Plant Signaling & Behavior

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SA-ABA antagonism in defense responses

Wolfgang Moeder, Huoi Ung, Stephen Mosher & Keiko Yoshioka

To cite this article: Wolfgang Moeder, Huoi Ung, Stephen Mosher & Keiko Yoshioka (2010)
SA-ABA antagonism in defense responses, Plant Signaling & Behavior, 5:10, 1231-1233, DOI:
10.4161/psb.5.10.12836

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Published online: 01 Oct 2010.

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 article addendum
Plant Signaling & Behavior 5:10, 1231-1233; October 2010; © 2010 Landes Bioscience
SA-ABA antagonism in defense responses
Wolfgang Moeder,1,2 Huoi Ung,1 Stephen Mosher1 and Keiko Yoshioka1,2,*
1
Department of Cell and Systems Biology; 2Center for the Analysis of Genome Evolution and Function (CAGEF); University of Toronto;
Toronto, ON Canada
Key words: salicylic acid, SA, abscisic
acid, ABA, lesion mimic mutant, envi-
ronmental effects, humidity, drought
stress, water loss
Submitted: 06/28/10
Accepted: 06/29/10
Previously published online:
www.landesbioscience.com/journals/psb/
article/12836
DOI: 10.4161/psb.5.10.12836
*Correspondence to: Keiko Yoshioka;
Email: keiko.yoshioka@utoronto.ca
Addendum to: Mosher S, Moeder W, Nishimura
N, Jikumaru Y, Joo SH, Urquhart W, et al. The
lesion mimic mutant cpr22 shows alterations in
abscisic acid signaling and abscisic acid insensi-
tivity in a salicylic acid-dependent manner. Plant
Physiol 2010; 152:1901–13; PMID: 20164209; DOI:
10.1104/pp.109.152603.
www.landesbioscience.com Plant Signaling & Behavior 1231
U ntil recently, phytohormones were
mostly studied separately. However,
recent studies have suggested that the
signaling pathways involved are highly
interconnected. We recently reported
the antagonistic effects of salicylic acid
(SA) and abscisic acid (ABA) in the
lesion mimic mutants, cpr22 and ssi4.
After shifting these mutants from high
humidity, where the lesion mimic phe-
notype is suppressed to permissive low
humidity condition, both SA and ABA
pathways were upregulated. However,
the increased levels of SA were able to
block downstream ABA responses even
though ABA signaling genes and endog-
enous ABA were elevated. Furthermore,
these lesion mimic mutants displayed a
partial ABA insensitivity with respect
to germination, guard cell opening and
water loss. This increased water loss in
detached mutant plants could also be
mimicked by treating wild type plants
with SA. An active SA analog, 5-chloro-
salicylic acid also induced enhanced
water loss, while an inactive analog,
4-hydroxy-benzoic acid, did not. Here,
we report that the biological analogs
of SA, the systemic acquired resistance
(SAR) activators, BTH [benzo-(1,2,3)-
thiadiazole-7-carbothioic acid S-methyl
ester) and BIT (1,2-benzisothiazol-
3(2H)-one1,1-dioxide], did not have the
same effect as SA, suggesting that SA
may have additional roles to defense and
that SAR activators may not mimic all
SA effects.
We recently reported an intriguing antago-
nism between salicylic acid (SA)—abscisic
acid (ABA) in the Arabidopsis lesion
mimic mutant, cpr22.1 SA is a well known
article addendum
signaling molecule for defense responses in
a variety of plant species. Generally, there
are two categories of defense responses
in plants: (1) basal immunity, which is
activated by molecules that are conserved
among many pathogens (PAMPS; patho-
gen-associated molecular patterns) and is
also known as PAMP-triggered immunity
(PTI) and (2) a stronger defense response,
which is mediated by resistance (R) genes
that can interact with particular effec-
tors from the pathogen or can recognize
effector-induced modifications of plant
proteins, also known as effector-triggered
immunity (ETI).2 One important defense
mechanism activated by R gene-mediated
pathogen recognition is the hypersensitive
response (HR), which is characterized by
apoptosis-like cell death at and around the
site of pathogen entry.3 HR development
is usually accompanied by an increase
in SA and the accumulation of defense-
related proteins such as the pathogenesis-
related (PR) proteins.4 It has been shown
that SA is required for the establishment
of both local and systemic resistance
against various biotrophic and hemi-bio-
trophic pathogens using the SA biosynthe-
sis mutant, SA induction-deficient2 (sid2),
as well as NahG transgenic plants that can
not accumulate SA.5,6
The Arabidopsis mutant, constitutive
expresser of PR genes22 (cpr22), was iso-
lated as a mutant exhibiting elevated levels
of SA and constitutive expression of PR
genes without pathogen infection.7,8 This
mutant is also resistant to the virulent bac-
terial pathogen, Pseudomonas syringae and
the oomycete pathogen, Hyaloperonospora
arabidopsidis. In addition to this height-
ened resistance phenotype, cpr22 also
