See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/258339742

Effect of heavy metals on plasma membrane lipids and antioxidant enzymes

of Zygophyllum

Article · January 2012

CITATIONS READS

5 353

1 author:

M. M. F. Mansour
Ain Shams University
55 PUBLICATIONS   1,460 CITATIONS   

SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Compatible solutes in saline conditions: an assessment of the current state of knowledge View project

All content following this page was uploaded by M. M. F. Mansour on 02 June 2014.

The user has requested enhancement of the downloaded file.

 EurAsian Journal of BioSciences
Eurasia J Biosci 6, 1-10 (2012)
DOI:10.5053/ejobios.2012.6.0.1

Effect of heavy metals on plasma membrane lipids

and antioxidant enzymes of Zygophyllum species
Amal Ahmed Morsy1, Karima Hamid Ali Salama2, Hend Ahmed Kamel1,
Mohamed Magdy Fahim Mansour2*
1Department of Botany, Faculty of Science, Ain Shams University, Cairo, Egypt
2Department of Biology, Faculty of Science, Taif University, Taif, Saudi Arabia

*Corresponding author: mf_mansour@yahoo.com

Abstract
Background: Heavy metals are major environmental pollutant when they present in high
concentration in soil and have toxic effects on growth and development of plants. Industrial
activities result in heavy metal pollution of large areas of land, which greatly affects natural
vegetation. Understanding the mechanism of how plants combat heavy metals adverse effects is
hence of great importance.
Materials and Methods: Two different localities were chosen; one locality was in the vicinity of
gypsum factory and the other one was 25 km away from the factory. Two Zygophyllum species (Z.
album and Z. coccineum) were naturally grown in the studied areas. The effects of soil heavy metal
stress on shoot heavy metal concentrations, lipid peroxidation, antioxidant enzyme activities and
the root plasma membrane (PM) lipid composition were analyzed.
Results: Heavy metal concentrations and Lipid peroxidation increased in the shoot of both species
grown in the polluted area. The activities of ascorbate oxidase (ASO), guaiacal peroxidase (GPX),
ascorbate peroxidase (APX) and superoxide dismutase (SOD) were increased whereas these of
catalase (CAT) were decreased in both species under the polluted conditions. PM total lipids,
phospholipids, glycolipids and sterols were decreased in Z. album and Z. coccineum as a result of the
polluted soil. Heavy metal stress increased phosphatidylethanolamine (PE) and decreased
phosphatidylinositol (PI) and phophatidylglycerol (PG), with no significant change in
phosphatidylcholine (PC) in the root PM of both species. Phosphatidylserine (PS) decreased in the
PM of Z. album whereas it increased in the PM of Z. coccineum under the pollution conditions. Heavy
metal stress changed the composition and concentration of fatty acids of the root PM, resulting in
increased sat/unsat ratio of both species.
Conclusion: the results suggest that efficient antioxidant machinery and favorable PM lipid
homeostasis are important to enable Zygophyllum species to withstand the prevailing heavy metal
stress.
Keywords: Antioxidant enzyme, fatty acid, heavy metal, lipid peroxidation, plasma membrane lipid,
Zygophyllum sp.

Morsy AA, Salama HHA, Kamel HA, Mansour MMF (2012) Effect of heavy metals on plasma
membrane lipids and antioxidant enzymes of Zygophyllum species. Eurasia J Biosci 6: 1-10.

DOI:10.5053/ejobios.2012.6.0.1 ©EurAsian Journal of BioSciences

agricultural practices and industry (Khan et al. 2000,

INTRODUCTION
Heavy metals are defined as group of elements
that have specific weights higher than about 5
g/cm3. Iron, Mn, Mo, Ni, Zn and Cu are essential
micronutrients that required for normal growth and
metabolic processes in plants (Vangronsveld and
Clijsters 1994). Cd, Pb, Cr and Hg are nonessential
and highly toxic for plants (Devi and Prasad 1998,
Sebastiani et al. 2004, Rai et al. 2004). Heavy metals
inhibit physiological processes such as respiration,
photosynthesis, cell elongation and affect plant
water relationship as well as mineral nutrition
(Vangronsveld and Clijsters 1994, Zornoza et al.
2002). Large areas of land are contaminated with
heavy metals resulting from urban activities,
Ciemense 2001).
Damage of the cellular membranes, especially for
the plasma membrane, is one of the primary events
in heavy metal toxic effects in plants (Harwood
1995, Janicka et al. 2008). Heavy metals disrupt
cellular membranes resulting in the conversion of
unsaturated fatty acids into small hydrocarbon
fragments such as malondialdehyde (Tappel 1973,
Kappus 1985). Metal ions can also replace calcium
ions at its essential sites on the membranes (Breckle
Received: September 2011
Accepted: January 2012
Printed: January 2012

