Environmental and Experimental Botany 66 (2009) 487–492

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Environmental and Experimental Botany

journal homepage: www.elsevier.com/locate/envexpbot

Physiochemical and antioxidant responses of the perennial xerophyte Capparis

ovata Desf. to drought
Ozden Ozkur, Filiz Ozdemir, Melike Bor, Ismail Turkan ∗
Department of Biology, Science Faculty, Ege University, Bornova 35100, Izmir, Turkey

a r t i c l e i n f o a b s t r a c t

Article history: Caper (Capparis ovata Desf.) is a perennial shrub (xerophyte) and drought resistant plant which is well
Received 7 November 2008 adapted to Mediterranean Ecosystem. In the present study we investigated the plant growth, relative
Received in revised form 31 March 2009 water content (RWC), chlorophyll fluorescence (FV /FM ), lipid peroxidation (TBA-reactive substances con-
Accepted 19 April 2009
tent) as parameters indicative of oxidative stress and antioxidant enzymes such as superoxide dismutase
(SOD), ascorbate peroxidase (APX), peroxidase (POX), catalase (CAT) and glutathione reductase (GR) in
Keywords:
relation to the tolerance to polyethylene glycol mediated drought stress in C. ovata seedlings. For induction
Caper
of drought stress, the 35 days seedlings were subjected to PEG 6000 of osmotic potential −0.81 MPa for
Drought stress
Antioxidative enzymes
14 days. Lipid peroxidation increased in PEG stressed seedlings as compared to non-stressed seedlings
Drought tolerance of C. ovata during the experimental period. With regard to vegetative growth, PEG treatment caused
decrease in shoot fresh and dry weights, RWC and FV /FM but decline was more prominent on day 14 of
PEG treatment. Total activity of antioxidative enzymes SOD, APX, POX, CAT and GR were investigated in C.
ovata seedlings under PEG mediated drought. Induced activities of SOD, CAT and POX enzymes were high
and the rate of increment was higher in stressed seedling. APX activity increased on both days of PEG
treatment, however, increase in GR activity was highest on day 14 of drought stress. We concluded that
increased drought tolerance of C. ovata is correlated with diminishing oxidative injury by functioning of
antioxidant system at higher rates under drought stress.
© 2009 Elsevier B.V. All rights reserved.

1. Introduction
Drought, in conjunction with coincident high temperature and
radiation, is considered as one of the most important environ-
mental extremes that constraints to plant survival and to crop
productivity in arid- and semi-arid regions (Chaves et al., 2003).
However, to cope with such combination of stresses, which are
known as drought stress, some plants such as Mediterranean xero-
phytes exhibit distinct resistant mechanisms which make them
good systems to understand physiological and biochemical mech-
anisms underlying drought tolerance of plants. These mechanisms
which are based upon osmotic adjustment, regulation of stomatal
opening, modification of cell wall characteristics and extensive root
system (Rhizopoulou and Psaras, 2003), all, involve in drought tol-
Abbreviations: RWC, relative water content; ROS, reactive oxygen species; SOD,
superoxide dismutase; CAT, catalase; APX, ascorbate peroxidase; GR, glutathione
reductase; POX, peroxidase; PEG, polyethylene glycol; BSA, bovine serum albumin;
NBT, nitroblue tetrazolium; EDTA, ethylenediamine-N,N,N ,N -tetraacetic acid; MDA,
malondialdehyde; DAB, diamino-benzidine tetra-hydrochloride; GSH, glutathione;
GSSG, oxidized glutathione; GDH, glutamate dehydrogenase; DW, dry weight; FW,
fresh weight; PAGE, polyacrylamide gel electrophoresis.
∗ Corresponding author. Tel.: +90 232 3884000x2443; fax: +90 232 3881036.
E-mail address: ismail.turkan@ege.edu.tr (I. Turkan).
erance of these plants, particularly during the summer when low
water availability is superimposed on high light and high temper-
atures at mid-day (Munne-Bosch and Penuelas, 2004).
