Plant Growth Regulation 30: 157161, 2000.

2000 Kluwer Academic Publishers. Printed in the Netherlands.

157

Acetyl salicylic acid (Aspirin) and salicylic acid induce multiple stress
tolerance in bean and tomato plants
Tissa Senaratna , Darren Touchell, Eric Bunn & Kingsley Dixon
Kings Park and Botanic Garden, West Perth, WA 6005, Australia
( author for correspondence; e-mail: tissa@kpbg.wa.gov.au)
Received 1 June 1999; accepted 24 August 1999

Key words: stress tolerance, acetyl salicylic acid, salicylic acid, heat, drought, cold

Abstract
The hypothesis that physiologically active concentrations of salicylic acid (SA) and its derivatives can confer
stress tolerance in plants was evaluated using bean (Phaseolus vulgaris L.) and tomato (Lycopersicon esculentum
L.). Plants grown from seeds imbibed in aqueous solutions (0.10.5 mM) of salicylic acid or acetyl salicylic
acid (ASA) displayed enhanced tolerance to heat, chilling and drought stresses. Seedlings acquired similar stress
tolerance when SA or ASA treatments were applied as soil drenches. The fact that seed imbibition with SA or
ASA confers stress tolerance in plants is more consistent with a signaling role of these molecules, leading to the
expression of tolerance rather than a direct effect. Induction of multiple stress tolerance in plants by exogenous
application of SA and its derivatives may have a significant practical application in agriculture, horticulture and
forestry.
Abbreviations: SA salicylic acid; ASA acetyl salicylic acid; SAR systemic acquired resistance

1. Introduction
One of the most important factors that dictates the
distribution of many plant species is their ability
to withstand environmental stress including seasonal
variations in temperature and available moisture.
Plants generally respond to environmental stress by
activating defence mechanisms and adjusting their cellular metabolism [8, 16]. Plants perceive the stress
condition and signal to alter the metabolic flux for
the activation/synthesis of defence mechanisms [16].
Many molecules, for example, calcium, jasmonic acid,
ethylene and salicylic acid have been suggested as signal transducers or messengers [11]. Salicylic acid (SA)
has received much attention after the discovery of its
ability to induce resistance (systemic acquired resistance or SAR) to pathogens [13, 19, 20, 25]. Exogenous application of SA induced pathogenesis-related
gene expression and systemic acquired resistance [2].
Extensive studies have been undertaken to elucidate the molecular biology of SA induced SAR [2, 5,

17]. However, the physiological and biochemical basis

for this phenomenon is not clear at present.
Pathological disorders caused by microbial agents
usually promote the development of hypersensitive
reactions within the infected plant tissues. If the pathogen is allowed to develop unchecked, necrotic lesions
develop, resulting in cell and tissue death [1]. It has
been demonstrated [10] that this sequence involves
destructive attack by free radicals mediated through
oxidative degradation of membrane lipids.
Similarly there is considerable evidence to suggest
that irreversible injury due to environmental stress is
caused by increased free radical titre and consequent
oxidation events which lead to degradation of biomolecules such as membrane lipids and proteins [15,
22, 23].
The similarity of the injury mechanism between
pathogenesis and stress leads us to hypothesise that
salicylic acid which induces resistance to disease also
confers tolerance to environmental stress. In this communication, evidence is provided that salicylic acid

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Table 1. Survival (%) of SA or ASA treated tomato and bean plants after heat, cold and drought stress
Heat
SI

Chilling
SD

SI

Drought
SD

SI

SD

Conc(mM)

Tomato

Bean

Tomato

Bean

Tomato

Bean

Tomato

Bean

Tomato

Bean

Tomato

Bean

SA

0
0.05
0.1
0.5
1.0

0
0
100
100
0

0
0
100
100
0

0
0
100
100
0

0
0
100
100
0

0
0
100
100
0

0
0
100
100
0

0
0
100
100
0

0
0
100
100
0

0
0
100
100
0

0
0
100
100
0

0
0
100
100
0

0
0
100
100
0

ASA

0
0.05
0.1
0.5
1.0

0
0
100
100
0

0
0
100
100
0

0
0
100
100
0

0
0
100
100
0

0
0
100
100
0

0
0
100
100
0

0
0
100
100
0

0
0
100
100
0

0
0
100
100
0

0
0
100
100
0

0
0
100
100
0

0
0
100
100
0

SI Seed Imbibed.
SD Soil Drenched.

