Systemic acquired resistance in Plant Pathology

1. 1

2. SYSTEMIC ACQUIRED
RESISTANCE
2

3.  Plants defend themselves from pathogen infection
through a wide range of mechanisms that can be either
systemic or local, constitutive or inducible.
 Contact with pathogenic and non pathogenic
microorganisms trigger a wide range of defence
mechanisms in plants.
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4. 4
SYSTEMIC PLANT RESPONSES:
Type of systemic response induced, however, is determined
by the identity of the attacking organism.
A)Systemic acquired resistance.
B)Systemic proteinase inhibitor/wound response.
C)Induced systemic resistance (by PGPR).

5. 5
(A) Viruses, fungi, bacteria activate systemically a specific
subset of defence responses in a phenomenon known
as systemic acquired resistance (SAR).
o In which local necrosis formation at the initial site of
pathogen invasion triggers a local increase in salicylic acid
(SA) accumulation i.e the formation of a phloem-mobile
signal.
o Subsequently, in distal plant tissue, SA concentrations
increase and volatile methyl-Salicylate (MeSA) is released.
Together, these signals induce the synthesis of various
pathogenesis related (PR) proteins in the noninvaded
parts of the plant.

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(B) In the systemic PI/wound response activated by chewing
insects, the initial tissue damage causes a transient
increase in the synthesis of ethylene and jasmonic acid
(JA).
o Volatile methyl jasmonate (MeJA) and another phloem-
mobile signal called systemin then activate the systemic
responses, which include the accumulation of PIs and other
systemic wound response proteins (SWRPs).
o Root attacking nematodes appear to induce a mixture of
both the SAR and systemic Pathogen induced/wound
responses.

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8. 8
(C) Induced systemic resistance (ISR) caused by soil
inhabiting nonpathogenic rhizobacteria
colonizing plant roots.
o ISR requires both JA and ethylene- mediated
signaling to induce protective defense responses
in the distant leaf tissue.
o This form of defense does not involve the
accumulation of neither pathogenesis-related
proteins nor SA.

9.  Two main mechanisms have been recognized .
Systemic Acquired Resistance
Induced Systemic Resistance
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10. 10

11. SYSTEMIC ACQUIRED RESISTANCE
oSystemic acquired resistance (SAR) refers to a distinct
signal transduction process, that plays an important
role in the ability of plants to defend themselves
against pathogens.
o After the formation of a necrotic lesion, either as a part of
hypersensitive response (HR) or as a symptom of disease,
the SAR process is activated.
oSAR activation results in the development of a broad-
spectrum, systemic resistance.
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13. HISTORICAL DEVELOPMENT
 The natural phenomenon of resistance development in
response to pathogen infection was first recognized in
1901 by Ray & Beauverie, both who worked with
Botrytis cinerea.
o In 1933, Chester- he first found the concept of SAR and
reviewed research on “the problem of acquired
physiological immunity in plants” and determined that
this phenomenon may play an important role in the
preservation of plants in nature
13

14.  In 1964, Hecht & Bateman showed that inoculation
of tobacco leaves with Thielaviopsis basicola lead
to necrosis and subsequent systemic resistance to
other pathogens, TMV, and TNV.
 Accoring to Smedegaard-Peterson et al both
virulent and avirulent races of
Erysiphe graminis f. sp. hordei could induce
resistance in barley towards virulent strains of this
fungus.
14

15. MECHANISM OF SAR MAINTENANCE
 SAR can be distinguished from other disease
resistance responses by both the spectrum of
pathogen protection and the associated changes in
gene expression.
 In 1970,Van Loon and Gianinazzi et al showed that
viral infection of tobacco induced the accumulation of
a distinct set of proteins , called pathogenesis-related
proteins (PR proteins).
 Van Loon & Antoniw demonstrated that a subset of
PR proteins (acidic-extracellular forms) accumulated
during the onset of resistance. 15

16.  In 1991, steady-state mRNA levels from at least
nine families of genes were shown to be
coordinatedly induced in uninfected leaves of
inoculated plants; these gene families are now
known as SAR genes.
 SAR gene products have direct antimicrobial
activity or are closely related to classes of
antimicrobial proteins.
16

