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Molecular events during plant-pathogen interactions

Compatible and incompatible interactions

As a consequence of the sessile existence, plants experience intense exposure of plethora of

pathogens. Plants have evolved multiple mechanisms to recognize pathogens and to induce
both local and systemic responses. The first line of defense is mediated by the recognition of
highly conserved microbial- or pathogen-associated molecular patterns (MAMPs or PAMPs)
by the plant extracellular pattern recognition receptors (PRRs). This interaction activates
complex host response that results in MAMP-triggered immunity (MTI) or PAMP-triggered
immunity (PTI). The successful pathogen can suppress PTI by delivering effector molecules
into the host. In the presence of resistance (R) protein that matches specifically with the
effector(s), intracellular defense response is activated by the direct or indirect interaction
between them. Effector-mediated activation of R protein results in effector-triggered
immunity (ETI). When a given effector protein is recognized by an R protein, it is referred to
as an avirulence (Avr) protein, as it renders the pathogen non-virulent. The PTI and ETI give
rise to similar responses, although ETI is qualitatively stronger and faster. Such a multi-
layered defense system confers effective resistance against pathogens and the interaction
between plant and pathogen is known as ‘incompatible’. The interaction between Avr and R
protein triggers a rapid and strong response at infection sites. This response is known as the
hypersensitive response (HR) and it involves the induction of programmed cell death (PCD).
The HR also activates different signal transduction pathways which results in the expression
of defense related genes. These induced responses restrict pathogen at the primary infection
sites by the appearance of necrotic local lesions. The pathogen is usually confined to the
lesion and fails to spread into adjacent healthy tissues. Resistance can also be observed where
pathogen replication does not occur, or occurs at essentially undetectable levels in infected
cells. This type of resistance has been termed as ‘extreme resistance’ (ER), ‘cellular
resistance’, or ‘immunity’. The ER response is very rapid and efficient, such that pathogen
accumulation is inhibited before the onset of secondary responses, i.e., HR. The effector
proteins are thought to play a role in virulence on plants in the absence of the cognate R gene.
These interferences results in effector-triggered susceptibility (ETS) and the interaction
between plant and pathogen is termed as ‘compatible’. During such interaction, the pathogen
can infects and replicates in the susceptible host to produce disease.

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Oxidative burst and Reactive Oxygen Species

The ‘oxidative burst’ by the production of reactive oxygen species (ROS) is one of the
earliest cellular responses following pathogen infection. The sequential reduction of
molecular oxygen to superoxide radical (.O2-), hydrogen peroxide (H2O2) and hydroxyl radical
(.OH) are the most predominant ROS produced in plant cell. ROS-scavenging enzymes,
including ascorbate peroxidases (APX), superoxide dismutases (SOD) and catalase (CAT)
maintain ROS homeostasis in different compartments of the plant cell. The incompatible
interaction induces a biphasic ROS accumulation with a low-amplitude in first phase,
followed by a sustained phase of higher magnitude that correlates with disease resistance.
However, virulent pathogens induce only a transient low-amplitude first phase of this
response as a consequence of basal defense. High concentrations of ROS are produced at the
plasma membrane in the vicinity of the pathogen mainly by the activation of NADPH oxidase
and cell wall peroxidases. Although ROS are produced as part of normal metabolism during
photosynthesis and respiration, the high magnitude during incompatible interaction
overwhelm the plant’s own antioxidant system and can directly kills the invading pathogens.
ROS could also contribute to the establishment of physical barriers by cross linking of cell
wall glycoproteins or via oxidative cross linking of lignin and suberin polymers. It can induce
the generation of phytoalexins and secondary metabolites that arrest pathogen growth. Thus,
actively participates in the activation of defense related genes; or indirectly by interaction
with other signaling components like inducing calcium signaling pathway and activating
phosphorylation cascades.

Plant Resistance protein

The genes encoding the specificity determinants of ETI are known as R genes. Most R genes
encode proteins that contain four distinct domains joined by linker regions: a variable amino-
terminal domain, a nucleotide-binding site (NBS) domain, a leucine-rich repeats (LRR)
region and variable carboxy-terminal domains. These NBS-LRR proteins (also called NB-
LRR or NB-ARC-LRR proteins) can be categorized into TIR and non-TIR classes based on
the identity of the amino-terminal domain. The TIR-NBS-LRR (TNL) class of proteins
contains an amino-terminal domain with homology to the Toll receptor in Drosophila and
human interleukin 1 receptor. The non-TIR-NBS-LRR (CNL) class contains α-helical coiled-
coil (CC)–like sequences in their amino-terminal domain. The NBS domain contains several
defined motifs characteristic of the ‘signal transduction ATPases with numerous domains’

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(STAND) family of ATPases. The STAND proteins function as molecular switches in

disease signaling pathways. The LRR domainis participates in protein-protein interactions
and ligand binding. In a resting state, NBS-LRRs proteins are proposed to be bound to ADP.
Intramolecular interactions and association with co-chaperones keep the protein complex in a
competent state. Direct or indirect Avr recognition activates the R protein by a change of
conformation, inducing nucleotide exchange and in some cases a change of cellular
localization. The ATP-bound conformation triggers a second conformational change in the
protein probably by exposing its amino-terminal domain and releasing its signaling potential.
The ATPase activity of the R protein attenuates signaling response and returns the protein in
an inactive ADP-bound conformation.