1
EurAsian Journal of BioSciences 6: 1-10 (2012)
and Kahle 1991). Furthermore, plant plasma
membrane is dynamic and its lipid composition/
structure is changed with variations in the external
environment. Plasma membrane changes might
have an adaptive value or injuriously affects
membrane properties and functions (Mansour et al.
2003, Mansour and Salama 2004).
Reactive oxygen species (ROS) are regarded as
the main source of damage to cells under the biotic
and abiotic stresses (Mittler 2002, Candan and
Tarhan 2003, Gara et al. 2003, Vaidyanathan et al.
2003). This is because under normal conditions
production and destruction of ROS is well regulated
in cell metabolism (Mittler 2002). When a plant is in
the harsh conditions, ROS production will overcome
scavenging systems and oxidative stress will burst.
ROS includes superoxide (O2.-), hydrogen peroxide
(H2O2) and hydroxyl radical (HO·) (Mittler 2002). ROS
are highly cytotoxic and can seriously react with vital
bio-molecules such as lipids, proteins, nucleic acid,
these can cause lipid peroxidation, protein
denaturation and DNA mutation (Mittler 2002,
Quiles and López 2004). Plants have developed
various protective mechanisms to eliminate or
reduce ROS deleterious effects (Mittler 2002, Beak
and Skinner 2003). Antioxidant system consists of
enzymatic and non-enzymatic compounds.
Enzymatic antioxidants include superoxide
dismutase, catalase, peroxidases, glutathione
reductase and ascorbate reductase (Mittler 2002,
Candan and Tarhan 2003). Non-enzymatic
antioxidants are ascorbate, glutathione, tocopherol
and carotenoids.
The current study was undertaken to analyze the
changes in the heavy metal composition, plasma
membrane lipids, lipid peroxidation and antioxidant
enzymes of two Zygopyllum species (Zygophyllum
album and Zygophyllum coccinum) grown in their
natural habitats in absence and presence of heavy
metal pollution. These changes will be correlated
with the responses of the two Zygopyllum species
grown in polluted and non-polluted habitats.
2
Morsy et al.
MATERIALS AND METHODS
The study areas and plant materials
The present work was conducted in two different
desert habitats at south Sinai, Egypt; the first habitat
was Ras Malaab where the factory of gypsum exists.
The second habitat was Wadi Thal placed 25 km
away from the factory. The plant species used in the
present investigation were Zygophyllum album L.
and Zygophyllum coccineum L.
Soil samples
The samples of two composite soil were collected
at three depths (0-20 cm, 20-40 cm and 40-60 cm).
One sample was collected near an industrial effluent
channel and the other sample was collected 25 km
away. The soil samples collection were replicated
three times over space. Granulometric analyses of
the soil samples were accomplished using the sieve
method according to Jackson (1967). Soil-water
extract (1:10) was used for the different
determinations as described by Richards (1954). Al,
Cu, Zn, and Fe were determined using atomic
absorptions spectrophotometer (Unicam 929 AA,
UK) as described by Black (1965).
Plant samples
The samples of shoot and root of both species
were collected from their natural vegetation in Ras
Malaab and Wadi Thal. The samples were replicated
three times over space. The samples were kept in
liquid nitrogen immediately after collection, and
then stored under -80°C for the further analysis.
Determination of antioxidant enzyme
activities
The shoots were homogenized in a chilled mortar
for 30s using 50 mM potassium phosphate buffer
(pH 7). Homogenates were centrifuged at 15,000 g
at 4°C and the supernatant was used for enzyme
activities. The activity of APX was measured
according to Zhang et al. (2007). The reaction
mixture contained 50 mM potassium-phosphate
buffer, 0.5 mM L-ascorbate, 0.1 mM H2O2 and the
enzyme extract. H2O2 dependent oxidation of
ascorbate was followed by a decrease in absorbance
at 290 nm. APX activity was expressed as the
absorbance decrease (ΔE) min-1 g-1 F. W. GPX activity
was estimated according to Hammerschmidt et al.
EurAsian Journal of BioSciences 6: 1-10 (2012)
(1982). The reaction mixture contained 25 mM
potassium phosphate buffer (pH 7.0), 0.2 mM
guaiacol, 0.09 mM H2O2 and the enzyme extract.
H2O2-dependent oxidation of guaiacol was followed
by an increase of absorbance at 470 nm. Enzyme