17.  SAR genes include ß-1,3-glucanases, chitinases,
cysteine-rich proteins, which lack known biochemical
function, but have in vitro activity against
Phytophthora infestans.
 However, all the ß-1,3-glucanases should not be
considered to be associated with SAR, rather a gene –
specific subset consisting of mostly acidic isoforms are
precisely correlated with SAR.
17

18. o In tobacco, SAR activation results in a significant
reduction of disease symptoms caused by
the fungi Phytophthora parasitica, Cercospora
nicotianae,and Peronospora tabacina,
the viruses tobacco mosaic virus (TMV) and
tobacco necrosis virus (TNV), and
the bacteria Pseudomonas syringae pv tabaci and
Erwinia carotovora.
18

19. COMPONENTS OF SAR
 Basically SAR has three main components
1. Accumulation of pathogenesis related (PR)
proteins (termed SAR proteins)
2. Lignification and cross-linking of the cell walls of
tissues distant from the HR
3. Phenomenon of conditioning
19

20.  Pathogeneis Related proteins:
 PR-proteins have been defined as plant proteins
that accumulate after pathogen attack or related
situations.
 The role of PRs in defence can be addressed using
transgenic plants expressing the corresponding
genes.
 In tobacco, over expression of PR-1 significantly
increase resistance against infection by
Peronospora tabacina and Phytophthora parasatica
var. nicotianae.
20

21.  Lignification and other Structural Barriers:
 The process of lignification is observed in nonhost
resistance and SAR.
 Lignification may contribute resistance in many
different ways.
 Incorporation of lignin into plant cell wall strengthen
it mechanically and make it more resistant to
degradation by enzymes secreted by an invading
pathogen. 21

22.  Lignin precursors themselves might exert a toxic
effect on pathogens or by binding to fungal cell wall,
make them more rigid and impermeable, thus,
hindering further growth or uptake of water and
nutrients.
 Conditioning/Sensitizing:
 When plants are pretreated with a necrotizing
pathogen or a synthetic inducer of SAR, the
systemically protected leaves react more rapidly
and more efficiently to infection with a virulent
pathogen. This phenomenon is known as
conditioning or sensitizing. 22

23.  The biochemical changes occurring in a sensitized
plant usually become apparent at the moment of
challenge infection.
 At the cellular level, there appears to be a shift
towards a greater sensitivity not only to pathogens
but also biotic and abiotic elicitors.
 Although the phenomenon has long been known,
it is a known effect, whereby, a pathogen
attempting penetration of SAR tissue meets
a more efficient and faster response than it
does on non-SAR tissue. The response may also
occur at lower limits of elicitors.
23

24. REACTIVE OXYGEN SPECIES
 Alvarez et al. had found ROS in Arabidiopsis.
 H2O2 accumulates in small groups of cells in
uninoculated leaves of Arabidiopsis after infection
with an avirulent strain of P. syringae.
 These microbursts occur within two hours after an
initial oxidative burst in the inoculated tissue and
are followed by the formation of microscopic HR
lesions.
 Using catalase to scavange H2O2, or DPI
(diphenylene iodonium) to inhibit the NADPH
oxidase, it was demonstrated that both the primary
and secondary oxidative bursts are required for the
onset of SAR. 24

25. ELICITORS
 Molecules produced by a pathogen that induce a
defence response by the host.
 Innate immunity in plants relies on the rapid
evolution of membrane-bound or cytosolic disease
resistance proteins (R proteins) , which perceive
pathogen protein signatures known as 'elicitors’.
25

26.  Recognition of the elicitors by the host plant
activates a cascade of biochemical reactions in the
attacked and surrounding plant cells and
lead to new or altered cell functions and to new or
greatly activated defence-related compounds.
26

27.  In compatible host–pathogen interactions, the
elicitor escapes recognition by an R protein, leading
to the development of disease.
 In incompatible interactions, the elicitor is
recognized by an R protein, triggering a cascade of
defence reactions that culminate in a form of
programmed cell death (PCD) or apoptopsis, which
is a 'hypersensitive response’ (HR).
27