Such as other environmental stressors, drought stress causes
also oxidative stress due to decreased stomatal conductivity which
restricts CO2 influx in to the leaves. Decreased leaf internal CO2
leads to the formation of reactive oxygen species (ROS) such as rad-
ical (O2 •− ), hydroxyl radical (OH), hydrogen peroxide (H2 O2 ) and
alkoxyl radical (RO) by enhanced leakage of electrons to molec-
ular oxygen. Chloroplasts, mitochondria and peroxisomes are the
major source of ROS in plant cells (Asada, 1999). Reactive oxygen
species have long been proposed as signal molecules that reg-
ulate various processes such as growth, development, responses
to biotic and abiotic environmental stimuli and programmed cell
death (Mittler et al., 2004; Apel and Hirt, 2004; Chung et al.,
2008). However, at high concentrations, these ROS can be toxic by
destroying normal metabolism through oxidative damage to lipids,
proteins and nucleic acids (Fridovich, 1986). Oxidative damage in
the plant tissue is alleviated by a concerted action of both enzymatic
and non-enzymatic antioxidant mechanism. These mechanisms
include ␤-carotens, alpha-tocopherol, ascorbate, glutathione and
enzymes including superoxide dismutase (SOD), peroxidase (POX),
ascorbate peroxidase (APX), catalase (CAT) and glutathione reduc-
tase (GR) (Smirnoff, 1993; Munne-Bosch and Penuelas, 2004). There

0098-8472/$ – see front matter © 2009 Elsevier B.V. All rights reserved.
doi:10.1016/j.envexpbot.2009.04.003
488 O. Ozkur et al. / Environmental and Experimental Botany 66 (2009) 487–492
are many reports in the literature that underline the intimate rela-
tionship between enhanced or constitutive antioxidant enzyme
activities and increased resistance to drought stress (Jagtap and
Bhargava, 1995; Türkan et al., 2005). Supporting this idea, enhanced
antioxidant defence under drought stress were also reported in
drought tolerant plants such as Mediterranean plants such as oak
(Quercus robur) (Schwanz and Polle, 2001), strawberry tree (Arbutus
unedo) and olive tree (Olea europeae) (Sofo et al., 2004).
Caper (Capparis ovata Desf.) which belongs to Cappareacea fam-
ily is a winter-deciduous perennial shrub (xerophyte) and drought
resistant plant, grows naturally and flower entirely during summer
in Mediterranean and semi-arid environments including Greece,
Cyprus and Turkey (Pascual et al., 2008). It can grow in places where
annual rainfall is about 350 mm and easily survive summertime
temperatures higher than 40 ◦ C (Barbera, 1991; Söyler and Arslan,
2000). Beside its use for soil erosion prevention, C. ovata is also
important economically since it can be used as drugs, cosmetics and
food (Matthaus and Özcan, 2005). Recently, it is widely cultivated
commercially in Morocco, Spain and Italy owing to the increasing
international demand for its pickled products (Pascual et al., 2008;
Ölmez et al., 2006). It also can resist against high salinity and grow
in poor-nutrient soils. Hence, to elucidate drought related traits of
this species is not only important to understand how drought tol-
erant plants are functioning in their natural environments but also
improve its agronomic characteristics. However, underlying charac-
teristics of physiological and biochemical response of C. ovata to the
drought is not known. Even, there are no data available related to
the relationship between drought stress and antioxidant defense in
this species. Therefore, in present study, the potential role of antiox-
idant enzymes in enhancing its protecting C. ovata from oxidative
stress of drought was examined by analyzing enzyme activities such
as SOD, CAT, POX, APX and GR. We also investigated the effects of
drought which is created by application of PEG on the basic param-
eters such as growth, relative water content (RWC) and chlorophyll
fluorescence (FV /FM ), in leaves of C. ovata.