activity was calculated as the increase of absorbance
(ΔE) min-1 g-1 F. W. For ASO, the method of Chinoy et
al. (1976) was used. The reaction mixture consisted
of 1 mL of 0.1 mM ascorbic acid, 1 mL of 0.5 mM
phosphate buffer (pH 6.8) and 2 mL of enzyme
extract. This reaction mixture was incubated at 37°C
for 25 min. After incubation, the 2, 6-dichlorophenol
indophenol was added and the absorbance was
measured at 620 nm. CAT activity was assayed
spectrophotometrically (Hitachi U-3000, Japan) by
measuring the decrease in absorbance at 240 nm
because of H2O2 decomposition (Zhang et al. 2007).
SOD activity was determined by measuring the
inhibition of the auto-oxidation of pyrogallol using a
method described by Zhang et al. (2007). Ten mL of
the reaction mixture comprised: 3.6 mL of distilled
water, 0.1 mL of the enzyme extract, 5.5 mL of 50
mM phosphate buffer (pH 7.8) and 0.8 mL of 3 mM
pyrogallol (dissolved in 10 mM HCl). The rate of
pyrogallol reduction was measured at 325 nm using
UV- spectrophotometer. One unit of enzyme activity
was defined as the amount of the enzyme that
resulted in 50% inhibition of the auto-oxidation rate
of pyrogallol at 25°C (Zhang et al. 2007). The enzyme
activity was expressed as unit/mg protein.
Determination of malondialdehyde (MDA)
MDA content, as an indicator of lipid
peroxidation, was determined using the procedure
described by Zhang et al. (2007). The plant shoot was
homogenized in 1% trichloroacetic acid and then
centrifuged at 10,000 rpm for 15 min. Supernatant
was heated with 0.05 thiobarbituric acid for 30 min
at 95°C. The heated supernatant was then
centrifuged at 5000 rpm for 5 min and the
absorbance was measured at 532 and 600 nm.
Plasma membrane isolation
Two-phase partitioning method (Mansour et al.
2002, Salama et al. 2007) was used to isolate the
root PM of Zygophyllum album and Zygophyllum
coccinum. The root was washed in cold bidistilled
water and homogenized in a blender with a
Morsy et al.
homogenization medium (the pH was adjusted to 7.5
with Hepps-KOH): 250 mM sucrose, 5 mM EGTA, 5
mM EDTA, 10 mM KF, 25 mM MOPS, and 1 mM PMSF.
Two mM of DTT was added as powder. The
homogenization of the root tissue was carried out
three times for 20-s each with a 10-s pause. The
homogenate was filtered through miracloth. A new
homogenization medium was added to the tissue
and was homogenized once again. The pooled
solution was centrifuged at 10,000 g for 20 min. The
supernatant containing the PM and other
membranes was then centrifuge at 50,000 g for 1 h.
The microsomal membrane pellet was resuspended
in 5 mM potassium phosphate buffer (pH 7.8)
containing 250 mM sucrose and 4 mM KCl. The PM
were prepared by partitioning of microsomal
suspension in 27 g aqueous polymer two-phase
system containing 6.5% dextrane T-500 (Pharmacia),
6.5% polyethylene glycol 3350 (Sigma) in 250 mM
sucrose, 5 mM potassium phosphate buffer (pH 7.8),
4 mM KCl and 25 mM DTT. Into the two-phase
system 108 μL of the solution (333 mM DTE and 33.3
mM EDTA) was added, mixed thoroughly and
centrifuged at 1,500 g for 5 min. The microsomal
pellet was subjected to three successive phase
partitioning steps. The upper phase, containing the
PM fraction, centrifuged at 50,000 g for 1 h and the
pellet resuspended in 1 mL of storage buffer (pH
7.5). All steps of the PM isolation were carried out at
0-4°C. The purity of our PM preparation was based
on Mansour et al. (1994).
PM lipid extraction and separation
Boiled isopropanol was immediately added to the
PM suspension to inhibit the activity of lipase (Kates
1972). Lipids were then extracted with 3.75 mM
chloroform: isopropanol (2:1, v/v) and 2.25 mL of 0.1
M KCl was added to enhance the chloroform phase
separation. Then, the mixture was centrifuged in
cold room at 1,000 g for 5 min. The upper water
phase was re-extracted with 2 mL chloroform. The
first and second chloroform phases (containing
lipids) were collected and dried under CO2 stream.
The dried lipids were dissolved in 2.5 mL chloroform
and stored at -80°C until analysis.
Determination of PM fatty acids
The method of Mansour et al. (2002) was used for
3
EurAsian Journal of BioSciences 6: 1-10 (2012)
the analysis of PM fatty acids. One mL of lipid
extract, 6 mL of benzene and 1.5 mL of 10% alcoholic