(SA) and acetyl salicylic acid (ASA) provide multiple

stress tolerance in plants and that salicylic acid and
its derivatives regulate the expression of stress tolerance. This is the first report to suggest that SA or ASA
imbibed into a seed impart tolerance to a variety of
stresses in plants.

2. Materials and methods

To evaluate the hypothesis that SA and derivatives
can confer stress tolerance, experimentation was
conducted using bean (Phaseolus vulgaris L. cv.
brown beauty) and tomato (Lycopersicon esculentum
L. cv. romano) as test species. Plants were grown in
135 ml pots in a glasshouse maintained at ambient
temperature and humidity (in Perth, Western Australia). Fourteen day-old plants were soil-drenched
with 20 ml of distilled water or 0.05, 0.1, 0.5, 1.0
and 5.0 mM ASA or SA (dissolved in distilled water).
Alternatively, seeds were imbibed in the solutions or
in distilled water for 24 h and sown in pots.
One week after soil-drenching or three weeks after
the seed treatment, seedlings were subjected to heat,
cold and drought stresses. For heat treatment, seedlings were exposed to 54 0.5 C for 3 h with an
average light intensity of 40 Mol m2 sec1 and
then returned to room temperature. For chilling stress,
plants were exposed to 0 0.5 C in an incubator
with an average light intensity of 35 Mol m2 sec1
and 16/8 h light/dark photoperiod for two days. All
pots were saturated with water 2 h prior to and after

the imposition of heat and chilling stress treatments.

Drought stress was imposed by withholding water for
7 days, then on the 8th day all pots were watered until
saturation.
To evaluate the benefit of foliar spray applications plants were sprayed with either 0.5 mM ASA
or 0.5 mM SA, and subjected to stress treatments as
above. The nozzle of the hand held sprayer was adjusted to deliver 0.5 ml of solution per spray and each
plant received 1 ml of solution or distilled water for
control. One week after spraying, plants were exposed
to stress and scored for survival.
All treatments were assessed after 48 h and survival was determined by the ability of plants to regain
turgidity and resume normal growth after the stress
treatment. Damage was recorded as irreversible wilting, desiccation of leaves and presence of necrotic leaf
lesions. Randomized complete block design with 5
replicates was used in all experiments.

3. Results and discussion

Concentrations over 1 mM ASA or SA appeared to
have adverse effects and therefore lower concentrations were used in further experiments. There were no
observable differences in plant height, vigour or leaf
number between non treated and SA or ASA treated
plants (data not shown). Heat treatment at 54 C for
3 h induced severe wilting in controls of tomato and
bean plants and in plants treated with 0.05 mM SA
or ASA. The plants, seed imbibed or drenched with

159
0.1 mM or 0.5 mM SA or ASA did not exhibit signs of
wilting. After two days, plants which were not treated
with SA or ASA or which were treated with 0.05 mM
or 1.0 mM SA or ASA had not regained turgor in any
leaves and exhibited no signs of resprouting (Table 1).
The plants which were soil drenched with 0.5 mM SA
or ASA did not display any observable injury symptoms (Figure 1). Minor damage was visible in the
leaves of 0.1 mM treated plants, evident as desiccation
of leaf tips and some margins; however the injury was
transitory and the plants exhibiting these symptoms
recovered and were able to revive and resume growth.
Similar suppression of stress injury following
chilling treatment was observed in plants treated with
SA and ASA. Both tomato and bean plants subjected
to soil-drenching or seed treatment with 0.5 mM SA or
ASA did not display any injury symptoms after cold
stress treatment. Plants which received the 0.1 mM
SA or ASA treatments displayed necrotic lesions on
some leaves, but resumed growth after stress treatment (Table 1). The plants not treated with SA or
ASA, or treated with 0.05 mM or 1.0 mM SA or
ASA as soil-drench or seed treatment showed typical
symptoms of chilling injury with wilting of leaves,
necrosis and subsequent desiccation of most leaves
(Figure 1). One week after withholding water, the
plants not treated with SA or ASA as well as those
treated with 0.05 mM or 1.0 mM SA or ASA lost
turgor and wilted, whereas plants subjected to soildrench or seed treatment with 0.1 mM or 0.5 mM
SA or ASA retained a relatively high degree of turgidity. Most importantly, upon watering, 0.1 mM and
0.5 mM treated plants recovered completely within a
few hours, whereas untreated plants or those treated
with 0.05 mM or 1.0 mM SA or ASA retained wilted
leaves which subsequently desiccated (Table 1). Spray
treatment with 0.5 mM SA or ASA was also effective.
The concentration was selected based on the results
of previous experiments. All the SA or ASA treated
plants survived (100% survival) after heat, chilling or
drought stress while all the control plants died.
Enhancing stress tolerance in plants has major
implications in agriculture, horticulture, forestry as
well as in the re-establishment of natural vegetation.
However a simple method for inducing multiple stress
tolerance in plants without undesirable side effects has
not been available till now. Seed imbibition or drenching with triazole compounds such as paclobutrazol,
triademophone and uniconazole have been demonstrated to induce multiple stress tolerance in plants
[9, 12, 21]. Unfortunately, these compounds also