28.  Typically, HR includes signal transduction , PCD,
increase activation of defence related genes (e.g.,
synthesis if phytoalexins, salicylic acid, and
antimicrobial proteins), and a distant induction of
general defence mechanisms that serve to protect
the plant i.e.,Systemic Acquired Resistance.
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Effector
Virus
Fungi
Bacteria
Pathogen cell
surface
Plant cell
surface
Cell
recognition
Elicitor
receptor site
Elicitors
Signal
transduction
pathways
Defence
genes
Gene
produ
cts
PR
proteins
SAR
Phyto
alexins Chitina
ses
HR
Primary
immune
Response
of
Plants.

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Hypersensitive response and systemic
acquired resistance.Tobacco mosaic
virus infection of tobacco cultivars carrying
a disease resistance gene (e.g. N gene)
lead to the HR and subsequent
establishment of SAR. The HR is
characterized by host cell death and
necrosis at the site of infection (left).
Several days after the primary
infection, SAR is induced throughout the
plant. As a result, secondary infections of
the plant with the same virus or other
unrelated pathogens lead to much smaller
lesions or weaker symptoms (right). The
leaves are shown four days after viral
infection.

32. ROLE OF SALICYLIC ACID IN SAR
o Salicylic acid (SA) plays a key role in both SAR
signaling and disease resistance. Initially, the level
of SA was found to increase by several hundred-
fold in tobacco or cucumber after pathogen
infection, and this increase was shown to correlate
with SAR.
32

33.  The involvement of SA in signaling SAR has been
clearly demonstrated in experiments with
transgenic tobacco that express the salicylate
hydroxylase gene (nahG) from
Pseudomonas putida. This enzyme catalyzes the
conversion of SA to catechol, which has no SAR
inducing activity.
 The nahG expressing plants do not accumulate SA
in response to pathogen infection and they show no
SAR gene or resistance expression in the distal
portions of the plant.
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34. 34
o Even though SA is not likely to be the translocated
signal that triggers SAR in distal plant organs, it is
essential for SAR signal transduction.
oInoculation of wild-type rootstocks with TMV leads to the
induction of SAR in wild type scions but not in nahG
scions, demonstrating that the induction of SAR in
systemic tissues is SA dependent.
oThese findings indicate that SA is an essential signal in
SAR and that it is required downstream of the long
distance signal.

35. BIOSYNTHESIS OF SALICYLIC ACID
 The biosynthetic pathway of SA in plants begin with
the conversion of phenylalanine to trans-cinnamic
acid (t-CA) catalyzed by phenylalanine ammonia
lyase (PAL).
 The conversion of t-CA into SA has been proposed
to proceed via chain shortening to produce benzoic
acid (BA), followed by hydroxylation at the C-2
position to derive SA .
35

36. o The next step is likely to be catalyzed by a
cytochrome P450 mono oxygenase, called benzoic
acid 2-hydroxylase (BA2H), the activity of which is
induced by either pathogen infection or exogenous
BA application.
o The mechanism of BA production from t-CA is
unknown, but it may occur in a manner of
β-oxidation of fatty acids (or) a nonoxidative
mechanism.
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37. 37
phenylalanine
Cinnamic acid

38. MODE OF ACTION OF SA
o The mechanism by which SA induces SAR is unknown;
however, it has been proposed that H2O2 acts as a
second messenger of SA in SAR signaling.
 Biochemical screens for SA-binding proteins, by Klessig
et al. resulted in the identification of multiple enzymes,
such as catalase, ascorbate peroxidase, the E2
subunit of α-ketoglutarate dehydrogenase, and
glutathione S-transferases, showed that their enzymatic
activities are inhibited upon binding to SA.
38

39. o SA decreases the expression of ascorbate
peroxidase (APX) & catalase (CAT) genes leading
to substantial increase in H2O2 .
o The later causes induction of PR-1 gene
expression and induce SAR.
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Fig. 1. SA decreases the
expression of ascorbate
peroxidase (APX) & catalase
(CAT) genes leading to substantial
increase in H2O2 which is crucial
for the activation of local acquired
resistance (LAR) &
systemic acquired resistance
(SAR).