KOH were mixed together. The tubes were refluxed
for 4 h in a boiling water bath and then the mixture
was evaporated. Excess of diethyl ether was added
and shaken well. The organic phase (upper phase)
was pipetted and the aqueous phase (lower phase)
was further washed three times with diethyl ether.
The aqueous phase containing fatty acids was
acidified by 1 N H2SO4 and the pH was adjusted to
two. The sample was methylated by the addition of
2.5 mL of H2SO4 and 50 mL methanol and refluxed
for 24 h. The mixture was evaporated; excess of
diethyl ether was added and shaken well. The
organic phase was evaporated to determine the
different fatty acids. The fatty acid methyl esters
were determined by gas chromatography (HP-5890,
Little Falls, DE) equipped with a flame-ionization
detector. An HP-FFAP (free fatty acid phase) column
(25 m, 0.3 mm diameter) was used with helium (35
cm s-1) as a carrier gas. The injector temperature was
250°C, detector temperature was 260°C, and the
temperature program during the analysis went from
50 to 240°C (7°C min-1), after which temperature was
kept constant at 240°C for 30 min.
Determination of PM phospholipids
Phospholipids (polar lipids) were assayed
according to Deinstrop and Weinheim (2000). The
lipid extract was spotted along a glass thin layer
chromatography plate (TLC, Merk, Germany).
Phospholipid classes were separated by two-
dimensional TLC with solvent mixtures of
chloroform: methanol: deionized bidistilled water
(75: 25: 2.5, v/v) in the first direction. After allowing
sufficient time for drying, the plate was developed
at right angles to the first development in
chloroform: methanol: acetic acid: water (80: 9: 12: 2,
v/v) for the second direction. After separation of
phospholipids, spots were located by exposure to I2
vapor. Individual phospholipids were identified by
co-chromatography with authentic standards. The
area on TLC corresponding to each individual
phospholipids was marked and scraped into a test
tube and 0.5 mL HCl was added to neutralize silicate.
All tubes were heated for 10 min in a sand bath.
After cooling, 0.26 mL of 70% perchloric acid was
4
Morsy et al.
added. The tubes were closed with a glass covers
and heated at 180°C for 20 min and cooled. Next, 3.3
mL of deionized bi-distilled water, 0.4 mL of 10%
ascorbic acid were added. After each addition, the
contents of the tube were mixed thoroughly. The
tubes were then heated for 5 min in a boiling water
bath to develop the color and allowed to stand until
the color was settled for 5 min. The absorbance of
the developed color was read at 797 nm. The
standard curve of KH2PO4 was always run together
with samples.
RESULTS AND DISCUSSION
Soil and plant analysis
Industrial activities in Ras Malaab area resulted in
significant increase of soil heavy metal
concentrations comparing with Wadi Thal soil (Table
1). This result is consistent with those of Khan et al.
(2000) and Ciemense (2001) who reported that large
areas of land are contaminated with heavy metals as
a result of urban activities and industry. The heavy
metal content of the soil samples of Ras Malaab
exceeded that established by Anonymous (1986) for
Al3+ and was very close to exceed the permissible
limits for Zn2+. Table 1 showed increased heavy
metal pollution of Ras Malaab soil, which was
reflected in increasing their level in the shoot of
both species (Table 2). Heavy metal contents of the
shoot of both species were significantly increased in
Ras Malaab area comparing with those of Wadi Thal,
and more so in Z. coccineum (Table 2). On the
percentage basis, the greatest increase was found in
Al3+, Zn2+ and Cu2+ in the shoot of both species
(Table 2). Previous published data indicated that soil
heavy metal pollution increased heavy metal
concentrations in different plant species (Sanders et
al. 1986, Devi and Prasad 1998, Sebastiani et al.
2004, Rai et al. 2004, Tohidi et al. 2009). It is thus
obvious that industrial activities polluted the soil of
Ras Malaab, which resulted in increased heavy metal
levels in the shoots of both Zygophyllum species
Lipid peroxidation
MDA concentration increased in the shoot of
both species grown in the polluted soil of Ras
Malaab (Table 3), the increase was greater in Z.
EurAsian Journal of BioSciences 6: 1-10 (2012) Morsy et al.

