Presentation points for Plant Immunology

1. How do plant-associated fungi neutralize host ROS?

 Plant innate immune system:
1. Fungal Nox complexes produce ROS for the differentiation of plant-
infecting structures.
2. Host oxidative burst generates superoxide around infection sites, which
needs to be neutralized.
3. Plants have developed innate immune responses to detect pathogens:
a. Pathogen-associated molecular patterns (PAMP)-triggered Immunity
(PTI)
b. Effector-triggered immunity (ETI)
4. Innate immune responses in plants include:
a. Callose deposition
b. Increased expression of pathogenesis-related (PR) genes
c. Oxidative bursts producing ROS
5. Plants recognize extracellular motifs of microbes, leading to rapid release
of ROS during PTI.
6. Plant pathogens have evolved effector proteins to evade basal defense.
7. Plants have resistance (R) proteins that detect effector targets or
perturbations, triggering ETI.
8. ETI results in oxidative bursts and programmed cell death (PCD) known as
the hypersensitive response (HR).
9. Unwanted effects of ETI, such as PCD, hinder biotrophic survival.
10.Pathogens have evolved effector proteins to bypass detection from R
proteins.
11.Co-evolution between plants and pathogens forms the basis of the zig-zag
model of plant pathology.
12.Oxidative bursts are not effective against necrotrophic fungi B. cinerea and
Sclerotinia sclerotiorum.
13.Hemibiotrophs like Septoria tritici and M. oryzae can tolerate oxidative
bursts.
14.Fungi have mechanisms to deal with host ROS and maintain redox balance
during host infection.
 Transcriptional control of ROS catabolism
1. Fungi possess antioxidation systems to maintain redox homeostasis and
counteract host ROS.
2. The thioredoxin system includes thioredoxins (Trx) and thioredoxin
reductase (TrxR).
3. Thioredoxins are redox proteins with reversible activity, cycling between
oxidized disulfide (Trx-S2) and reduced dithiol [Trx-(SH)2].
4. Glutaredoxin proteins, part of the glutathione system, also contribute to
ROS scavenging.
5. Glutaredoxins are non-enzymatically reduced by the antioxidant
glutathione (GSH), which is recycled by glutathione reductase.
6. Other enzymes involved in ROS detoxification include superoxide
dismutase (SOD), peroxidases, and catalases.
7. SOD catalyzes the dismutation of the superoxide radical (O2%) to H2O2.
8. Catalases and glutaredoxins further convert H2O2 to H2O or neutralize it.
9. CuZnSOD is the primary O2%− cellular detoxifier in the cytosol, nucleus,
and mitochondrial inner membrane.
10.Catalase B (CatB) has varying effects on pathogenicity in many
phytopathogenic fungi.
11.Yap1 is a well-studied transcriptional regulator of antioxidation genes in
Saccharomyces cerevisiae.
12.Yap1 is a basic leucine zipper (bZIP) transcription factor that activates
enzymatic and non-enzymatic antioxidation genes.
13.Yap1 preferentially binds to specific sequences in the promoter regions of
glutathione and thioredoxin system genes.
14.Under normal conditions, Crm1, a nuclear export protein, prevents Yap1
nuclear accumulation.
15.During oxidative stress, Yap1 cysteine residues are oxidized, leading to its
dissociation from Crm1 and accumulation in the nucleus.
16.Yap1 in the nucleus binds to the promoters of oxidative stress response
genes encoding antioxidants and thiol-reducing pathways.
17.Yeast AP1-like transcription factors, including Yap1, are important for
fungal phytopathogens' response to oxidative stress.

 Neutralizing the host oxidative burst

1. The role of Yap1 homologs and oxidative stress response components in
plant infection has been studied in fungi with different lifestyles.
2. Biotrophic fungi are symptom-causing fungi that rely on living hosts for
their nutrition.
3. Endophytes reside in intimate contact with living host cells without causing
visible symptoms.
4. Both biotrophic fungi and endophytes must handle host-generated
reactive oxygen species (ROS).
5. Necrotrophic fungi feed on host cells after killing them and neutralize host
oxidative bursts to avoid programmed cell death (PCD) in plants.
6. Necrotrophic fungi may not benefit from triggering host PCD as an
infection strategy.
7. Hemibiotrophs, such as the rice and wheat blast fungus M. oryzae, initially
colonize living host cells as asymptomatic biotrophs before switching to
necrotrophy.
8. The early biotrophic growth phases of hemibiotrophs require host defense
suppression and ROS neutralization.
9. Despite the common occurrence of host oxidative bursts in fungal-plant
interactions, research has primarily focused on the host response.
10.A comprehensive understanding of antioxidation strategies employed by
fungi is still lacking.
11.There have been some advancements in studying the antioxidation
strategies of fungi.

