THE ROLE OF

SALICYLIC ACID
IN SAR
Theresa Paula A. dela Rosa
INTRODUCTION
 Salicylic acid (SA) – a phenolic compound
 Salicylates – (general term for SA and it
derrivatives)
 Pharmacological properties have been long valued
 5th century BC by Hippocrates as pain relief during
childbirth
 1828 Johan Buchner- salicyl alcohol glucoside
(salicilin) from willow tree
INTRODUCTION
 Raffaele Piria – salicilin to sugar and acid (SA)
 1859 Hemann Kolbe – synthetic production
 1897 Felix Hoffmann – synthetic derivative acetyl
salicylic acid (ASA), C.F. Gerhardt 1853
 Spontaneous hydrolysis to SA, less GI irritation,
same therapeutic properties
 1899 Bayer Company – Aspirin, most successful
and widely used drug worldwide
INTRODUCTION
 In plants:
 plant-water relationships (Barkosky and Einhellig 1992)
 thermogenesis (Raskin et al. 1987)
 seed germination (Rajou et al. 2006)
 seedling establishment (Alonso-Ramırez et al. 2009)
 cell growth (Vanacker et al. 2001)
 respiration (Norman et al. 2004)
 stomatal responses (Manthe et al. 1992; Lee 1998)
 senescence (Rao et al. 2001, 2002)
 thermotolerance (Clarke et al. 2004)
 and nodulation (Stacey et al. 2006)
INTRODUCTION
 In addition, genetic mutant studies in Arabidopsis
suggest that SA is involved in:
 modulating cell growth (Rate et al. 1999)
 trichome development (Traw and Bergelson 2003)
 leaf senescence (Morris et al. 2000).

 effect on some of these processes may be indirect,

because SA is heavily involved in crosstalk with other
plant hormones (Robert-Seilaniantz et al. 2007; Yuan
et al. 2008; Pieterse et al. 2009)
INTRODUCTION
 SA as a signaling molecule in plant immune response (Vlot et al.
2009)
 plants lack specialized immune cells and immunological memory
 Recognition of pathogen-associated molecular patterns (PAMPs)
results in PAMP-triggered immunity (PTI, formerly called basal
resistance)
 pathogens have evolved effectors to dampen PAMP-triggered
signals
 host plants in turn have evolved resistance (R) proteins to detect
the presence of pathogen effectors and induce effector-triggered
immunity (ETI)
SA METABOLISMS
(BIOSYNTHESIS)
 phenylalanine ammonia lyase (PAL)-mediated
phenylalanine pathway
 isochorismate synthase (ICS) mediated isochorismate
pathway
 Chorismate as the primary metabolite, an
intermediate of plant phenylpropanoid pathway
(downstream of shikimic acid pathway leading to
lignin, flavonoid, anthocyanin, and proanthocyanidin
biosynthesis)
SA METABOLISMS
(BIOSYNTHESIS)
SALICYLIC ACID IN
PATHOGEN DEFENSE
 Study by Ross in 1960 showed convincingly that infection of a
local lesion tobacco host with Tobacco Mosaic Virus (TMV)
conferred substantial local and systemic resistance to a
secondary TMV inoculation (Ryals et al. 1994)
 Ross called this induced whole-plant resistance response
systemic acquired resistance (SAR)
 induction of SAR is accompanied by production of
approximately a dozen families of soluble basic and acidic
proteins called Pathogenesis-Related (PR) proteins; the genes
encoding these proteins have been characterized and most are
induced during SAR.
SYSTEMIC ACQUIRED
RESISTANCE (SAR)
 (SAR) is a "whole-plant" resistance response that occurs following an
earlier localized exposure to a pathogen.
 Plants use pattern-recognition receptors to recognize conserved microbial
signatures. This recognition triggers an immune response. SAR is important
for plants to resist disease, as well as to recover from disease once formed.
 activation of SAR requires the accumulation of endogenous salicylic acid
(SA)
 The pathogen-induced SA signal activates a molecular signal transduction
pathway that is identified by a gene called NIM1, NPR1 or SAI1 (three
names for the same gene) in the model genetic system Arabidopsis
thaliana.
 Similarly, experiments by Gaffney 1993; Vernooji et al. showed that SAR
requires the accumulation of SA for its expression using transgenic tobacco
plants expressing the bacterial salicylate hydroxylase (NahG) gene (NahG
APPLICATION OF SA TO
PLANTS TRIGGERS SAR
 Experiments ny White 1979 - induction of TMV resistance by
infiltration of tobacco leaves with dilute solutions of
acetylsalicylic acid (aspirin) or SA
 also induced production of PR proteins
 application of SA could mimic pathogen-induced SAR