2. Materials and methods
2.1. Plant material and treatments
C. ovata seedlings which were obtained from Aegean Agricul-
tural Research Institute-Menemen, İzmir (AARI) were used in this
study. Seedlings were sown into the pots (20 cm × 30 cm) filled with
perlite and grown under controlled conditions (light/dark regime
of 16/8 h at 27/22 ◦ C, relative humidity of 60–70%, photosynthetic
photon flux density of (PAR) 350 ␮mol m−2 s−1 ). After germina-
tion, seedlings were watered regularly with half-strength Hoagland
solution. Polyethylene glycol (PEG) 6000 treatment started when
plants were 35 days old by adding 20% PEG which is equivalent to
an osmotic potential of −0.81 MPa, into the half-strength Hoagland
solution. Seedlings were watered regularly with the half-strength
Hoagland solution containing either 0 or 20% PEG in 2-day intervals
extended for 14 days.
2.2. Growth parameters
Twenty seedlings from each treatment were sampled randomly
at the 0, 7 and 14th days. Fresh weights (g) of shoots were recorded
and for dry weight (g) determination samples were oven dried at
70 ◦ C for 72 h and then weighed.
2.3. Relative water content (RWC)
Leaf samples which were collected at the 0, 7 and 14th days
of PEG-treatment were used for RWC assay. After fresh weight
determination, they were floated on deionised water for 5 h under
low irradiance. The turgid leaf was quickly blotted dried prior to
the determination of turgid weight. Dry weights of leaves were
determined after oven-drying at 70 ◦ C for 72 h. RWC was calcu-
lated according to Smart and Bingham (1974), using the following
formula:
 fresh weight − dry weight 
RWC (%) = × 100
turgid weight − dry weight
2.4. Chlorophyll fluorescence
Leaf of seedlings from six individual plants per treatment was
used for chlorophyll fluorescence analysis. Prior to fluorescence
measurements, a 1 cm2 circular surface of the upper face of excised
leaf was dark adapted for 30 min using dark leaf clip. The basal
non-variable chlorophyll fluorescence level (F0 ), the maximal fluo-
rescence induction (FM ), variable fluorescence (FV ) and the ratio of
FV /FM were determined by a Plant Efficiency Analyser (HANSATECH
Inst. Ltd., Norfolk, UK). More especially, photochemical efficiency of
PS-II (maximum quantum yield FV /FM ) was figured.
2.5. Lipid peroxidation
Lipid peroxidation was determined by estimating the TBA-
reactive substances (TBARS) content in 0.5 g leaf fresh weight
according to Madhava Rao and Sresty (2000). TBARS are products
of lipid peroxidation by thiobarbituric acid reaction. The concentra-
tion of TBARS (nmol g FW−1 ) was calculated from the absorbance
at 532 nm (correction was done by subtracting the absorbance at
600 nm for unspecific turbidity) by using extinction coefficient of
155 mM−1 cm−1 .
2.6. Enzyme assays
Fresh leaf samples of both species obtained at 0, 7 and 14 days
after PEG treatment were used for enzyme analysis. Leaves were
frozen in liquid nitrogen immediately after harvesting and stored at
−20 ◦ C until enzyme assays. 1 g leaves homogenized in 3 mL 0.05 M
Na phosphate buffer (pH 7.8) including 1 mM EDTA and 2% (w/v)
PVPP. The homogenate were centrifuged at 13,000 × g for 40 min at
4 ◦ C. Supernatant was used for enzyme activity and protein content
assays. All assays were done at 4 ◦ C. Total soluble protein contents of
the enzyme extracts were determined according to Bradford (1976)
using BSA as a standard. All spectrophotometric analyses were con-
ducted on a Shimadzu (UV-1600) spectrophotometer.
Superoxide dismutase (SOD; EC 1.15.1.1) activity assay was based
on the method of Beauchamp and Fridovich (1971) which mea-
sures the inhibition in the photochemical reduction of nitroblue
tetrazolium (NBT) spectrophotometrically at 560 nm. One unit of
enzyme activity was defined as the quantity of SOD required to pro-
duce a 50% inhibition of reduction of NBT and the specific enzyme
activity was expressed as units mg−1 protein. The reaction mix-
ture contained 50 mM Na phosphate buffer (pH 7.8), 33 ␮M NBT,
10 mM l-methionine, 0.66 mM EDTA and 0.0033 mM riboflavin.