Table 1. Minerals and heavy metal concentrations of Ras Malaab (polluted) and Wadi Thal (unpolluted) soils.

Each value is the mean±S.E. of three replicates.

*indicates significant difference of the control at least at 5% level.
ND: not detected, possibly its level was under the spectrophotometer sensitivity.

album than Z. coccineum. Lipid peroxidation
observed in the current work is indicative of heavy
metal-induced oxidative stress in both species. Our
results are in agreement with previous reports
where lipid peroxidation increased under soil heavy
metal pollution in different plant species (Chaffai et
al. 2005, Chaffai et al. 2009, Janicka et al. 2008). The
results of lipid peroxidation were comparable with
those of reactive oxygen species scavengers
(antioxidant enzyme activities) reported in the
following section (Table 3), where the increase in the
antioxidant enzyme activities was greater in Z. album
than Z. coccineum, in order to alleviate the greater
oxidative stress in induced Z. album.
Antioxidant enzymes
Antioxidant enzymes are very good biochemical
markers of stress and increasing their activities
could be a potential for remediation (Rucinska et al.
1999, Mittler 2002, Zembala et al. 2010). Except CAT,
the activities of GPX, ASO, APX and SOD were
increased in both species (Table 3), more so with
SOD. On the percentage basis, the increase in the
antioxidant enzyme activities was greater in Z. album
than in Z. coccineum (Table 3) to scavenge greater
lipid peroxidation observed in Z. album. Similar
results have been reported in different plant
species, where oxidative stress-induced by heavy
metals was alleviated by increasing antioxidant
enzyme activities (Rucinska et al. 1999, Ciemense
2001, Candan and Tarhan 2003, Zembala et al. 2010).
Increased oxidative defense systems have been
reported to be correlated with tolerance to
different stress conditions, thus plants perform
normally under the adverse environment (Mittler
2002, Candan and Tarhan 2003, Zembala et al. 2010).
Decreased activity of CAT observed in the current
study was previously found in pea seedlings under
cadmium stress (Sandalio et al. 2001). It can be
concluded that soil heavy metal pollution enhanced
oxidative stress in both Zygophyllum species, and
both species combat the heavy metal stress via
production of scavenging systems.

Table 2. Shoot minerals and heavy metal concentrations of two Zygophyllum species grown in Ras Malaab (polluted) and
Wadi Thall (unpolluted).

Table 3. Effect of heavy metal pollution on anitioxidant enzyme activities and lipid peroxidation (MDA concentration) of
two Zygophyllum species grown in Ras Malaab (polluted) and Wadi Thall (unpolluted).

5
EurAsian Journal of BioSciences 6: 1-10 (2012) Morsy et al.

Plasma membrane lipids increased in Z. coccineum while it was decreased in Z.

Total lipids, total phospholipids, total glycolipids
and total sterols were significantly decreased in
both species grown in polluted soil, the effect was
more pronounced in Z. coccineum (Table 4). Previous
published results indicated that copper and
cadmium stress decreased the total lipids,
phospholipids, glycolipids and sterols in pea
(Quartacci et al. 2000) pepper (Jemal et al. 2000) and
cucumber (Janicka et al. 2008).
Remarkable changes in the root PM
phospholipids (Table 5) and free fatty acids (Table 6)
were found in plants grown in polluted area. A
significant increased in PE was observed in the two
Zygophyllum species with a reduction in the PG and
PI contents, whereas the increase in PC was
insignificant (Table 5). The response of PS to heavy
metal pollution was varied in both species: it
album. Heavy metal stress decreased PC/PE ratio in
both species, the decrease was greater in Z. album.
The reduction in PC/PE ratio may influence the
plasma membrane performance under pollution,
because PC and PG have membrane stabilizing
effect relative to PE (Thompson 1992, Mansour et al.
1994, Salama et al. 2007). Our finding supports
previous published data where copper and cadmium
stress increased PE while they reduced PG content in
Zea mays (Chaffai et al. 2005, Chaffai et al. 2009).
Increased PE might have an adaptive value to
counterbalance the membrane rigidifying impact of
increasing saturated fatty acids (Thompson 1992,
Mansour et al. 1994, 2002, Salama et al. 2007)
induced in the current study under heavy metal
stress (Table 6).
The root PM unsaturated fatty acids (18:1, 18:2

Table 4. Effect of heavy metal pollution on the root PM total lipids, total sterols, total glycolipids and total phospholipids
of two Zygophyllum species grown in Ras Malaab (polluted) and Wadi Thall (unpolluted).

Table 5. Effect of heavy metal pollution on the root PM phospholipid composition (nmol g-1) and PC/PE ratio of two
Zygophyllum species grown in Ras Malaab (polluted) and Wadi Thall (unpolluted).

Table 6. Effect of heavy metal pollution on the root PM fatty acid composition (mol %) and saturated/unsaturated ratio of
two Zygophyllum species grown in Ras Malaab (polluted) and Wadi Thall (unpolluted).

