PLANT DEFENSES AGAINST BIOTIC STRESSES

Plant Physiology, Fifth Edition (2010), L. Taiz & E. Zeiger

Chapter 13 Secondary Metabolites and Plant Defense

Biochemistry & Molecular Biology of Plants (2000), B.B. Buchanan, W. Gruissen, R. J.
Jones. Chapter 21 Responses to Plant Pathogens
FISIOLOGIA VEGETAL COMPLEMENTAR
(2013/2014)

Plants must continuously defend themselves against
attack from other organisms, including vertebrates
herbivores and invertebrates like phytophagous insects,
as well as plant pathogens, including fungi, bacteria,
viruses, and so on.

Because their immobility precludes escape, each plant
organism possess both preformed (constitutive) and an
inducible defense capacity.

In wild plant populations, most plant are healthy most of the time. If disease
does occur it is usually restricted to only a few plants and affects only a small
amount of tissue. Disease, the outcome of a successful infection, rarely kills a
plant. Natural selection probably acts to curtail fatal pathogen toxicity to plants
diseases that keep a host alive longer may permit more reproduction of
pathogen.
PLANT DEFENSES AGAINST INSECT HERBIVORES
Plants have developed a wide variety of defensive strategies against
insect herbivory which can be divided into two categories: (a)
constitutive defenses; (b) induced defenses.

a) Constitutive defenses defensive mechanisms always present in the
plant. They are often species-specific and may exist as stored
compounds, conjugated compounds (to reduce toxicity), or precursors of
active compounds that can easily be activated if the plant is damaged.

a) Induced defenses are initiated only after actual damage occurs. They
include the production of defensive proteins such as lectins and protease
inhibitors as well as the production of toxic secondary metabolites.

In some cases the same insecticidal defensive compounds are
involved in both constitutive and induced defensive responses.
Cyanogenic glycosides release the poison hydrogen cyanide
Glucosinolates release volatile toxins.

Cyanogenic glycosides and Glucosinolates are nitrogen non-alkaloid protective compounds
which themselves are not toxic but are readily broken down to give off poisons, some of
which are volatile, when the plant is crushed.

Cyanogenic glycosides release the well-known poisonous gas hydrogen cyanide (HCN) in a
two-step enzymatic process:
1 In the first step the sugar is cleaved by a glycosidase (Figure 13.17)
2 - In the second step the resulting hydrolysis product, called -hydroxynitrile or
cyanohydrine, is decomposed with release of HCN .
Figure 13.17. Enzyme-catalysed hydrolysis of cyanogenic glycosides to release hydrogen
cyanide. R and R represents various alkyl or aryl substituents.

In Amygdalin (the common cyanogenic glycoside found in seeds of almonds, apricots, cherries
and peaches) R is phenyl, R is hydrogen, and the sugar is the disaccharide -gentiobiose.
As cyanogenic glycosides and the respective degradative enzymes are
separated in different subcelluar compartments or in different tissues
they are not normaly broken down in the intact plant.

However, when the leaf is damaged, as during herbivore feeding , the
cell contents of different tissues mix, and HCN forms.

Cyanogenic glycosides are widely distributed in the plant kingdom and
are found in many legumes, grasses and species of the rose family.

HCN is a fast-acting toxin that inhibits metalloproteins such as the
iron-containing cytochrome oxydase, a key enzyme of mitochondrial
respiration.

The presence of cyanogenic glycosides deters feeding by insects and
other herbivores.

Some herbivores, however, have adapted to feed on cyanogenic plants and can tolerate large
doses of HCN.
A second class of plant glycosides the glucosinolates, or mustard oil glycosides,
break down to release defensive substances.

Found principally in the Brassicaceae and related plant families, glucosinolates
break down to produce the compounds responsible for the smell and taste of
vegetables such as cabbage, brocoli, and radishes.

Glucosinolate breakdown is catalyzed by a hydrolytic enzyme thioglucosidase or
myrosinase that cleaves glucose from its bond with the sulfur atom (Fig. 13.18).

The resulting aglycone loses the sulfate and rearranges itself to give pungent and
chemically reactive products, including isothiocyanates and nitriles, depending of
the conditions of hydrolysis, which functions as toxins and hervibory repellents.
Figure 13.18 Hydrolysis of glucosinolates into mustard-smelling volatiles.
R represents various alkyl or aryl substituents. In sinigrin, (a major glucosinolate of black
mustard seeds) R is CH=CH-CH.
Non-protein amino acids toxic to hervibores
Many plants produce non-protein aminoacids that are present in free form (they
are not incorporated in plant proteins) and act as defensive substances.