Reactions were carried out at 25 ◦ C, under light intensity of about
300 ␮mol−1 m−1 s−1 through 10 min.
Catalase activity (CAT EC 1.11.1.6) was done according to
Bergmeyer (1970) which measures the decline of the extinction of
H2 O2 at the maximum absorption at 240 nm. The reaction mixture
contained 0.05 M Na phosphate buffer (pH 7.0) with 1 mM EDTA and
H2 O2 (3%). The decrease in the absorption was followed for 3 min
and ␮mol H2 O2 destroyed per min was defined as one unit of CAT.
Ascorbate peroxidase (APX; EC 1.11.1.11) activity was done
according to Nakano and Asada (1981). The assay depends on
the decrease in absorbance at 290 nm as ascorbate was oxidized
(extinction coefficient of 2.8 mM−1 cm−1 ). The reaction mixture
 O. Ozkur et al. / Environmental and Experimental Botany 66 (2009) 487–492
contained 50 mM Na–phosphate buffer (pH 7.0), 0.5 mM ascorbate,
0.1 mM EDTA Na2 and 1.2 mM H2 O2 . One enzyme unit was defined
as ␮mol mL−1 oxidized ascorbate per min.
Glutathione reductase (GR; EC 1.6.4.2) activity was measured
according to Foyer and Halliwell (1976) which depends on the
rate of decrease in the absorbance of oxidized glutathione (GSSG)
at 340 nm. The reaction mixture contained 25 mM Na–phosphate
buffer (pH 7.8), 5 mM GSSG, 1.2 mM NADPHNa4 . The reaction was
carried out for 3 min and activity of GR was calculated from the
reduced GSSG concentration by using the extinction coefficient
6.2 mM−1 cm−1 . One enzyme unit was defined as ␮mol mL−1 oxi-
dized GSSG per min.
Peroxidase activity (POX; EC 1.11.1.7) was based upon the method
as described by Herzog and Fahimi (1973) which measures the
increase in absorbance at 465 nm, by the rate of formation of 0.15 M
Na–phosphate citrate buffer the oxidized DAB. The reaction mixture
contained DAB solution (dissolved gelatine solution and contained
50%, w/v) and 0.6% H2 O2 . The increase in A465 was followed for
3 min. One enzyme unit was defined as ␮mol mL−1 destroyed H2 O2
per min.
The specific enzyme activity for all enzymes was expressed as
in unit mg−1 protein.
2.7. Statistical analysis
All analyses were done on a completely randomized design.
All data obtained was subjected to one-way analysis of variance
(ANOVA) and the mean differences were compared by lowest stan-
dard deviations (LSD) test. Each data point was the mean of six
replicates (n = 6), except for dry and fresh weights of C. ovata
seedlings (n = 12). Comparisons with P values <0.05 were consid-
ered significantly different. In all the figures the spread of values is
shown as error bars representing standard errors of the means.
3. Results
The level of TBA-reactive substances (TBARS) is given in Fig. 1.
In C. ovata, the levels of TBARS showed variation with age and also
PEG treatment. In the control leaves of C. ovata, a small ‘age depen-
dent increase’ in TBARS levels became apparent after 14 days of
PEG treatment. Where as, TBARS levels significantly increased in
the leaves of drought-stressed C. ovata on day 7 and 14 as compared
to TBARS levels in control groups.
Shoot growth of C. ovata was followed by measuring fresh weight
(FW) and dry weight (DW) on days 0, 7 and14 (Table 1). Drought cre-
Fig. 1. The effect of PEG treatment on lipid peroxidation (MDA content) in leaves
of Capparis ovata. Data represents the average of two experiments with three repli-
cates. Vertical bars indicate ±SE. Values sharing a common letter are not significantly
different at p < 0.05.