shows HR-like spontaneous cell death.
 lesion mimic phenotype was suppressed
while after the shift, HR cell death
started to become visible. Interestingly,
in shifted plants, we observed an altera-
tion in the expression of ABA-related
genes in addition to an upregulation of
SA/defense-related genes.1 We found that
ABA biosynthetic and upstream signal-
ing genes, such as 9-cis-epoxycarotenoid
dioxygenase3 (NCED3), aldehyde oxidase3
(AAO3) and ABI1, were strongly upregu-
lated while far-downstream genes, such as
RD22, RD29A, KIN1/2 and ERD3, were
surprisingly downregulated. This indi-
cates two intriguing phenomena: (1) an
over-activation of the ABA signaling path-
way and (2) a suppression in the down-
stream of the ABA pathway, preventing
the expression of downstream genes in
both mutants. In accordance with (1),
the measurement of endogenous ABA
levels confirmed a temporal increase of
endogenous ABA in both mutants after
humidity shift.1 The alteration of the
Figure 1. ABA signaling genes that show alteration in cpr22 and ssi4 after humidity shift. Red expression pattern of ABA-related signal-
arrows (↑) indicate genes that were induced more than two-fold in these mutants compared to ing is summarized in Figure 1. A likely
wild-type plants. Blue arrows (↓) indicate genes that were suppressed more than two-fold in
these mutants. The grey arrow (↔) indicates genes that did not show any significant difference in
explanation for this discrepancy is that
mutants and wild-type plants. The figure was created based on the ABA signaling model created firstly, ABA signaling was suppressed at
by Shinozaki et al. (2003).13 Original microarray data are available from the Bio-Array Resource for the level of ABA-related transcription fac-
Plant Functional Genomics (BAR) web site under Project 50 of BAR’s project browser (bar.utoronto. tors, such as MYC2 and AREB/ABF that
ca/affydb/cgi-bin/affy_db_proj_browser.cgi). bind to various ABA-responsive elements.
Consequently, the genes that are under the
Therefore, cpr22 is categorized as a lesion used two environmentally sensitive lesion control of these elements, such as RD29
mimic mutant.9 mimic mutants, cpr22 and ssi4 (suppres- and RD22, are suppressed. This suppres-
It has been reported that some lesion sor of salicylic acid insensitive4).7,11 When sion of far-downstream signaling likely
mimic mutants are environmentally sen- analyzing the effect of environmental prevents the plants from responding prop-
sitive, i.e., their resistance phenotypes conditions on pathogenesis, difficulties erly to abiotic stresses (i.e., acute humid-
are conditional.10 For instance, under arise because we cannot conclude whether ity change) and enhances stress damages.
high humidity conditions, such as on the plant, the microorganism, or both A feedback system may then be activated
agar plates or when grown at high tem- are being affected by environmental fac- to compensate for this suppression by fur-
perature, both the spontaneous HR and tors. Furthermore, it has been reported ther over-activating some up-stream sig-
the enhanced pathogen resistance is that some effectors of microbes actively naling components (Fig. 1). Interestingly,
suppressed.7,11 On the other hand, rela- manipulate plant hormone signaling.12 we have shown that this alteration in ABA
tively low humidity or cold temperature Therefore, it is generally difficult to study signaling is due to the accumulation of SA
enhances their SA-related phenotypes, the environmental effect on the plant by conducting humidity shift experiments
including HR-like cell death.1 Similar defense response using pathogens. The using a cpr22/NahG double mutant.1
environmental effects were also reported use of environmentally sensitive mutants Thus, we can speculate that SA accumu-
for the response of wild-type R genes, sug- that show activation of defense responses lation antagonizes ABA signaling after or
gesting that there is a universal factor(s) in without pathogen infection, such as cpr22 around the above-mentioned transcription
defense signaling that is environmentally and ssi4, can eliminate this obstacle. factors.
sensitive.10 However, the precise molecular Therefore, we conducted genome-wide The phytohormone, ABA, controls
mechanism of this environmental sensitiv- expression analysis with both mutants, various environmental (abiotic) stress
ity is not clear, since such a factor(s) has comparing plants grown at ∼95% relative responses, including drought, salinity
not yet been identified. humidity with plants that were shifted and temperature stress.13 Since we have
To characterize this phenomenon and from 95% humidity to 65% humidity. observed an alteration in ABA signaling
find the component(s) involved, we have Under high humidity conditions, the in cpr22 and ssi4, we further characterized