Figure 1. Bean plants A) exposed to heat stress B) pre-treated as a

soil drench with 0.5 mM ASA and exposed to heat stress C) exposed
to chilling D) pre-treated as a soil drench with 0.5 mM ASA and
subjected to chilling E) subjected to drought F) pre-treated as a soil
drench with 0.5 mM ASA and subjected to drought. Immediately
after the completion of heat, chilling or drought treatments all pots
were saturated with water (photographs were taken 48 h after the
completion of stress treatments).

160
inhibit gibberellin biosynthesis in plants and hence
have growth retarding effects. Salicylic acid and acetyl
salicylic acid are not known to retard plant growth and
no evidence of growth impairment of treated plants
over untreated controls was observed in this study.
The fact that seed imbibition of SA and ASA confers
tolerance to plants suggests that these molecules trigger the expression of the potential to tolerate stress
rather than having any direct effect as a protectant.
Some reports suggest that SA induces an oxidative
burst involving H2 O2 (hydrogen peroxide) accumulation which acts as the signal transducer for SAR
[3]. Neuenschwander et al. [18] did not observe major
changes in H2 O2 levels during the onset of SAR. The
exact mechanism of action of SA in inducing SAR
and its role in the transient increase of H2 O2 is still
cause for debate [4]. Prevention of oxidative damage
to cells during stress has been suggested as one of
the mechanisms of stress tolerance [12, 24] and this
level of protection is attributed to enhanced antioxidant activity [6, 21, 24]. Enhanced thermotolerance has
been observed in potato microplants grown on media
containing acetyl salicylic acid [14]. Increased thermotolerance in mustard seedlings sprayed with SA has
also been reported [7] and the tolerance was associated with changes in antioxidants such as glutathione
reductase, dehydroascorbate reductase and monodehydroascorbate reductase [6]. In pea seedlings SA
treatment decreased catalase and peroxidase levels
with concomitant increase in glutathione reductase
[26]. SA treatment also increased the level of reduced
glutathione (GSH) with an increase in the ratio of
reduced to oxidised glutathione (GSSG) indicating
higher antioxidant potential [26]. There are suggestions that salicylic acid acts as an antioxidant [4].
However the fact that seed imbibition with SA or ASA
provides stress tolerance in plants is more consistent
with a signalling role for the expression of tolerance
rather than a direct effect.
The other mechanisms of tolerance may involve
avoidance of lethal stress by altering cellular metabolism, synthesis of stabilising molecules to maintain
integrity and function of cellular membranes during
stress [27]. Stomatal resistance in SA treated mustard seedlings with enhanced thermotolerance was not
different from controls [7]. However detailed studies
on plant water relations after SA and ASA treatments are necessary to understand the events leading
to stress tolerance. Although the physiological and
biochemical basis for SA induced tolerance is not
clearly understood, we believe that a cascade of events

is triggered to provide multiple stress tolerance in

plants. Further research is warranted to elucidate the
physiological and biochemical mechanisms by which
SA induces tolerance to a variety of environmental
stresses. In addition research is required to elucidate
the reasons why plants treated with higher concentrations of SA or ASA (eg. 1.0 mM) were susceptible to
stress injury. However, the phenomenon that common
aspirin can be utilised to prevent crop losses during
stress may have significant practical application.

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