489
Table 1
The effect of PEG treatment during experimental period (0, 7 and 14th days) on
growth parameters shoot fresh and dry weights (g), relative water content (RWC)
and photosynthetic efficiency (FV /FM ) on Capparis ovata. Means ± SD based on twelve
replicates (n = 12) for fresh weight and dry weight, six replicates (n = 6) for RWC and
photosynthetic efficiency (FV /FM ) are presented. Values sharing a common letter are
not significantly different at p < 0.05.
RWC (%) FV /FM Fresh weight (g) Dry weight (g)
Control 0 83.13 ± 2.78b 0.86 ± 0.011ab 0.0872 ± 0.015a 0.0182 ± 0.003b
Control 7 83.71 ± 2.22b 0.86 ± 0.011ab 0.1140 ± 0.005b 0.0202 ± 0.004c
PEG 7 75.96 ± 2.44a 0.84 ± 0.006a 0.0898 ± 0.009a 0.0145 ± 0.010a
Control 14 84.02 ± 1.14b 0.85 ± 0.008ab 0.1487 ± 0.012c 0.0220 ± 0.002d
PEG 14 76.29 ± 2.30a 0.83 ± 0.042a 0.1148 ± 0.004b 0.0146 ± 0.003a
ated by application of 20% PEG 6000 significantly decreased fresh
and dry weights of C. ovata seedlings. Under drought stress, FW
decreased by about 21 and 23%, whereas the decrease in DW were
about by 28 and 34% on day 7 and 14 of PEG treatments, respectively,
as compared to their controls.
The RWC, as indicated by the extent of dehydration were used
to assess cellular damage. Over the experimental period (14 days),
RWC in the leaves of C. ovata declined under drought stress (Table 1).
However, the results related to the RWC showed similar trend, with
about 8.0% decline on both days 7 and 14 by PEG treatments.
Under drought conditions, a significant alteration in FV /FM was
found in the leaves of C. ovata (Table 1). As compared to control,
FV /FM decreased by 2.90 and 2.59% on day 7 and 14 of PEG treatment,
respectively.
In the present study we also observed that drought resulted
in higher enzyme activities of SOD, POX, CAT, APX and GR in the
leaves of C. ovata. Total SOD activities increased in response to PEG
treatments (Fig. 2). The increases in SOD activity were 1.7-fold and
1.8-fold in the leaves of C. ovata on days 7 and 14 of the drought
application as compared to their controls. APX activity showed no
differences in control plants during the experimental period (14
days). However, high osmoticum (20% PEG) caused 5 and 4 folds
increase in APX activity compared to that of the control’s on days 7
and 14, respectively (Fig. 2). The increased APX activity observed in
this study might have been due to the increased H2 O2 production
under drought stress. POX activity increased significantly in C. ovata
both on day 7 and 14 under osmotic stress (Fig. 2). The increase has
been 2 and 3 folds on day 7 and 14 of the treatment as compared
to controls. PEG treatment result in a significant increase in total
CAT activity in the leaves of C. ovata (Fig. 2). The CAT activity raised
1.5 fold on both day 7 and 14 of PEG treatments compared to their
controls. GR was also significantly increased in drought-stressed
seedlings of C. ovata on day 14 and the rate of increment was 3-fold
(Fig. 2).
4. Discussion
Drought poses the most important environmental constraint
to plant survival and crop productivity in natural and agricul-
tural habitats (Chaves et al., 2003). Due to its osmotic effect it can
induce a wide number of responses ranging from growth inhibi-
tion and synthesis of some non-toxic compounds to increase the
osmotic potential of the cell and thus allow metabolic processes
to continue to enhancement of some antioxidant enzyme activities
(Türkan et al., 2005). In present study, drought adversely effected
seedling growth of C. ovata as it was evident from decreased dry and
fresh weights of the leaves. Inhibition in growth of drought toler-
ant Mediterranean shrubs A. unedo L. (Munne-Bosch and Penuelas,
2004) and Atriplex halimus L. (Hassine et al., 2008) were also
observed in previous studies. Although C. ovata leaves show some
degree of dessication, it was able to maintain its internal water sta-
tus at very similar degree at extended period of drought stress. In
490 O. Ozkur et al. / Environmental and Experimental Botany 66 (2009) 487–492

Fig. 2. The effect of PEG treatment on SOD, APX, POX, CAT and GR activities in leaves of C. ovata on days 0, 7 and 14. Data represents the average of two experiments with
three replicates. Vertical bars indicate ±SE and values sharing a common letter are not significantly different at p < 0.05.