6
EurAsian Journal of BioSciences 6: 1-10 (2012) Morsy et al.

and 20:1) was reduced whereas the saturated fatty
acids (14:0, 16:0) was increased in both species
under soil pollution (Table 6). This change in PM
fatty acid abundance lead to increased sat/unsat
ratio in the PM of both species, the effect was more
pronounced in Z. album. A decrease in fatty acyl
unsaturation of the PM under heavy metal pollution
has been also reported in wheat (Quartacci et al.
2000), Brassica juncea (Nouairi et al. 2006) and
tomato (Ben Ammar et al. 2007). The effect of fatty
acid saturation may be counterbalanced by the non-
significant increase in the PM PC, since PC increases
membrane lamellar structure and stability
(Thompson 1992, Mansour et al. 1994, 2002, Nouairi
et al. 2006, Ben Ammar et al. 2007, Salama et al.
2007). An increase in the degree of fatty acid
saturation is a typical reaction of plant PM to
different environmental stresses. In addition, certain
changes in the membrane lipids were reported to be
an adaptive response to different environmental
stresses, in order to restore optimum physical
membrane properties (Devi and Prasad 1999,
Mansour et al. 1994, 2002, 2003, Mansour and
Salama 2004, Nouairi et al. 2006, Ben Ammar et al.
2007, Salama et al. 2007). This appears to be the case
in the current work, where some PM lipid changes
might maintain membrane integrity under heavy
metal stress.
In conclusion, industrial activities increased the
level of heavy metals in the soil of Ras Malaab, which
resulted in drastic increase of shoot heavy metals of
both species. Accumulated heavy metals induced
oxidative stress in both species. Both Zygophyllum
species alleviated oxidative stress via increased
antioxidant enzyme activities. The root PM lipid
composition of Zygophyllum species was altered in
response to heavy metal pollution. Some of the PM
lipid changes might be in the favorable direction to
restore optimum membrane properties and
functions. Increased antioxidant enzyme activities as
well as PM lipid alterations might thus enable both
Zygophyllum species to withstand the prevailing
stressful environment.

REFERENCES

Anonymous (1986) Council directive on the protection of the environment, and in particular of the soil, when sewage
sludge is used in agriculture. Official Journal of the European Community, L181/6.
Beak KH, Skinner DZ (2003) Alteration of antioxidant enzyme gene expression during cold acclimation of near-isogenic
wheat lines. Plant Science 165: 1221-1227. doi: 10.1016/S0168-9452(03)00329-7
Ben Ammar W, Nouairi I, Zorrouk M, Jemal F (2007) Cadmium stress induces changes in the lipid composition and
biosynthesis in tomato (Lycopersicon esculentum Mill.) leaves. Plant Growth Regulation 53: 75-85. doi:
10.1007/s10725-007-9203-1
Black CA (1965) Methods of soil analysis. Parts 1 & 2, American Society of Agronomy, Madison, Wisconsin.
Breckle SW, Kahle H (1991) Geobotany and ecology. Progress in Botany 52: 391-406.
Candan N, Tarhan L (2003) The correlation between antioxidant enzyme activities and lipid peroxidation levels in
Mentha pulegium organs grown in Ca2+, Mg2+, Cu2+, Zn2+ and Mn2+ stress conditions. Plant Science 163: 769-779.
doi: 10.1016/S0168-9452(03)00269-3
Chaffai R, Ali T, Ferjani E (2005) Comparative effects of copper and cadmium on growth and lipid content in maize
seedlings (Zea mays L.). Pakistan Journal of Biological Sciences 8 (4): 649-655. doi: 10.3923/pjbs.2005.649.655
Chaffai R, Seybon TN, Marzouk B, Ferjani E (2009) A comparative analysis of fatty acidcomposition of root and shoot
lipids in Zea mays under copper and cadmium stress. Acta Biologia Hungarian 60: 109-125. doi:
10.1556/ABIOL.60.2009.1.10
Chinoy JJ, Singh YD, Gurumurti K (1976) Colorimetric determination of ascorbic acid turnover in plants. Indian Journal
Plant Physiology 19: 122-30.
Ciemense S (2001) Molecular mechanisms of plant metal tolerance and homeostasis. Planta 212: 475-480.
Deinstrop EH, Weinheim W (2000) Applied thin layer chromatography. Scottish Crop Research Institute, Dundee.
Devi SR, Prasad MN (1998) Copper toxicity in Ceratophyllum demeresum, a free floating macrophyte: response of
antioxidant enzymes and antioxidants. Plant Science 138: 157-165.