Due to similarity with some protein amino acids, such unusual amino acids block
the synthesis or uptake of protein amino acids or can be mistakenly incorporated
into proteins, as it happens with canavanin and azetidine-2-carboxylic acid which
have structures similar to those of arginine and proline respectively (Figure 13.19).

Proteins incorporating such unusual amino acids are non-functional what explain
their toxicity to ruminants.
Figure 13.19. Nonprotein
amino acids and their protein
amino acid analogues.

The nonprotein amino acids are not
incorporated into proteins, but are
defensive compounds found in free
form in plant cells. Their activity
ranges from interference with the
uptake of amino acids to the
disruption of translation.
INDUCED PLANT DEFENSES AGAINST INSECT HERBIVORES
In principle, induced defenses requires a smaller investment of plant
resouces than constitutive defenses, but they must be activated
quickly to be effective.

Plants can recognize specific components of insect saliva

The plant response to damage by insect herbivores involves both a wound response and the
recognition of certain insect-derived compounds referred to as ELICITORS certain molecules
in insect saliva can serve as enhancers of the stimulus of the wound responses.

In addition, such insect-derived elicitors can trigger signaling pathways systematically, thereby
initiating defensive responses in distant regions of the plant in anticipation of further damage.

The first elicitors identified in insect saliva were Fatty acid-aminoacid conjugates (or fatty
acid amides). The biosynthesis of these conjugates depends on the plant as the source of the
fatty acids linolenic (18:3) and linoleic (18:2) fatty acids which are subsequently coupled to an
amino acid in the insect gut.

Plants, then recognize these elicitors and activate complex signal transduction pathway that
induces their defenses.
Jasmonic acid activates many plant
defences

A major signaling pathway involved in most
plant defenses against insect herbivores is
the octadecenoic pathway which leads to
production of a plant hormone called
JASMONIC ACID (JA or jasmonate) Fig. 13.20.





Figure 13.20. Steps in the pathway for
conversion of linolenic (18:3) into jasmonic
acid. The first three enzymatic steps occur
in the chloroplast, resulting in the cyclized
product 12-oxo-phytodienoic acid. This
intermediate is transported to peroxisome,
where it is first reduced and then
converted into jasmonic acid by -oxidation
Jasmonic acid (JA) is known to induce the transcription of
host genes involved in defensive metabolism.

Among the genes it induces are those that encode key enzymes in
all the major pathways for secondary metabolite biosynthesis.

It has been shown that JA acts through a conserved signaling
mechanism that bears close resemblance to those described for
other plant hormones such as auxin and gibberelins (Figure 13.21).

However, in contrast to those hormonal signals, JA first needs to be
activated by conversion into an amino acid conjugate by JAR
proteins, which belong to a family of carboxylic acid-conjugating
enzymes.

The enzyme JAR1, for example, exhibits a hight substract specificity
for JA and Isoleucine (JA-Ile) and appears to be of special
importance for JA-dependent defensive signaling.
Figure 13.21.

A model for JA signaling. In contrast to other hormonal
signals, JA needs to be conjugated first to an amino acid
(here isoleucine) by the enzyme JAR1. The resulting
jasmonic acid-isoleucine (JA-Ile) conjugate then binds to
COI1 as part of a SCFCOI1 protein complex. This complex
targets JAZ, a repressor of transcription, leading to
polyubiquitination and subsequent degradation of this
protein in the 26S proteasome. Transcription factors such
as MYC2 then initiate transcription of jasmonic acid-
dependent genes encoding defensive proteins. In addition
JAZ genes are activated, thereby providing a negative
feedback mechanism for this signaling pathway.
Some plant proteins inhibit herbivore digestion.
Among the diverse components of plant defense mechanisms induced
by jasmonic acid are proteins that interfere with herbivore digestion
namely:

(a) -amylase inhibitors that block the action of starch-digesting enzyme
-amylase (in some legumes)
(b)Lectins defensive proteins that bind to carbohydrates or
carbohydrate-containing proteins. After ingestion by an herbivore,
lectins bind to the epithelial cells lining the digestive tract and
interfere with nutrient absorption
(c) Protease inhibitors block the action of herbivore proteolytic
enzymes (proteases). After entering the herbivores digeste tract ,
they hinder protein digestion by binding tightly and specifically to the
active site of proteolytic enzymes such trypsin and chymotrypsin
(found in legumes, tomatoes, and other plants).
Damage by insect herbivores induces systemic defenses
In tomatoes. Insect feeding leads to the rapid accumulation of protease inhibitors throughout the
plant even in undamaged areas far from the initial feeding site. The systematic production of
protease inhibitors in young tomato plants is triggered by a complex sequence of events (Fig. 13.22).
Figure 13.22.
Proposed system in signaling pathway for the rapid induction
of protease inhibitor biosynthesis in wounded tomato plants.

(1) Wounded tomato leaves (boton of the figure)
synthesize prosystemin (200 amino acids) in phloem
parenchyma cells, and the prosystemin is
proteolytically processed to systemin (18 amino acids)
(2) Systemin is released from phloem parenchyma
cells and binds to receptors on the plasma
membrane of adjacent companion cells.
(3) This binding activates a signaling cascade involving
phospholipase A (PLA) and mitogen-activated
protein (MAP) kinases which results in the
biosynthesis of JA.
(4) JA is then transported via sieve elements, possibli
in a conjugated form (JA-X), to wound leaves.
(5) There, JA initiates a signaling pathway in target
mesophyll cells, resulting in the expression of genes
that encode protease inhibitors.
(6) Plasmodesmata facilitate the spread of the signal.

Herbivore-induced volatiles have complex
ecological functions
The induction and release of volatile organic compounds, also
called volatiles, in response to insect herbivore damage provides
an excellent example of the complex ecological functions os
secondary metabolites in Nature.

The mixture of volatile molecules includes representatives from
the three major classes of secondary metabolites: terpenes,
phenolics, and alkaloids.

Additionally in response to mechanical damage, all plants emit
lipid-derived products such as green-leaf volatiles a mixture of
six-carbon aldehydes, alcohols, and esters.
Plant Defenses against Pathogens
Plant Defenses against Pathogens
Even though plants lack an immune system as complex as that in animals, plants
are surprisingly resistent to diseases caused by fungi, bacteria, viruses, and
nematodes that are ever present in the environment.
Pathogens have developed various strategies to invad host plants

(a)Some pathogens penetrate the cuticule and cell wall directly by
secreting lytic enzymes, which digest mecanical barriers;
(b)Others enter the plant through natural openings like stomata
and lenticels;
(c) A thirth category invades the plant through wounding sites, for
example those caused by insect herbivores. Additionaly, many
virus, as well as other types of pathogens are transfered by
insect herbivores, which serve as vectors, and invade the plant
from the insect feeding site.

Once inside the plant, pathogens generally use one of the three main
attack strategies to utilize the host plant as a substrate for their own
progression:

(a)Necrotrophic pathogens attack their host plant by secreting cell
wall-degrading enzymes or toxins, which eventually kill the affected
plant cells leading to massive tissue laceration. This dead tissue is
then colonized by pathogen and serves as food source.
(b)Biotrophic pathogens. After infection, the plant tissue remains
largely alive, and only minimal cell damage can be observed while
the pathogen continues to feed on substrates provided by host.
(c)Hemibiotrophic pathogens. After an biotrophic initial stage, the
pathogen acts as nectrotrophic pathogen.

Although these invasion and infection strategies are individually
successful, plant disease epidemics are rare in natural ecosystems.
This is because plants have developed defense strategies against this
diverse array of pathogens.
Some antimicrobial compounds are synthesized before
pathogen attack

Several classes of secondary metabolites have strong antimicrobial
activity acting as defenses against pathogens in the intact plant.
Among these, are saponins, a group of triterpenes thought to
disrupt fungal membranes by binding to sterols
Infection induces additional antipathogen defenses