accordance with this result, Ana Lúcia et al. (2002) also observed
a decline in leaf water potential even in drought tolerant clone
of Coffea canephora under water deficit. A decrease in RWC levels
were also observed in A. unedo which can withstand drought condi-
tions of Mediterranean environments (Munne-Bosch and Penuelas,
2004).
Decrease in stomatal conductance leading to exposure of excess
energy at chloroplasts and over reduction of reaction centers in
PSII limits photosynthesis in plants under drought stress (Demmig-
Adams and Adams, 1992). Photosynthetic efficiency which was
measured as FV /FM values, decreased under drought stress on both
days as compared to that of non-stressed plants in the leaves
of C. ovata. Sofo et al. (2007) also found decreased FV /FM values
under severely stressed olive plants. Photosynthetic efficiency also
reduced in the seedlings of 20% PEG treated A. halimus plants
(Hassine et al., 2008) and drought-stressed Salvia officinalis (Abreu
and Munné-Bosch, 2008). A decrease in photochemical efficiency
(FV /FM ) is regularly described in temperature and drought stress
responses and may be interpreted as a photo-acclimation process,
more than a symptom of damage since it was also the case in other
Mediterranean species such as Phyllirea latifolia, A. unedo, S. offici-
nalis and Cistus salvifolius exposed extreme heat-wave of Summer
2003 in Southwestern Europe (Munne-Bosch and Penuelas, 2004;
Abreu and Munné-Bosch, 2008; García-Plazaola et al., 2008).
Increment in TBA-reactive substances (TBARS) is a good reflec-
tion of oxidative damage to membrane lipids and other vital
 O. Ozkur et al. / Environmental and Experimental Botany 66 (2009) 487–492
molecules such as proteins, DNA and RNA. In our study, TBARS levels
increased in the leaves of C. ovata under drought stress which is in
agreement with results of other studies (Schwanz and Polle, 2001;
Ana Lúcia et al., 2002; Sofo et al., 2004). Peroxidation of membrane
lipids may result in enhanced membrane fluidity, which may led
to enhanced electrolyte leakage, as suggested by Thompson et al.
(1987) and support the hypothesis that drought stress can induce
membrane lipid peroxidation.
Under drought conditions, in the electron transport chain, elec-
trons that have no access for their final destination, CO2 are directed
to the reduction of molecular oxygen to from superoxide and
hydrogen peroxide (Miyake and Yokota, 2000). Plants have an
efficient system for decomposing reactive oxygen species, using
the enzymes superoxide dismutase (SOD) and ascorbate peroxi-
dase (APX) in chloroplasts (Asada, 1999). In present study, drought
resulted in higher enzyme activities of SOD, POX, CAT, APX and
GR in the leaves of C. ovata seems to be the physiological adap-
tive mechanisms to regulate its redox status under drought stress
conditions.
Among antioxidant enzymes mentioned above, SOD converts
the toxic O2 − to H2 O2 which must be scavenged to the O2 and
H2 O by the antioxidant enzymes such as CAT, POX and APX. SOD in
higher plants exists in multiple isoforms that are developmentally
regulated and highly reactive to exogenous stimuli (Pan et al., 2006).
Induction of total SOD activity which was correlated with increased
protection from damage associated with oxidative stress implies
that enhancement of SOD scavenge O2 •− radicals to protect C. ovata
from oxidative damage. Similar to our findings Schwanz and Polle
(2001) found significant increase in SOD levels in water stressed,
drought tolerant Mediterranean shrub Q. robur. SOD activity was
also increased by drought stress in shoots of other Mediterranean
plants such as Myrtus communis and Phillyrea angustifolia (Caravaca
et al., 2005).