41. NITRIC OXIDE (NO) AS SIGNAL MOLECULE:
NO is involved in multiple regulatory processes in plants.
o In plants, NO and SA appear to function in a positive
feedback loop.
o NO donors induce SA accumulation, and NO signaling
in defence requires SA.
o Supporting this hypothesis nahG expression suppressed
NO-inducible local and systemic resistance in TMV-
infected tobacco, where as SA –induced SAR was
promised by an NO scavenger or inhibitors of NO
synthesis.
 NO is involved locally in the induction of cell death (HR)
in conjunction with SA, H2O2, and ethylene.
41

42. LIPID-TRANSFER PROTEIN
o The dir1 (defective in induced resistance 1) mutant
was discovered in a labor-intensive genetic screen
designed specifically to identify SAR Signals.
o In contrast to other SAR-deficient mutants, dir1 has a
normal local defence response but is compromised in
SAR.
o Even though petiole exudates collected from dir1
lack the SAR-inducing activity, the mutant is
competent in responding to induction by the petiole
exudates collected from induced wild-type plants.
42

43. o This suggests that DIR1, which encodes a putative
lipid-transfer protein, is probably involved in the
synthesis or transport of a lipid molecule, which is a
mobile signal for SAR.
o Recently a tobacco SA-binding protein SABP2 a lipase
was discovered and was found that, silencing of
SABP2 gene diminishes both SRI and ISR.
43

44. SAR ACTIVATION WITH CHEMICALS
For the chemical to be considered as an activator of
SAR, it should exhibit these characteristics.
 First, the compound or its significant metabolites
should not exhibit direct microbial activity.
 Second, it should induce resistance against the
same spectrum of pathogens as in biologically
activated SAR.
 Third, it should induce the expression of the same
marker gene as evident in pathogen-activated SAR. 44

45.  Low molecular weight molecules are also capable
for SAR induction in plants.
 The use of chemicals to activate SAR provides
novel alternatives for disease control in agronomic
systems as well as tools for the elucidation of the
SAR signal transduction cascade.
45

46. Natural organic compounds:
 SA is the only plant derived substance that has been
demonstrated to be an inducer of SAR.
 The chemicals 2, 6-dichloroisonicotinic acid and its
methyl ester were the first synthetic compounds shown
to activate SAR, providing broad spectrum disease
resistance.
 Synthetic chemical benzo-1,2,3-thiadiazole-7-carbothioic
acid S methyl ester (BTH) was demonstrated to be a
potent SAR activator that showed protection in the field
against a broad spectrum of diseases in a variety of
crops. 46

47. Inorganic compounds:
 Phosphate salts induce SAR in cucumber bean and
maize.
 Calcium sequestration at the site of application by
phosphates is thought to generate endogenous
SAR signal.
47

48. Synthetic compounds:
 Probenazole is commercial product used against
rice blast. It has a little effect on rice prior to
inoculation, but potentiates defence responses
upon attack by Magnaporthe oryzae.
 Benzo-(1, 2, 3)-thiazole-7-carbothionic acid S-
methyl ester (BTH) is an inducer of SAR in a
number of plants including wheat, rice and tobacco.
Its tolerance and efficacy in these crops have
warranted its commercialization.
48

49.  2,6-dichloroisonicotinic acid induces changes in rice
and barley where treated plants exhibit
phenocopies of genetically determined resistance
against Erysiphe graminis and
Magnaporthe oryzae.
49

50. 50
Salicylic acid 2,6-dichloroisonicotinic acid
(INA)
Benzo-(1,2,3)-thiodiazole-7-
carbothioic acid S-methyl ester
(BTH).