After being infected by a pathogen, plants deploy a broad spectrum
of defenses against invading microbes.
A common defense is the hypersensitive response, in which cells
immediately surrounding the infection site die rapidly, depriving the
pathogen of nutrients and preventing its spread. After a successful
hypersensitive response, a small region of dead tissue is left at the
site of the attempeted invasion, but the rest of plant is unaffected.
The hypersensitive response is often preceded by the rapid accumulation of ROS and NO. Cells
in the vicinity of the infection synthesize a burst of toxic compounds formed by the reduction
of molecular oxygen, including the superoxide anion (O), hydrogen peroxide (HO), and the
hydroxyl radical (HO). A NADPH-dependent oxidase located at the plasma membrane is
thought to produce O, which in turn is converted into HO and HO (Figure 13-23).
Figure 13.23. Many types of antipathogen defences are induced by infection. Fragments of
pathogen molecules called elicitors initiate a complex signaling pathway leading to the
activation of defense responses. A burst of oxidation activity and nitric oxide production
stimulates the hypersensitive response and other defense mechanism.
Ca is necessary for the activation of some defenses, while it is also a negative regulator of
salicylic acid biosynthesis.
Arginine
MAMP receptor
or
R gene product
The hydroxyl radical (HO) is the strongest oxidant of these reactive
oxygen species (ROS) and can initiate radical chain reactions with a
range of organic molecules, leading to lipid peroxidation, enzyme
inactivation, and nucleic acid degradation.

Reactive oxygen species may contribute to host cell death as part of the
hypersensitive response or act to kill the pathogen directly.

A rapid spike of NO production from the amino acid arginine by the NO
synthase, accompanies the oxidation burst in infected leaves in a way
dependent of an increase in cytosolic Ca.
NO acts as a second messenger in many signaling pathways in animals and
plants.
NO and ROS is required for the activation of hypersensitive response.

Many plants react to fungal or bacterial invasion by synthesizing (a) lignin or
callose that serve as barriers to the pathogen; (b) phytoalexins; and/or (c)
hydrolytic enzymes that attack the cell wall of the pathogen.

Hydrolases as, glucanases, chitinases and other Pathogenesis Related (PR)
proteins are also produced.
Phytoalexins often increase after pathogen attack

Phytoalexin production appears to be a common mechanism of resistence
to pathogen microbes in a wide range of plants. However, different plant
families employ different types of secondary metabolites as phytoalexins.
For example, in leguminous plants, such as alfalfa and soybean,
isoflavonoids are common phytoalexins, whereas in solanaceous plants,
such as potato, tobacco, and tomato, various sesquiterpenes are produced
as phytoalexins (Figure 13.24).
Figure 13.24

Structure of some phytoalexins
found in two different plant
families
Some plants recognize specific pathogen-derived substances

Resistant plants respond more rapidly and more vigorously to pathogens than
susceptible plants.

A fist line of resistance is provided by a system that recognizes broad categories of
pathogens. Plants have a variety of receptors that recognize so-called microbe-
associated general molecular patterns (MAMPs). These elicitors are evolutionary
conserved pathogen-derived molecules such as structural elements from the fungal
cell wall or the bacterial flagellum.

MAMPs are recognized by specific receptors which then activates specific plant
defensive responses, including massive phytoalexin production (Figure13.23).
With one only receptor, a plant can recognize a complete taxonomic group that
features a particular MAMP. This form of defensive strategy is known as innat
immunity.

A second system that provides specific resistance to pathogens is mediated through
the interaction of plant R gene (or Resistance gene) products and pathogen-derived
avirulent (Avr) gene products. Most R genes are through to encode protein receptors
that recognize specific molecules originating from pathogens. Binding of these
elicitors to the receptors alerts the plant to the pathogens presence (Fig. 13.23).
Exposure to elicitors induces a signal transduction cascade

Within a few minutes after pathogenic elicitors have been
recognized by an R gene product or MAMP receptor, complex
signaling pathways are set in motion that eventually lead to defensive
responses (Figure 13.23).

A common early element of these cascades is a transient change in
the ion permeability of the plasma membrane.

R-gene receptor activation stimulates an influx of Ca and H ions
into the cell and an efflux of K and Cl ions.

The influx of Ca activates the oxidative burst that may act directly in
defense, as well as signaling other defensive reactions.

Other components of pathogen-stimulated signal transduction
pathways include NO, MAP kinases, Ca-dependent protein kinases,
and several hormones, as JA, ethylene and salicylic acid (SA).

A single encounter with a pathogen may increase
resistance to future attacks.

When a plant survives infection by a pathogen at one site, it often
develops increased resistance to subsequent attacks at sites throught
the plant and enjoys protection against a wide range of pathogenic
species.

