein crosslinks are formed .
DNA protein crosslinks cannot be readily repaired and
may be lethal if replication or transcription precedes.
Mitochondrial and chloroplast DNA are more
susceptible to oxidative damage than nuclear DNA.
Antioxidant defense system in
plants
Super oxide dismutase (SOD)
• The enzyme sod belongs to the group of
metalloenzymes first discovered by McCord
and Fridovich (1969) .
• Three isozymes of SOD are
• Cu/Zn-SOD(cytosol, chloroplast, mitochondria
and peroxisomes).
• Mn-SOD(mitochondria)
• Fe-SOD(chloroplast).
Catalase (CAT)
• It is a ubiquitous tetrameric heme containing
enzyme.
• CAT scavenges H2O2 generated in this
organelle during photorespiratory oxidation,
β-oxidation of fatty acids coupled to SOD.
APX, MDHAR, DHAR, GR
Non enzymatic antioxidents
Tocopherols
• Tocopherols (α, β, γ, and δ) represent a group of lipophilic
antioxidants involved in scavenging of oxygen free radicals.
• Synthesized only by photosynthetic organisms.
• Regeneration of the oxidized tocopherol back to its reduced
form can be achieved by AsA, GSH.
• Accumulation of α-tocopherol has been shown to induce
tolerance to chilling, water deficit, and salinity in different
plant species.
Carotenoids
• Carotenoids are found in plants as well as
microorganism.
• As an antioxidant, they scavenge 1O2 (singlet
oxygen) to inhibit oxidative damage.
• Carotenoids also serve as precursors to signaling
molecules that influence plant development and
biotic/abiotic stress responses.
Oxidative stress
 The accumulation of reactive oxygen species
(ROS), which damage cellular structures is called
as oxidative stress.
 The equilibrium between production and
scavenging of ROS is disturbed under a number of
stressful conditions such as salinity, drought, high
light, toxicity due to metals, pathogens, and so
on.
 Enhanced level of ROS can cause damage to
biomolecules such as lipids, proteins and DNA .
• When pathogen attacks the defense reaction,
it leads to the production of ROS, primarily as
superoxide and H2O2, that finally leads to
various defense mechanisms against the
pathogens.
• Antimicrobial approaches primarily utilizing
ROS comprise both bactericidal antibiotics and
non- pharmacological methods such as
photodynamic therapy, titanium dioxide
photocatalysis, cold plasma, and medicinal
honey.
ROS in cell signaling
• The ROS signals are auto-propagating in
nature and transferred throughout over a long
distance in plants such as in Arabidopsis
thaliana wherein signal propagates at a rate of
8.4 cm/ min.
• Another key signaling advantage of ROS is
their tight link to cellular homeostasis and
metabolism.
Conclusion
 Study of formation and fate of ROS using advanced
analytical techniques will help in developing broader
view of the role of ROS in plants.
 Future progress in genomics, metabolomics, and
proteomics will help in clear understanding of
biochemical networks involved in cellular responses to
oxidative stress.
 Improved understanding of these will be helpful in
producing plants with in-built capacity of enhanced
levels of tolerance to ROS using biotechnological
approach.
Reference
• G. Tanou, A. Molassiotis, and G. Diamantidis, “Induction of
reactive oxygen species and necrotic death-like destruction
in strawberry leaves by salinity,” Environmental and
Experimental Botany, vol. 65, no. 2-3, pp. 270–281, 2009.
• L. A. Del R´ıo, L. M. Sandalio, F. J. Corpas, J. M. Palma, and J.
B. Barroso, “Reactive oxygen species and reactive nitrogen
species in peroxisomes. Production, scavenging, and role in
cell signaling,” Plant Physiology, vol. 141, no. 2, pp. 330–
335, 2006.
• Y. Yamauchi, A. Furutera, K. Seki, Y. Toyoda, K. Tanaka, and Y.
Sugimoto, “Malondialdehyde generated from peroxidized
linolenic acid causes protein modification in heat-stressed
plants,” Plant Physiology and Biochemistry, vol. 46, no. 8-9,
pp. 786–793, 2008.

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Oxidative attack on DNA results in deoxyribose

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