7
EurAsian Journal of BioSciences 6: 1-10 (2012) Morsy et al.

Devi SR, Prasad MN (1999) Membrane lipid alterations in heavy metal exposed plants. In: Prasad MNV, Hagemeyer J
(eds), Heavy Metal Stress in Plants. From Molecules to Ecosystems, Springer, Berlin, 99-116.
Gara LD, Pinto MC, Tommasi F (2003) The antioxidant systems and reactive oxygen species during plant-pathogen
interaction. Plant Physiology and Biochemistry 41: 863-870.
Hammerschmidt R, Nucles EM, Kuc J (1982) Association of enhanced peroxidase activity with induced systemic
resistance of cucumber to Colletotrichum lagenarium. Physiology of Plant Pathology 20: 73-82. doi: 10.1016/0048-
4059(82)90025-X
Harwood JL (1995) Recent environmental concerns and lipid metabolism. In: Kader JC, Mazliak (eds), Plant lipid
Metabolism, Kluwer Academic publishers, Dordrecht, 361-368.
Jackson ML (1967) Soil chemical analysis. Prinitice-Hall of India Private, New Delhi, India.
Janicka R, Katarzyna K, Marek B, Grazyna K (2008) Response of plasma membrane H+-ATPase to heavy metal stress in
Cucumis sativus roots. Journal of Experimental Botany 59: 3721-3728. doi: 10.1093/jxb/ern219
Jemal F, Zarrouk M, Ghorbal MH (2000) Effect of cadmium on lipid composition of pepper. Biochemical Society
Transition 28: 907-910.
Kappus H (1985) Lipid peroxidation; Mechanisms, Analysis, Enzymology and Biological relevance. In: Sies H (ed),
Oxidative Stress, Academic press, London, 273-310.
Kates M (1972) Techniques of lipidology. Isolation, analysis and identification of lipids. In: Work TS, Work E (eds),
Laboratory Techniques in Biochemistry and Molecular Biology, North Holland Publ, Amesterdam, 347-390.
Khan AG, Kuek TM, Chaudhury TM, Khoo CS, Hayes WJ (2000) Role of plants, mycorrhizae and phytochelators in heavy
metal contaminated land remediation. Chemosphere 41: 197-203. doi: 10.1016/S0045-6535(99)00412-9
Mansour MMF, Van Hasselt PR, Kuiper PJC (1994) Plasma membrane lipid alterations induced by NaCl in winter wheat
roots. Physiologia Plantarum 92:473-478.
Mansour MMF, Salama KHA, Al-Mutawa MM, Abou Hadid AF (2002) Effect of NaCl and polyamines on plasma
membrane lipids of wheat roots. Biologia Plantarum 45: 235-239.
Mansour MMF, Salama KHA, Al-Mutawa MM (2003) Transport proteins and salt tolerance in plants. Plant Science 164:
891-900. doi: 10.1016/S0168-9452(03)00109-2
Mansour MMF, Salama KHA (2004) Cellular basis of salinity tolerance in plants. Environmental and Experimental
Botany 52: 113-122. doi: 10.1016/j.envexpbot.2004.01.009
Mittler R (2002) Oxidative stress, antioxidants, and stress tolerance. Trends in Plant Science 7: 405-410. doi:
10.1016/S1360-1385(02)02312-9
Nouairi I, Ben Ammar W, Ben Youssef N, Ben Miled D, Habib GM, Zarrouk M (2006) Comparative study of cadmium
effects on membrane lipid composition of Brassica juncea and Brassica napus leaves. Plant Science 170: 511-519.
doi: 10.1016/j.plantsci.2005.10.003
Quartacci MF, Pinzino C, Sgherri CLM, Dalla Vecchia F, NavariIzzo F (2000) Growth in excess copper induces changes in
the lipid composition and fluidity of PSII-enriched membranes in wheat. Physiologia Plantarum 108: 87-93. doi:
10.1034/j.1399-3054-2000.108001087.x
Quiles MJ, López NI (2004) Photoinhibition of photosystems I and II induced by exposure to high light intensity during
oat plant grown effects on the chloroplastic NADH dehydrogenase complex. Plant Science 166: 815-823.
Rai V, Vaypayee P, Singh SN, Mehrotra S (2004) Effect of chromium accumulation on photosynthetic pigments,
oxidative stress defense system, nitrate reduction, proline level and eugenol content of Ocimum tenuiflorum. Plant
Science 167: 1159-1164. doi: 10.1016/j.plantsci.2004.06.016
Richards LA (1954) Diagnosis and improvement of saline and alkali soils. U.S. Department Agriculture, Handbook no.
60.
Rucinska R, Waplak S, Gwozdz A E (1999) Free radical formation and activity of antioxidant enzymes in lupine roots
exposed to lead. Plant Physiology and Biochemistry 37: 187-194. doi: 10.1016/S0981-9428(99)80033-3
Salama KHA, Mansour MMF, Ali FZM, Abou Hadid AF (2007) NaCl-induced changes in plasma membrane lipids and
proteins of Zea mays L. cultivars differing in their response to salinity. Acta Physiologia Plantarum 29: 351-359. doi:
10.1007/s11738-007-0044-3