This phenomenon, called systemic acquired resistance (SAR),
develops over several days following initial infection (Figure 13.25).

SAR appears to result from increased levels of certain PR proteins
including chitinases and other hydrolytic enzymes.

Although the mechanism of SAR induction is still unknown, one of the
endogenous signals involved is likely to be salicylic acid whose accumulation
increases dramatically in the zone of infection after the initial attack.
Figure 13.25
Initial pathogen infection may
increase resistance to future
pathogen attack through
development os systemic
acquired resistence (SAR).
From the infection site, SAR is transmitted through the phloem to other parts of
the plant, resulting in increased resistance through the plant.

Salicylic acid and its methyl ester increase significantly in this process and cause
the production of pathogenesis-related (PR) proteins (for example, chitinases and
other hydrolytic enzymes).

Methyl salicylate is often released during SAR and may serve as a SAR-inducing
volatile signal in neigboring plants.
Interactions of plants with non-pathogenic bacteria
can trigger induced systemic resistance (ISR).




In contrast to SAR, which occurs as a consequence of actual pathogen
infection, Induced systemic resistance (ISR) is activated by non-
pathogenic microbes (Figure 13.26).
Figure 13.26
Exposure to nonpathogenic
microorganisms may increase
resistance to future pathogen
attack through development of
induced systemic resistance
(ISR).
Non-pathogenic microorganisms such as rhizobacteria activate signaling pathways
involving jasmonic acid and ethylene that trigger ISR throughout the plant.

Rather than inactivating immediate defensive measures, ISR is characterized by an
increased level of preparedness against pathogen attack.
Ways in which plant pathogens
cause disease.
Roots and shoots of all plants come into intimate
contact with plant pathogens.

Each pathogen has evolved a specific way to
invade plants: (a) penetrating surface layers by
enzymatic attack; (b) through natural openings; (c)
through previously wounded tissue. Once inside
the plant the the attack strategy can be by: (a)
necrotrophy; (b) biotrophy; (c) hemibiotrophy
(figure 21.3; Table 21.1).
Pathogenesis: process of infection,
colonization, and pathogen reproduction.
Virulent: pathogen strain that causes disease
Figure 21.3 Most microbes attack only a specific
part of the plant and produce characteristic
disease symptoms, such as mosaic, necrosis,
spotting, wilting, or enlarged roots. Tomato plants
are attacked by more than 100 different
pathogenic microorganisms
Fungal plant pathogens use a wide range of pathogenesis
strategies
Necrotrophic species that produce cell wall-degradding species tend
to attack a broad range of plant species as it is the cases of Pythium
sp. and Botrytis sp. That attack more than 1000 plants.
Figure 21.4

Botrytis cinerea, the gray mold fungus,
sporulating on grapes. This necrotroph
secretes large numbers of cell wall-
degrading enzymes and thereby
destroys plant tissue inadvance of the
colonizing hyphae
Some necrotrophs produce host-selective toxins that are active in only a few plant
species. Each toxin has a highly specific mode of action, inactivating just a single
plant enzyme. For example , the HC-toxin produced by the maize fungal pathogen
Cochilobolus carbonum inhibits the activity of histone deacetylase, an enzyme that
is required for the activation of plant defense genes (Figure 21.5).
Figure 21.5
(1) The maize pathogen Cochilobolus carbonum
secretes HC-toxin and numerous nonspecific fungal
elicitors that trigger plant defense responses. (2) HC-
toxin is hypothesized to enter the responding plant
cells, where it inhibits histone deacetylase enzyme
activity. Histone deacetylase is responsible for the
reversible deacetylation of core histones (H3 and H4)
assembled in the chromatin. Inhibition of histone
deacetylase activity is believed to interfere with
transcription of maize defense genes and thus
favours fungal growth and disease development.

