"Reactive Oxygen Species (ROS) and Reactive Nitrogen Species (NOS) in plant-pathogen interaction.

What do we know for a plant-nematode interaction?

PRIVILEGE TUNGAMIRAI MAKUNDE

PHYSIOLOGY OF PLANTS UNDER BIOTIC STRESS

Presentation layout
What is ROS and NRS?

Mechanisms of ROS and NRS Production in response to Pathogens

Roles of ROS and NRS following infection by plant pathogens

Interaction of ROS , NRS with other plant defence signal

ROS and NRS in plant-nematode interaction

What is ROS and NRS?

Reactive Oxygen Species (ROS)

By-products constantly produced during normal metabolic processes, such as photosynthesis or glycolysis resulting from one electron reduction from of molecular oxygen
early stages of plant-microorganism interactions Radicals:

O2

.-

Superoxide
Hydroxyl
.

Non-Radicals:

.OH

H2O2 HOCl1O 2

Hydrogen peroxide Hypochlorous acid

RO2 Peroxyl RO. Alkoxyl HO2 Hydroperoxyl

.

Singlet oxygen
Peroxynitrite

ONOO-

What is ROS and NRS?

Reactive Nitrogen Species (RNS)

Radicals: NO. Nitric Oxide NO2. Nitrogen dioxide

Non-Radicals: ONOOPeroxynitrite RSNOs S nitrosothiols GSNO S -nitrosoglutathione ROONO Alkyl peroxynitrites N2O3 Dinitrogen trioxide N2O4 Dinitrogen tetroxide HNO2 Nitrous acid NO2+ Nitronium anion NONitroxyl anion NO+ Nitrosyl cation NO2Cl Nitryl chloride

Mechanisms of ROS and NRS Production in response to Pathogens

Attempted infection by avirulent pathogens elicits a battery of plant defence system: elicit a biphasic ROS and NRS accumulation with a low-amplitude, transient first phase Preceding the 1st phase, a sustained phase of much higher magnitude that correlates with disease resistance is produced!! Leading to HYPERSENSITIVE RESPONSE ...a form of programmed cell death that contributes to plant resistance by restricting invading pathogens at the infection site virulent pathogens ??

Mechanisms of ROS and NRS Production in response to Pathogens

Several enzymes are implicated in apoplastic ROS production following successful pathogen recognition.
The main ROS producing enzymes are: plasma-membrane-bound NADPH oxidases

cell-wall-bound peroxidases
amine oxidases in the apoplast Superoxide anion (O2-) hydrogen peroxide (H2O2) hydroxyl radical Proof! NADPH oxidases (inhibited by diphenylene iodonium [DPI]) cell wall peroxidases (inhibited bycyanide or azide)( Grant et al., 2000a)

Mechanisms of ROS and NRS Production in response to Pathogens

Three routes are known to yield NO in plants reduction in nitrite by nitrate reductase (NR) oxidation of Arg to citrulline by NO synthase (NOS) a non-enzymatic NO generation system ROS and NRS molecules are highly reactive and toxic lead to the oxidative destruction of cells (Asada and Takahashi, 1987).

Mechanisms of ROS and NRS Production in response to Pathogens

Hirofumi et al ., 2010

NO and ROS are produced simultaneously through the MAPK cascade MEK2-SIPK

Roles of ROS and NRS following infection by plant pathogens

they are directly toxic to invading microorganisms ROS and NRS orchestrates the establishment of plant defence response, through defence gene activation for disease resistance and HR

provoke damage to DNA and proteins

regulate the synthesis of new signals such as salicylic acid ROS,NO in concert with SA, mediates the establishment of systemic defences (systemic acquired resistance [SAR]) The rapid production of ROS and the potential for H2O2 to freely diffuse across membranes suggested that; ROS could function as an intercellular or intracellular second messenger

Roles of ROS and NRS following infection by plant pathogens

ROS production has been associated with the formation of defensive barriers against powdery mildew in barley (Hordeum vulgare) strengthening of host cell walls cross-linking of glycoproteins

or lipid peroxidation membrane damage

Torres et al 2006

Roles of ROS and NRS following infection by plant pathogens

Gara et al 2003

Roles of ROS and NRS following infection by plant pathogens

COMPATIBLE INTERACTION

some pathogens may induce production of ROS and NRS to their own advantage exemplar
necrotrophs stimulates ROS and

NRS production in the infected tissue to induce cell death that facilitates subsequent infection There are also reports of ROS being produced, together with increased levels of ROS detoxification enzymes, during compatible interactions involving virus (Allan et al., 2001; Clarke et al., 2002).