 Shang et al. 2010 – JA and SA, improve plant resistance to RNA

viruses
 Cucumber mosaic virus (CMV), Tobacco mosaic virus (TMV) and Turnip
crinkle virus (TCV) in Arabidopsis, tobacco, tomato and hot pepper.
 The inhibition efficiency to virus replication usually achieved up to 80–
90%.
SA-DEFICIENT PLANTS FAIL TO ACTIVATE
SAR, ARE HYPERSUSCEPTIBLE TO
PATHOGENS, AND COMPROMISED IN
EXPRESSION OF RACE-SPECIFIC
RESISTANCE
 Ryals and colleagues created plants that could not accumulate SA and
assessed them for the ability to activate SAR.
 This was achieved by expressing in tobacco plants the bacterial NahG gene
that encodes salicylate hydroxylase (SH). Salicylate hydroxylase converts
SA into catechol.
 NahG plants were tested for SAR
 inoculating lower leaves with TMV
 later challenging upper leaves with a secondary inoculation of TMV
 NahG and wild type were examined for SA accumulation 7 DAI
 wild type plants,185-fold greater SA levels than untreated plants
 NahG 2-3-fold increase
 Normal tobacco showed a typical SAR response, with
lesions on the challenge leaves about 40% lesser than
the diameter of lesions produced on naive, mock-
inoculated plants that did not receive a primary
inoculation.

 However, NahG tobacco plants showed no evidence of

SAR, with the upper leaf lesions equal in size on NahG
plants that experienced or did not receive a primary
inoculation.
 Delaney et al. 1994
 NahG tobacco plants inoculated with TMV
 accompanied by larger tobacco mosaic virus (TMV) lesions
than wild-type (Xanthi) plants.
 Eventually, TMV lesions expanded from the leaf to the stem
on infected NahG plants.
 Stem dissection showed that all tissues, including the
vascular tissue, had undergone necrosis, which was
associated with the presence of viral RNA as determined by
RNA hybridization experiments.
 Stem necrosis was observed consistently in NahG plants, but
not in Xanthi plants
 Increased disease susceptibility was a general feature of NahG plants
 NahG showed more severe disease symptoms than wild-type plants when
inoculated with Pseudomonas syringae pv. tabaci, Phytophthora parasitica or
Cercospora nicotianae
 Expression of the NahG gene in plants produces a phenotype of enhanced
disease susceptibility and suppression of genetic resistance

 Zabala et al. 2009

 Antagonism between SA and ABA
 that abscisic acid signaling pathways are utilized by the bacterial
phytopathogen Pseudomonas syringae to promote pathogenesis in
Arabidopsis thaliana
 SA is required in order to mount a full innate immune response, while bacterial
effectors act rapidly to activate ABA biosynthesis
 ABA suppresses inducible innate immune responses by down-regulating SA
biosynthesis and SA-mediated defenses
ACCUMULATION OF SA IS NEEDED FOR
RACE SPECIFIC OR GENE-FOR-GENE
RESISTANCE
 Huang and Ma 1992
 Arabidopsis thaliana ecotype Columbia (Col-0) plants can express the NahG gene
 Pseudomonas syringae pv. Tomato (DC3000) is virulent on Col-O plants
 On Col-O plants, DC3000 caused the formation of small, chlorotic spots on
inoculated leaves that were associated with an increase of four to five orders of
magnitude in bacterial titer over 5 days.
 However, when NahG-expressing plants were inoculated with DC3000, the
bacterial titer was 10 to 50 times greater than that seen in non-transgenic
controls; the increased growth was accompanied by severe disease symptoms
 Noco race of Peronospora parasitica causes downy mildew
disease on Arabidopsis ecotype Col-0
 Transgenic Col-O (NahG) plants exhibited much greater susceptibility to
Noco than wild-type Col-0, leading to heavy production of conidia and
oospores

 The Wela race of P. parasitica is not virulent on Col-0 plants,

because of a single R gene that triggers a HR
 Col-0 (NahG) plants supported growth of this fungal isolate
 severe disease symptoms and production of abundant hyphae, conidia,
and oospores.
 By 3 weeks after inoculation with Wela, NahG plants completely
succumbed to the pathogen, which was never seen with this Arabidopsis-
Peronospora interaction.
 Col-O (NahG) plants were also susceptible to another P.
parasitica race called Emwa
 wild-type Col-O plants have genetic resistance, encoded by an
independent R gene.
 These results indicate that expression of NahG suppressed the action of
R genes that confer resistance to both races of P. parasidica.

 The failure of NahG plants to express SAR, their

hypersusceptibility to virulent pathogens, and the breakdown
they display in expression of race-specific resistance, indicates
that SA plays a central role in expression of plant defense.