Hm1-resistant maize plants produce an HC-toxin reductase
(HCTR), which detoxifies HC-toxin by reducing the carbonyl
group on the side chain of L-Aeo(2-amino-9,10-epoxy-o-
oxo-decanoic acid), an unusual epoxidated fatty amino acid.
Biotrophic fungi usually exhibit a high degree of specialization for individual
plant species. To utilize living cells as a food substrate, these fungi often
penetrate the rigid cell wall by forming first a penetration peg and then a
haustorium, which causes invagination of plasma membrane. This
specialized feeding structure increases surface contact between the two
organisms, thus maximizing nutrient and water flow to favour fungal growth
(Figure 21.6).
(A)
(B)
Figure 21.6
(A) Diagram of a fungal houstorium, a
structure that facilitates the transfer of
nutrients from a living plant cell to the
pathogen. The extracelular houstorial
matrix contains products of both fungal
and plant origin.
(B) Scanning electron micrograph of a
houstorium of the biotrophic barley
mildew fungus Erysiphe graminis f.sp.
Hordei (Blumeria graminis) inside the
epidermal cell of barley.
A few biotrophic fungi as the tomato leaf mold pathogen
Cladosporium fulvum do not form haustoria but instead grow
exclusively outside the plant cell wall within the intercellular air space
(apoplast).
Hemibiotrophic fungi sequentially deploy a biotrophic and then a
necrotrophic mode of nutrition. The switch is usually triggered by
increasing nutritional demands as the fungal biomass increases.
Some of the most devasting phytopathogenic species fall into this
category. For example, the organism Phytophthora infestans, which
causes late blight disease of potato (Figure 21.7).
Bacterial pathogens of plants and animals appear to use
similar molecular mechanisms for invading tissues.

Phytopatogenic bacteria specialize in colonizing the appoplast to cause rots, spots,
vascular wilts, cankers and blights. Most are Gram-negative. Two features
characterize bacteria-plant relationships. First, during their parasitic life, most
bacteria reside within the intercellular spaces of the various plant organs or in the
xylem. Second, many cause considerable plant tissue damage by secreting either
toxins, extracellular polysacharides (EPSs), or cell wall-degrading enzymes at
some stage during pathogenesis (Figure 21.8).
Several bacterial genes, referred to collectively as the hypersensitive response and
pathogenicity cluster, are absolutely required for bacterial pathogenesis. Many hrp
gene sequences from bacteria are very similar to genes required for pathogenesis in
bacteria that infect animals, which sugests that these distinct pathogens utilize
similar virulence strategies. One known strain of Pseudomonas aeruginosa is
capable of causing disease in both Arabidopsis and mice (Figure 21.9)
Box 21.1 Agrobacterium-mediated T-DNA transfer, a mechanism used
in pathogenesis, has become a tool for transforming plant genomes.
Some plant pathogenic nematodes modify the metabolism of root
cells, inducing the plant to form specialized feeding structures.
More than 20 genera of nematodes cause plant diseases by infecting nearly
always the plant root system. These root infections cause often substantial
modification in root architecture and can dramatically alter the entire plant
metabolism.

All parasitic nematode species are obligate biotrophs, and all possess a
hollow feeding stylet capable of penetrating plant cell walls (Figure 21.13A).

Ectoparasitic nematodes feed exclusivley at root surface, whereas
endoparasitic species invade root tissues and spend a large portion of their
cycle in intimate association with root cells.

The most damaging nematodes are: (a) the cyst nematodes (genera
Heterodera and Globodera) and: (b) the root-knot nematodes (Genus
Meloidogyne), both from the same family Heteroderidae.
Nematodes life cycles

(1) The life cycle of the endoparasitic nematodes commences when dormant eggs
perceive an as yet uncharacterized chemical signal released by plant roots.
(2) Receipt of the signal induces the eggs to hatch, releasing juvenile nematodes.
(3) The motile second-stage juveniles then penetrate the roots and migrate to
vascular tissue.
(4) Once feeding is initiated, the nematodes lose motility and becomes sedentary.
(5) To commence feeding, the juvenile cyst nematodes push their stylets through
the plant cell wall, but not the plasma membrane, and release glandular secretions
into selected cells.
(6) Molecules in the secreted fluid induce rapid modifications to the cytoplasm of
the plant cell, and the metabolic activity of the plant cell incrases markedly.
(7) In addition nematodes triggers the dissolution of plant cell walls provocating
firstly the increase of symplastic conections of adjacent cells and later their
protoplast fusion resulting in the formation of feeding structure made from
recruitment of as many as 200 semifused cells.