Systemic acquired resistance

Plants respond to pathogen attack by activating both local and systemic defenses to restrict
pathogen growth and spread. In the uninoculated tissue a state of heightened defense response
is developed that is termed as systemic acquired resistance (SAR). It is similar to acquired
immunity in animals in that it is systemic, long-lasting and provides broad-spectrum
resistance to secondary infection with the same or related pathogen. During SAR a signal is
moved through phloem from the infected tissue to the systemic tissue. Initially, salicylic acid
(SA) was postulated to be this mobile signal because it induces defense responses when
exogenously applied to the plants. However, grafting studies showed that infected, SA-
deficient plant could trigger SAR; such results imply that SA is not the mobile SAR signal.
Later, it has been shown that the SA-derivative methyl salicylate (MeSA) acts as a long-
distance mobile signal for SAR. Upon pathogen infection, endogenous SA level is increased
in the infected tissue; SA methyltransferase 1(SAMT1) convert the SA to MeSA. The MeSA
is transported to the uninoculated systemic tissue and hydrolyzed to SA by the MeSA esterase
activity (MSE) of SA-binding protein 2 (SABP2). The MSE activity of SABP2 is inhibited in
the primary infected tissue by the binding of SA in its active site pocket to facilitate sufficient
level of MeSA accumulation to provide signal for inducing SAR. SAR is characterized by the
increased SA level, changes in redox status, and the dramatic induction of pathogenesis-
related (PR) gene expression in the systemic tissues.

Pathogenesis-related proteins

The PR proteins were first described in the 1970 by van Loon and van Kammen, who
observed accumulation of various novel proteins after infection of tobacco with Tobacco

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Mosaic Virus (TMV). The term PR proteins indicates the proteins that are either absent or
present only at basal concentrations detectable in healthy tissues, but increased upon
pathological conditions and related situations including the application of chemicals that
mimic the effect of pathogen attack (e.g., the plant hormones). There are numerous enzymes
(e.g., phenylalanine ammonia-lyase (PAL), peroxidase, and polyphenoloxidase) that are
present constitutively and increased during most infections, are also referred to as PRs. For
this reason, the term ‘inducible defense-related proteins’ has been introduced for collectively
defined PR proteins. The PR proteins have been classified into 17 structurally and
functionally distinct families. Some of these have enzymatic activity, such as β-1,3-
glucanase, (PR-2), chitinase (PR-3, -4, -8, and -11), endoproteinase (PR-7), peroxidase (PR-
9), or ribonuclease (PR-10) and possess antifungal activity.

2.3.6. Plant hormone signaling

The regulation of the defense network during plant-pathogen interaction depends profoundly
on the action of the hormones. Pathogen infection stimulates the plant to synthesize hormonal
signals depending on the type of invader. Plants produce a wide range of hormones including
auxins, gibberellins (GA), abscisic acid (ABA), cytokinins (CK), salicylic acid (SA), ethylene
(ET), jasmonates (JA), brassinosteroids (BR) and peptide hormones. Among these the
phyotohormones (SA, JA and ET) are known to play major roles in regulating plant defense
responses against wide range of pathogens. SA is generally involved in the activation of
defense responses against biotrophic and hemi-biotrophic pathogens. By contrast, JA and ET
are usually associated with defense against necrotrophic pathogens and herbivorous insects.
These hormones interact in a complex manner by the antagonism of SA and JA, as well as the
synergism between JA and ethylene. Several important regulatory proteins have been found
to associate with contrasting effects on SA and JA signaling and on resistance against
biotrophs and necrotrophs. One of the important regulatory components of SA signaling is
non-expressor of PR genes 1 (NPR1) that are involved in the activation of SA-responsive PR
genes. Several WRKY transcription factors are present in the downstream of NPR1 that play
important role in the regulation of SA-dependent defense responses in plants. These proteins
act as positive regulator of SA-dependent defense and a negative regulator of JA-dependent
defense in order to maintain the balance between these two pathways.

Dr. Subrata Kundu / Plant-pathogen interaction