1232 Plant Signaling & Behavior Volume 5 Issue 10

ABA-related phenotypes, such as ABA
responsiveness of guard cell closure and
germination and water-loss phenotypes
in both mutants. As we expected, both
mutants showed a partial deficiency in
their ABA responsiveness. One strik-
ing phenomenon we observed was the
enhanced water loss phenotype in these
mutants.1 Both mutants, as well as
another environmentally sensitive lesion
mimic mutant, copine1/bonzai1 (cpn1/
bon1) 14 showed this enhanced water loss
phenotype. We further demonstrated
that this phenotype is due to the action
of SA, since wild type plants treated with
SA also displayed enhanced water loss.1
We have also tested an active SA analog,
5-chloro-salicylic acid, which similarly to
SA induces PR genes. This compound also
induced the same water loss phenotype in
wild-type plants (data not shown) while
an inactive SA analog, 4-hydroxy-benzoic
acid, did not. As far as we know, this is the
first report that clearly demonstrates that
SA enhances water loss in plants.
This observation raised the question
of whether other types of biological ana-
logs of SA for pathogen defense responses
can also induce the same water loss
effect. It has been reported that synthetic
chemicals, such as BTH, INA (methyl-
2,6-dichloroisonicotinic acid) and BIT
(the active compound of Probenazole),
can induce pathogen resistance responses
similar to SA and they are considered bio-
logical analogs of SA (also known as SAR
activators).15-17 Thus, we tested the effects
of BTH and BIT on wild-type plants.
As shown in Figure 2, both BTH and
BIT do not have the same effect as SA in
inducing elevated water loss in detached
plants. This result was unexpected, since
the structural analog, 5-chloro-salicylic
acid, is active both in terms of pathogen
defense response as well as water loss,
while 4-hydroxybenzoic acid is inactive
in terms of defense and water loss, indicat-
ing a correlation between effects on water
loss and pathogen defense responses.1
However, our new data (Fig. 2) suggests
that SA may have additional roles outside
of defense and that SAR activators may
not mimic all effects of SA. Currently, we
are further investigating the effect of SA
on water loss stress responses in order to
www.landesbioscience.com Plant Signaling & Behavior 1233
Figure 2. Effect of SAR activators on water-loss phenotype. The experiment was conducted as
described before (Mosher et al. 2010)1 using Columbia wild-type plants. All treatment has been
done by soil drenching with 1 mM solutions of SA, BTH and BIT.
understand the role of SA in abiotic stress
responses.
Recently, it is becoming clear that ABA
is also involved in biotic stress responses in
a complex manner.18 On the other hand,
the effect of SA on abiotic stress responses
is also being revealed.1,19 Considering that
plants undergo continuous exposure to
multiple stresses under natural conditions,
the signal transduction pathways for abi-
otic and biotic stress responses have to be
interconnected to allow plants to coordi-
nate and prioritize their reactions for their
survival using limited resources. Indeed,
recent studies about the crosstalk between
these signal cascades are becoming a cen-
tral theme in various research fields. Thus,
we believe our finding regarding the effect
of SA on abiotic stress responses will sig-
nificantly contribute to our understand-
ing of the complexity of crosstalk between
abiotic and biotic stress responses.
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