In contrast, the feeding root-knot nematode induces both mitoses uncoupled from
cytokinesis and DNA endo-reduplication producing abnormal cortical cell growth
resulting in giant cells (Figure 21.13B).
Figure 21.13 Plant root-endoparasitic nematode interaction. (A) Diagram of the anterior of a
nematode, which uses a stylet to enter a plant cell and to pump secretions from the amphid
glands into the selected plant cell. Subsequently, inside the invaded plant cell a new feeding
tube is synthesized, which the nematode uses to extract nutrients specifically from the
modified cell cytoplasm. In each plant cell selected, hundreds of discarded feeding tubes can
accumulate (B) Transverse section of tomato root, showing a root-knot nematode and several
giant cells that have formed at the feeding site of this now sedentary feeding nematode.
Feeding arthropods not only damage plants directly but also
facilitate colonization by viral, bacteria, and fungal pathogens.

Two broad categories of insects are recognized: those that chew
and those that suck sap.

Chewing insects cause the more spectacular plant tissue damage
(Figures 21.14).
The sap-sucking insects use a specialized
mouth part, the stylet, to locate,
penetrate, and drain sap from the phoem
sieve elements of the plants vascular
sieve.

Sap-sucking insects are extremely efective
virus, bacteria, and fungal pathogen
vectors (Figure 21.15).
Genetic basis of plant-pathogen interactions
Besides the preformed defences that involve numerous secondary
metabolites and besides a first line of resistance provided by a system
that recognizes broad categories of pathogens, based in a variety of
receptors that recognize the so-called microbe-associated general
molecular patterns (MAMPs), plants show a second defence system
that provides specific resistance to pathogens which is mediated
through the interaction of plant R gene (R) (or Resistance gene)
products and pathogen-derived avirulent (Avr) gene products.

Such second system of disease resistance is usually mediated by
dominant genes, although some recessive resistance genes also exist.

To explain such mechanism, Harold H. Flor (1940s) proposed
the GENE-FOR-GENE model (Figure21.18).
GENE-FOR-GENE model (For the most biotrophic plant-pathogen interactions)
This model predicts that plant
resistance will occur only when
a plant possesses a dominant
resistance gene (R) and the
pathogen expresses a
complementary dominant
avirulence gene (Avr)
For pathogens that deploy host-selective toxins for successful
pathogenesis, a different model is proposed. Pathogen virulence must
be dominant because a functional toxin or enzyme (or both) must be
produced to cause disease. (Figure 21.19).
Plant resistance, in turn, is predominantly
inherited as a dominant trait and is
achieved through enzymatic detoxification
or through loss or alteration of the toxin in
the pathogen.
The Avr gene products have no signal peptide sequence and so cannot be exported outside
the bacterial cell by the SecA secretory pathway.

The mechanism utilized termed Type III secretory pathway involves a protein complex that
spans both inner- and outerbacterial membranes. Specific protein subcomponents of the
complex are responsible for translocating each protein to be exported, first across the inner
bacterial membrane and then across the outer one and then through a projecting pilus to
complete its transfer into the host plant cell.

How R and Avr gene products activate plant defense responses is not yet understood

Each R gene product is thought to possess two functions: recognition of the corresponding
Avr-derived signal, and activation of downstream signaling pathways to trigger the complex
defense response.