APX which is primarily located both in chloroplasts and cytosol
acts as a key enzyme of the glutathione–ascorbate pathway. It scav-
enges peroxides by converting ascorbic acid to dehydroascorbate
(Navari-Izzo et al., 1997); it is one of the most important enzymes
playing a vital role in eliminating toxic H2 O2 from plant cell in
Asada–Halliwell pathway (Foyer et al., 1994). In the present study,
drought stress increased total APX activity differed prominently in
stressed leaves as compared to that of non-stressed leaves. Higher
activity of APX in stressed leaves suggests a more effective H2 O2
removal which might be produced by an enhanced activity of SOD
in C. ovata under drought conditions. A higher APX activity was
also reported in tobacco BY-2 cell cultures (Bueno et al., 1998) and
tepary bean under drought stress (Türkan et al., 2005). Parallel to
our results, an upregulation in APX activity was also observed in
drought-stressed olive (Sofo et al., 2007) and maize (Pastori and
Trippi, 1992).
POX is among the major enzymes that scavenge H2 O2 in chloro-
plasts which are produced through dismutation of O2 − catalyzed
by SOD (Asada and Takahashi, 1987). In tolerant plant species, POX
activity was found to be higher, providing protection against the
oxidative stress (Scalet et al., 1995). Remarkably higher levels of POX
in drought-stressed seedlings of C. ovata might also be considered
as a higher capacity to decompose H2 O2 more rapidly. Supporting
our results, Yang et al. (2008) also reported induced higher activi-
ties in Picea aspertata seedlings under drought stress supplemented
with a high light. In one of our previous studies we also observed
that drought resistant tepary bean showed a higher constitutive and
induced activity of POX under drought stress (Türkan et al., 2005).
CAT eliminates H2 O2 by breaking it down directly to water and
oxygen (Tsang et al., 1991). In our study, total CAT activity was
enhanced by drought in the leaves of C. ovata. These results are
consistent with the reports from Mittler and Zilinkas (1993) and
Sofo et al. (2007) who found that with drought stress, CAT activities
491
increased in drought-stressed peas and olive, respectively. When
one considers that CAT acts in peroxisomes, it is logical to suggest
that photorespiration in the leaves of C. ovata was also affected by
PEG treatment.
In the present study, drought stress result in a significant
increase in GR the activity which is of another major enzyme that
has a role in the H2 O2 scavenging in Asada–Halliwell pathway
in plant cells (Noctor et al., 2002). This might be due to main-
tain a high ratio of NADP+ /NADPH, therefore ensuing availability
of NADP+ to accept electrons from photosynthetic electron trans-
port chain and to facilitate the regeneration of oxidized ascorbate
(Noctor et al., 2002; Yang et al., 2008). We could conclude that
ascorbate–glutathione cycle efficiently eliminates the deleterious
effects of reactive oxygen species in the leaves of C. ovata under
drought stress. Drought induced increase in GR was also observed
in Picea asperata seedlings under highlight conditions. Parallel to
our results GR activity significantly increased in drought tolerant
maize strain under water scarcity (Pastori and Trippi, 1992).
5. Conclusion
In conclusion, as a whole, the antioxidant system of C. ovata func-
tioned at higher rates to restrain an increased ROS formation under
drought. This seemed to be evident by evaluating the extent of cellu-
lar damages, which was less remarkable under stress. These results
are in good agreement with several studies indicating that drought
generally induces antioxidative system in Mediterranean plants and
increased drought tolerance is correlated with diminishing oxida-
tive injury (Pastori and Trippi, 1992; Smirnoff, 1993; Polle, 1997;
Schwanz and Polle, 2001).
Acknowledgment
This work was supported by grant-in-aids 2008-FEN-039 from
Ege University Research Foundation.
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