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52. SAR COMPROMISED MUTANTS
o Delaney et al., 1994 identified and characterized six
allelic recessive mutants named nim.
o These are not responsive to exogenous application
of SA.
o Cao et al., have also isolated and described
recessive Arabidopsis mutant called npr1 (non
expresser of PR genes), which exhibits
compromised activation of SAR npr1 and are allelic
to nim1 (non inducible immunity).
o In these both SAR signaling is blocked before SAR
gene expression but subsequent to SA
accumulation. 52

53.  nim 1 plants infected with the avirulent pathogen
Pseudomonas syringae DC 3000 harboring the
cloned avr Rpt 2 .
 nim 1 plants were shown to accumulate both free
and glucose conjugated SA levels.
 nim 1 plants are able to accumulate SA in response
to pathogen infection but appear to be defective in
SA perception or in subsequent SA sensing events.
 The nim1 and npr1 mutations define genes that act
before SAR gene expression substantiated by the
lack of PR-1 induction. 53

54. FITNESS COSTS OF SAR
 Phenotypes of many mutants show constitutive PR
gene expression, accumulation of SA and
resistance to pathogen, but have reduced plant
size, loss of apical dominance, curly leaves and
decreased fertility.
 Treatment with BTH in plants grown hydroponically,
shown a reduction in biomass and number of ears
and grains in wheat.
 Cost of resistance could be due to the allocation of
the plant’s resources to constitutive PR protein
production.
 Thus constitutive expression of SAR in uninfected
plants is found to be detrimental. 54

55. INDUCED SYSTEMIC RESISTANCE
 Roots of some plants are colonized by specific
strains of non-pathogenic fluorescent
Pseudomonas spp. and they develop
phenotypically similar form of protection that is
called rhizobacteria-mediated induced systemic
resistance (ISR).
 ISR is controlled by signaling pathway in which
jasmonic acid and ethylene play key roles.
 Rhizobacteria are present in the rhizoplane and
certain strains of it are referred to as plant growth
promoting rhizobacteria (PGPR) because, they can
stimulate growth of the plants. 55

56.  Rhizobacteria mediated ISR in Arabidiopsis
colonization of Arabidiopsis roots by ISR inducing
Pseudomonas fluorescens WCS417r bacteria
protects the plant against different types of
pathogens including the bacterial leaf pathogens
Pseudomaonas syringe pv. tomato and
Xanthomonas campestris pv. armarociol, the fungal
root pathogen Alternaria brassicicola and oomycete
leaf pathogen Peronospora parasitica.
 Protection against these pathogens is typically
manifested as reduction in disease symptoms and
inhibition of pathogen growth. 56

57. 57

58. SYNTHESIS OF JASMONIC ACID
 Jasmonic acid (JA), is a growth inhibitor, senescence-
promoting substance.
 Jasmonic acid synthesized from linolenic acid via the
octadecanoic pathway.
 JA is synthesized from alpha-linolenic acid, which is a
C18 poly-unsaturated fatty acid.
 It is synthesized from plant membranes by enzymes
similar to lipase.
 Key JA biosynthetic enzymes include lipoxygenase,
allene oxide synthase, and allene oxide cyclase.
58

59.  JA induces vegetative storage proteins, osmotin,
thionin (antifungal) and defensin.
 Induces protease inhibitors to control pests.
 JA and ethylene induce PR-3, PR-4, PDF1.2,
chitinases (CHI-B).
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OCCURS IN
CHLOROPLAST
OCCURS IN
PEROXISOME
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62. ISR SIGNAL TRANSDUCTION PATHWAY
 The role of SA in ISR was studied in SA non
accumulating Arabidiopsis NahG plants. In contrast to
pathogen induced SAR, Pseudomonas fluorescens
WCS 417r-mediated signal ISR against Psuedomonas
syringae pv. tomato was normally expressed in these
plants. (Van Wee et al., 1997;Pieterse et al., 1996).
 SA induction deficient mutants Sid 1-1 and Sid 2-1
expressed normal levels of ISR.
62

63.  Determination of SA levels in ISR expressing
Arabidopsis plants revealed that in contrast to SAR,
ISR is not associated with increased accumulation
of SA (Pieterse et al., 2000).
 This has led to the conclusion that
Pseudomonas fluorescens WCS417r mediated ISR
is an SA independent resistance response, and that
ISR are regulated by distant signaling pathways.
63