Hypersensitive Response and
Pathogenicity proteins (Hrp) of plant
pathogenic bacteria are also involved.
Bacterial Hrp genes are required for
basic pathogenicity, i.e., disease
formation in a susceptible plant host,
as well as for the Avr gene product to
cause a hypersensitive response in a
resistant plant host.
END
QUESTIONRIO
1 - O que so defesas constitutivas das plantas?!
2 - Como explica que as defesas constitutivas das plantas no provoquem danos nas
prprias plantas que os produzem?!
3 O que so defesas Induzidas (ou induzveis) das plantas?!
4 D 4 exemplos de meios de defesa de tipo constitutivo
5 D 4 exemplos de meios de defesa de tipo induzvel
6 Explique sucintamente a aco de defesa contra insectos baseada nos glicsidos
cianognicos
7 De que consiste a toxicidade do cianeto de hidrognio?
8 - Explique sucintamente a aco de defesa contra insectos baseada nos glucosinolatos
9 Explique sucintamente a aco de defesa contra insectos baseada em aminocidos no
proteicos.
10 D um exemplo-tipo de constituintes da saliva de insectos fitfagos que podem elicitar
respostas de defesa das plantas contra os mesmos.
11 Que resposta imediata dada pelas clulas das plantas quando os receptores
especficos destas detectam conjugados de aminocidos com cidos gordos da saliva do
insecto?
12 Indique um metabolito primrio precursor do cido jasmnico bem como os
compartimentos celulares onde decorre a biossntese deste elicitador.
13 Que requisito bioqumico precede a percepo e transduo do sinal desencadeado
pelo cido jasmnico?
14 De forma sucinta, indique o mecanismo de aco desencadeado pelo cido jasmnico
enquanto mensageiro secundrio dos mecanismos de defesa das plantas.
15 Indique, de forma sucinta, os seguintes passos do mecanismo de induo da defesa
sistmica no tomateiro, por insectos fitfagos:
15.1 nas clulas do parnquima flomico das folhas atacadas pelo insecto
15.2 nas clulas de companhia do floema das folhas atacadas pelo insecto
15.3 nos elementos crivosos do floema
15.4 Nas clulas do mesfilo de outras folhas da planta atacada
16 Traduza estes conceitos:
16.1 patognicos necrotrficos
16.2 patognicos biotrficos
16.3 patognicos hemibiotrficos
17 Indique uma classe de molculas que possam ser parte das defesas constitutivas contra
organismos patognicos referindo o seu modo de aco genrico.
18 Em que consiste basicamente a resposta hipersensitiva a um ataque por um fungo
patognico?
19 Descreva, muito sucintamente o mecanismo bioqumico da resposta hipersensitiva
20 Em presena de organismos patognicos, as plantas resistentes, geralmente
desencadeiam duas linhas de resistncia: (a) - a primeira linha de resistncia; (b) um segundo
sistema de resistncia . De forma sucinta, caracterize:
20. 1- (a) - a primeira linha de resistncia
20.2 - (b) o segundo sistema de resistncia
21 Como define o conceito de resistncia sistmica adquirida?
22 Que tipos de molculas e respectivas aces esto relacionadas com a resistncia
sistmica adquirida?
23 - Como define o conceito de resistncia sistmica induzida?
24 Descreva, de forma sucinta, o processo subjacente resistncia sistmica induzida
END
How R and Avr gene products activate plant defense
responses is not yet understood

Each R gene product is thought to possess two functions: recognition of
the corresponding Avr-derived signal, and activation of downstream
signaling pathways to trigger the complex defense response.

The Pto kinase (Table 21.2) confers resistance to strains of Pseudomonas
bacteria that express avrPto, a protein the bacterium delivers directly into
plant cell cytoplasm by type III secretion. There is evidence that Pto kinase
may interact directly with the avrPto signal.

The Pto gene belongs to a linked multigene family and shows 87% sequence
similarity to a second kinase gene called Fen, the product of which confers
sensitivity to the insecticide Fenthion, resulting in an HR type of cell death
(Figure 21.28).
Figure 21.28 Genomic organization of the Pto gene family in tomato (A) and the biological
phenotypes conferred by three of the genes in the cluster (B,C). The Pto gene confers resistance
to Pseudomonas syringae pv. Tomato bacteria expressing the avirulence gene avrPto (B).The Prf
gene is required for both the Fen- and Pto-mediated responses; the Fen gene confers sensitivity
to the insecticide Fenthin (C).
The avrPto/Pto pathway, however, is still incomplete. The
Prf gene, which is located within the Pto gene cluster (see
Fig. 21.28), is required for both Pto- and Fen-specified
responses. Pfr encodes also a LZ-NBS-LRR protein, for
which the molecular function in disease resistance is still
unknown. Conceivably, Prf guards Pto and recognizes
when Pto interacts with Avr-Pto to activate defense.

The cytoplasmic TIR-NBS-LRR class of R proteins can
confer resistance to biotrophic fungi, bacteria, and viruses.





LZ (Leucine Zipper); NBS (Nucleotide Binding Site) ; LRR (Leucine-Rich Repeat)
Several proteins that can interact with Pto kinase have been itentified,
including proteins with homology to transcription factors, (Pti1, Pti2,
Pti3, Pti4, Pti5, Pti6).

Overexpression of Pti kinase in plants carrying the Pto resistance gene
enhances the avrPto-mediated HR. Thus Pto protein appears to activate
several distinct signaling pathways simultaneously (Figure 21.29).
Figure 21.29
Model of the potential Pto-mediated
resistance signaling pathway in tomato.
END