Interaction of ROS and NRS with other plant defence signal

ROS is proposed to act synergistically in a signal amplification loop with SA to drive the HR and the establishment of systemic defences

SA accumulation can also down-regulate the ROS scavenging systems, in turn, contributes to increased overall ROS levels following pathogen recognition
Interestingly, both ROS and NO collaborate to mediate abscisic acid (ABA)-induced stomata closure (Desikan et al., 2004). Collectively, the interplay between these molecules mediates a variety of physiological responses

Interaction of ROS and NRS with other plant defence signal

Both ROS and ethylene have been implicated in signalling in response to viral infection (Love et al., 2005). Interestingly, the ethylene receptor ETR1 can function as an ROS sensor, mediating stomatal closure in response to H2O2 (Desikan et al., 2005). This cross talk may account for the multiplicity of responses mediated by ROS and explain why ROS produced by the same mechanism exert variable effects in different contexts.

Plant-Pathogen Interaction
Effects of silencing NbRibA on nitric oxide (NO) and oxidative bursts

[NO]

Signal intensity

Detection of ROS

Chemiluminescence

Asai et al 2010

Plant-Pathogen Interaction
Effects of silencing NbRibA on nitric oxide (NO) and oxidative bursts

biomass of P. infestans

10 FOLDS S
number of lesion spots on inoculated leaves

diameter of the lesions formed Asai et al 2010

ROS and NRS plant-nematode interaction

Increased peroxidase activity is also reported after nematode invasion produce bursts of H2O2 that have local effects on both nematodes and plant cells The latter may include: intermolecular linkages in the wall matrix in wounding repair catalysis of phenoxyl radical formation for lignification polymerisation of phenolic molecules in the production of suberin polysaccharide gelling NO and ROS are key molecules which may help to orchestrate events following nematode challenge

ROS and NRS plant-nematode interaction

Two M. incognita isolates, avirulent and virulent Versus Resistant tomato Mi gene

NO---Monitoring DAF-2DA fluorescence

Melillo et al 2011

ROS and NRS plant-nematode interaction

Two M. incognita isolates, avirulent and virulent Versus Resistant tomato Mi gene
[O2]

[H2O2] ---Amplex Red H2O2/Peroxidase assay kit

Melillo et al 2011

ROS and NRS plant-nematode interaction

Therefore nematode invasion of roots in some case induces hypersensitivity leading to localized plant cell death.

Melillo et al 2011

NO and ROS have a number of complementary, synergistic, and overlapping functions in plants HR results from the simultaneous, balanced production of NO and ROS

References
Allan AC, Lapidot M, Culver JN, Fluhr R (2001) An early tobacco mosaic virusinduced oxidative burst in tobacco indicates extracellular perception of the virus coat protein. Plant Physiol 126: 97108 Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu. Rev. Plant Biol. 55, 373-399 Asada K, Takahashi M (1987) Production and scavenging of active oxygen in photosynthesis. In DJ Kyle, CB Osmond, CJ Amtzen, eds, Photoinhibition (Topics in Photosynthesis), Vol 9. Elsevier, Amsterdam, pp 227287 Asai S, Mase K and Yoshioka H (2010) A key enzyme for flavin synthesis is required for nitric oxide and reactive oxygen species production in disease resistance Grant JJ, Loake GJ (2000) Role of reactive oxygen intermediates and cognate redox signaling in disease resistance. Plant Physiol 124: 2129

References
Melillo M. T, Leonetti P, Leone .A ,Veronico P and Zacheo T.B (2011) ROS and NO production in compatible and incompatible tomato-Meloidogyne incognita interactions. Eur J Plant Pathol Shetty N.P,Hans J. L, Jens J, David B. C and H. S Shetty (2008) Roles of reactive oxygen species in interactions between plants and pathogens. Eur J Plant Pathol Torres, M.A, Jones J. D.G, and Dangl J.L (2006) Reactive Oxygen Species Signaling in Response to Pathogens Plant Physiology journal Vol. 141, pp. 373378,
Liu X, Williams C .E,Nemacheck J.A, Wang H, Subramanyam S, Zheng C, and Ming-Shun C (2010) Reactive Oxygen Species Are Involved in Plant Defense against a Gall Midge