 Biotrophs and endophyte

1. Endophytes and other beneficial symbionts, such as mycorrhizal fungi,
experience an oxidative burst upon colonizing their host plants.
2. The oxidative burst is believed to be beneficial for successful colonization
in symbiotic relationships.
3. Upregulation of antioxidation genes has been observed in arbuscular
mycorrhizal fungi, leading to increased stress tolerance in plant hosts.
4. The roles of antioxidation enzymes and genes in fungal partners have
received less attention compared to plants in symbiotic relationships.
5. In the ericoid mycorrhizal fungus Oidiodendron maius, loss of SOD1
activity increased ROS sensitivity and reduced the fungus's capacity to
colonize host roots, highlighting the importance of redox homeostasis in
mycorrhization.
6. Understanding the genetic arsenal required for successful plant
colonization through molecular analysis is crucial.
7. The characterization of genes and enzymes involved in overcoming host
oxidative bursts by biotrophic pathogens is limited.
8. The Yap1 homolog UmYap1 in the biotrophic maize pathogen Ustilago
maydis is required for virulence and nuclear localization upon H2O2
exposure, suggesting the importance of ROS neutralization for
pathogenicity.
9. In the symbiotic fungus Epichloë festucae, the Yap1 homolog YapA is
involved in H2O2 sensitivity in conidia but is not required for plant
infection.
10.Catalase, a ROS-scavenging enzyme, has been studied in biotrophic fungi.
In C. purpurea, the catalase component CATB is not essential for virulence
due to functional overlap with other catalases, but the absence of all
catalase activity reduces virulence.
11.Further investigation of the molecular mechanisms involved in combating
host oxidative bursts in biotrophic, and endophytic fungi is necessary to
understand antioxidation strategies in beneficial relationships.

 Hemi-Biotrophs
1. The hemibiotrophic fungus M. oryzae possesses a range of antioxidation
genes to counteract oxidative stress from its rice host.
2. The transcriptional regulator MoAp1 is necessary for biotrophic growth and
conidiogenesis in M. oryzae.
3. The thioredoxin and glutathione antioxidation systems play crucial roles in
M. oryzae pathogenicity.
4. Specific components of the thioredoxin system (Tpx1, Trr1, and Trx2) are
required for intracellular ROS metabolism and in planta proliferation of M.
oryzae.
5. Glutathione reductase (Gtr1) is necessary for both in planta proliferation
and neutralizing host-derived ROS accumulation in M. oryzae.
6. The Tps1 enzyme, involved in glucose-6-phosphate/NADPH sensing,
regulates the expression of NADPH-dependent enzymes, including those
of the thioredoxin and glutathione antioxidation system in M. oryzae.
7. The NMO2 gene in M. oryzae, regulated by Tps1, is required for protection
against reactive nitrogen species (RNS) and mediates nitrooxidative stress.
8. Loss of NMO2 triggers a strong host oxidative burst during rice cell
colonization, leading to rice innate immune responses and impaired fungal
growth.
9. The relationship between the Yap1 homolog MoAp1 and Tps1 sugar
signaling in M. oryzae is unknown but MoAp1 is required for virulence.
10.Catalase enzymes, CatB and CpxB, play distinct roles in M. oryzae. CatB
strengthens the cell wall and appressoria melanization, while CpxB
neutralizes host-derived ROS.
11.M. oryzae has robust antioxidation capabilities, detoxifying higher
concentrations of ROS than those resulting from host oxidative bursts.
12.The loss of MoSir2 function in M. oryzae leads to elevated defense
responses due to the inability to neutralize host ROS, resulting in the
trapping of the mutant without killing it.
13.Dealing with host ROS, even in compatible interactions, is a critical step for
M. oryzae to avoid triggering host defenses.

 Necrotrophs:
1. Necrotrophic fungi like B. cinerea and S. sclerotiorum may utilize host
reactive oxygen species (ROS) accumulation to trigger hypersensitive
response (HR) and kill host cells during infection.
2. The aggressiveness of B. cinerea isolates corresponds to the intensity of the
oxidative burst it produces.
3. The Yap1 homolog (Bap1) in B. cinerea is involved in ROS detoxification in
axenic culture but is not required for in planta proliferation.
4. The Yap1 homolog (ChAP1) in Cochliobolus heterostrophus is dispensable
for virulence but required for oxidative stress tolerance.
5. The AaAP1 gene in Alternaria alternata is required for virulence, unlike in
other necrotrophs.
6. Superoxide dismutase (SOD) is required for host infection by B. cinerea,
potentially due to its role in either ROS neutralization or H2O2 production.
7. The thioredoxin system, rather than the glutathione system, is essential for
B. cinerea survival in its host, with Trx1 and Trr1 being required for
infection.
8. Glutathione reductase, responsible for recycling oxidized glutathione, has a
minor role in B. cinerea development and pathogenicity compared to
thioredoxins.
9. The role of specific antioxidation genes and enzymes in pathogenicity can
vary within fungal-host interactions.
10.The function of antioxidation components may differ between detoxifying
extracellular host-derived ROS and intracellular ROS.
11.S. sclerotiorum requires SOD for pathogenicity due to its role in oxalate
production, while catalases are required for pathogenicity but not for
scavenging extracellular host-derived ROS.
12.The oxidative stress encountered in the host may induce Nox-catalyzed
differentiation processes in B. cinerea, which are necessary for penetration
or colonization.

These points highlight the contrasting strategies employed by necrotrophic

fungi during host interactions regarding the role of host ROS. While some
necrotrophs utilize and potentially contribute to host ROS accumulation to
trigger host cell death, others employ antioxidation systems to neutralize ROS
or have other mechanisms for dealing with oxidative stress during infection.
The specific roles of antioxidation genes and enzymes can vary, and further
research is needed to understand their precise functions and the interplay
between fungal and host oxidative stress responses.