8
EurAsian Journal of BioSciences 6: 1-9 (2012) Morsy et al.

Sandalio LM, Dalura HC, Gomez M, Romero-Puertas MC, Rio LA (2001) Cadmium induced changes in the growth and
oxidative metabolism of pea plants. Journal of Experimental Botany 52: 2115-2126.
Sanders JR, McGrath SP, Adams TM (1986) Zinc, copper and nickel concentrations in ryegrass grown on sludge
contaminated soils of different pH. Journal of Food Science Agriculture 37: 961-968.
Sebastiani I, Scebba F, Tongttti R (2004) Heavy metal accumulation and growth responses in poplar clones Eridano
(Populus deltoides × maximowiczii) and I-214 (P. × euramericana) exposed to industrial waste. Environmental and
Experimental Botany 52: 79-86.
Tappel AL (1973) Lipid peroxidation damage to cell components. Federation Proceedings of Biochemistry 32: 1870-
1874.
Thompson GR (1992) The regulation of membrane lipid, Academic Press, Boca Raton, Florida.
Tohidi Z, Baghizadeh A, Enteshari S (2009) The effect of aluminum and phosphorus on Brassica napus. American
European Agriculture and Environmental Sciences 6: 137-142.
Vaidyanathan H, Sivakumar P, Chakrabarsty R, Thomas G (2003) Scavenging of reactive oxygen species in NaCl-
stressed rice (Oryza sativa L.) - differential response in salt tolerant and sensitive varieties. Plant Science 165: 1411-
1418. doi: 10.1016/j.plantsci.2003.08.005
Vangronsveld J, Clijsters H (1994) Toxic effects of metals. In: Farago ME (ed), Plants and the chemical elements.
Biochemistry, uptake, tolerance and toxicity, Verlagsgesellschaft, Weinheim, 150-177.
Zembala M, Filek M, Walas S, Mrowiec H, Kornaś A, Miszalski Z, Hartikainen H (2010) Effect of selenium on macro-and
microelement distribution and physiological parameters of rape and wheat seedlings exposed to cadmium stress.
Plant and Soil 329(1-2): 457-468. doi: 10.1007/s11104-009-0171-2
Zhang F, Wang Y, Lou Z, Dong J (2007) Effect of heavy metal stress on antioxidative enzymes and lipid peroxidation in
leaves and roots of two mangrove plant seedlings (Kandelia candel and Bruguiera gymnorrhiza). Chemosphere 67:
44-50. doi: 10.1016/j.chemosphere.2006.10.007
Zornoza P, Vazquez S, Esteban E, Fernandez-Pascual M, Carpena R (2002) Cadmium-stress in nodulated white lupin:
strategies to avoid toxicity. Plant Physiology Biochemistry 40: 103-107. doi: 10.1016/s0981(02)01464-x

9
 EurAsian Journal of BioSciences 6: 1-10 (2012) Morsy et al.

Ağır Metallerin, Zygophyllum Türlerinin Plazma Zarları ve Antioksidan Enzimleri

Üzerine Etkisi

Özet
Giriş: Ağır metaller, toprakta yüksek konsantrasyonlarda bulunduklarında, büyük çevresel kirleticilerdir ve bitkilerin
büyüme ve gelişimi üzerinde toksik etkileri vardır. Endüstriyel aktiviteler, çok geniş alanlarda ağır metallerin birikmesi
ve doğal bitki örtüsünün etkilenmesini netice vermektedir. Bu yüzden, bitkilerin ağır metallerin olumsuz etkileri ile
nasıl başa çıktıklarının anlaşılması çok büyük önem arzetmektdir.
Materyal ve Metot: İki farklı alan seçildi. Bunlardan biri, bir alçıtaşı fabrikası yakınlarında, diğeri ise fabrikadan 25 km
uzaklıktaydı. İki Zygophyllum türü (Z. album ve Z. coccineum) çalışma alanlarında doğal olarak yetişmekteydi. Topraktaki
ağır metal stresinin; sürgün ağır metal konsantrasyonu, lipid peroksidasyonu, antioksidan enzim aktiviteleri ve kök
plazma zarı (PM) lipid kompozisyonu üzerine etkileri analiz edildi.
Bulgular: Çevre kirliliğine maruz kalan alanda yetişen her iki türün sürgünlerinde, ağır metal konsantrasyonları ve lipid
peroksidasyonu arttı. Kirlilik şartları altında, her iki türde de; askorbat oksidaz (ASO), gayakol (GPX), askorbat
peroksidaz (APX) ve süperoksit dismutaz (SOD) aktiviteleri artarken, katalaz (CAT) aktiviteleri azaldı. Z. album ve Z.
coccineum'da PM toplam lipidleri, fosfolipidler, glikolipidler ve steroller azaldı. Ağır metal stresi; her iki türün kök
PM'ında fosfatidiletanolamini (PE) artırdı, fosfatidilinositol (PI) ve fosfatidilgliserolü (PG) azalttı, fosfatidilkolinde (PC) ise
önemli bir değişiklik oluşturmadı. Fosfatidilserin; kirlilik koşullarında, Z. album PM'ında (PS) azalırken, Z. coccineum
PM'ında arttı. Ağır metal stresi, kök PM'ındaki yağ asitlerinin kompozisyon ve konsantrasyonunu değiştirdi ve her iki
türde doymuş/doymamış oranının değişmesine yol açtı.
Sonuç: Zygophyllum türlerinin ağır metal stresine dayanabilmesi için, etkin bir antioksidan mekanizmasının ve olumlu
PM lipid dengesinin önemli olduğunu göstermektedir.

Anahtar Kelimeler: Ağır metal, antioksidan enzim, lipid peroksidasyonu, plazma zarı yagı, yağ asidi, Zygophyllum sp.

10

View publication stats