64.  Wounded leaves produce an 18-amino acid peptide
called systemin from carboxyl terminal of prosystemin
(200 AA precursor) in plant and it elicits production of
JA in companion cell-seive element complex.
 JA moves through out the plant in the phloem.
Covalently modified JA (JA-x) play important role in
systemic signaling.
 Signal is recognized at the target cell e.g. mesophyll
(leaf).
 Jasmonic acid turns on defence related genes (genes
for proteinase inhibitor) in target cells.
64

65. 65

66. CROSSTALK BETWEEN SA AND JA
o Both synergistic and antagonistic interactions
between SA and JA have been reported,
suggesting that the interaction is either
concentration dependent or tissue specific.
 Application of low concentrations of SA and JA
resulted in synergistic expression of both the SA
target PR1 and the JA marker PDF1.2 and Thi1.2.
66

67.  In contrast, higher concentrations of SA and JA
have antagonistic effects on expression of the
same genes.
 It is generally believed that this antagonism
between SA and JA allows plants to prioritize
defence against either biotrophic or necrotrophic
pathogens.
o Host plants rely on SA-mediated defence against
biotrophic pathogens, whereas JA-mediated
signaling participates in defence against
necrotrophic pathogens.
67

68. 68

69. COMPARISON OF ISR AND SAR:
 Both ISR and SAR are dissimilar with respect to
their induction, signaling pathways and their co-
ordination, level of defense elicited in host plant and
protein expression.
o ISR is induced by jasmonic acid and ethylene
molecules coordinated signaling pathway; proteins
other than pathogenesis related are too expressed.
o Level of defense generated is lesser and also
dependent on host plant genotype and presence of
non-pathogenic bacteria.
69

70. o SAR is induced by salicylic acid. SAR and PR
proteins are induced together.
 The defense level produced by SAR is high, long
lasting, broad spectrum and independent of plant
genotype but requires necrotic pathogens or
presence of inducer analogs.
 It has been indicated that ISR and SAR show
phenotypic similarity and together can produce
protection of higher level than they produce alone.
70

71. ARTICLE:
 The effects of riboflavin on defence responses and
secondary metabolism in tobacco (Nicotiana tabacum
cv. NC89) cell suspensions and the effects of
protecting tobacco seedlings against Phytophthora
parasitica var.nicotianae and Ralstonia solanacearum
were investigated.
 Defence responses elicited by riboflavin in tobacco
cells induced an oxidative burst, alkalization of the
extracellular medium, expression of 4 defence-related
genes and 2 phenolic compounds.
 When applied to tobacco plants riboflavin treatment
gives protection.
71

72.  Finally conclude that riboflavin can both induce a
series of defence responses and secondary
metabolism in cell suspensions and protect tobacco
against P. parasitica and
R. solanacearum
o Exogenously applied sphinganine produced similar
effects on these markers.
 This suggests that fumonisin production contributes
to the colonization of this necrotroph by activating
the SA pathway and inducing cell death
72

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74. REFERENCES:
 Agrios,G.N., (2005). Plant Pathology; Fifth Edition;
341-348.
 Helmut Kessmann et al., (1994). Induction of
Systemic Acquired Disease Resistance in Plants by
Chemicals, Annual Review of Phytopathology.
32:439-459.
 Michal Shoresh, Gary E.Harman, and Fatemeh
Mastouri.,(2010). Induced Systemic Resistance and
Plant Responses to Fungal Biocontrol Agents,
Annual Review of Phytopathology,48:21-43.
74

75.  Rajan Katoch.,(2010). Biochemistry and Molecular
Biology of Plant Disease Resistance; First Edition;
73-89.
 Raskin I.,(1992).Role of salicylic acid in plants,
Annual Review of Plant Physiology and Plant
Molecular Biology,43:439-463.
 W.E. Durrant and X. Dong.,(2004). Systemic
Acquired Resistance, Annual Review of
Phytopathology,42:185-209.
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