This article was downloaded by: [T&F Internal Users], [Mr Joel Peters]

On: 09 September 2014, At: 04:28

Publisher: Taylor & Francis
Informa Ltd Registered in England and Wales Registered Number: 1072954 Registered
office: Mortimer House, 37-41 Mortimer Street, London W1T 3JH, UK

Communications in Soil Science and

Plant Analysis
Publication details, including instructions for authors and
subscription information:
http://www.tandfonline.com/loi/lcss20

Effects of Exogenous Silicon on Cadmium

Accumulation and Biological Responses
of Nigella sativa L. (Black Cumin)
ab

Javad Sharifi Rad , Majid Sharifi Rad & Jaime A. Teixeira da Silva
a

Zabol Medicinal Plants Research Center, Zabol University of

Medical Sciences, Zabol, Iran
b

Department of Pharmacognosy, Faculty of Pharmacy, Zabol

University of Medical Sciences, Zabol, Iran
c

Department of Range and Watershed Management, Faculty of

Natural Resources, University of Zabol, Iran
d

Faculty of Agriculture, Kagawa University, Ikenobe, Kagawa, Japan

(retired)
Accepted author version posted online: 13 May 2014.Published
online: 21 Jul 2014.

To cite this article: Javad Sharifi Rad, Majid Sharifi Rad & Jaime A. Teixeira da Silva (2014)
Effects of Exogenous Silicon on Cadmium Accumulation and Biological Responses of Nigella sativa
L. (Black Cumin), Communications in Soil Science and Plant Analysis, 45:14, 1918-1933, DOI:
10.1080/00103624.2014.909835
To link to this article: http://dx.doi.org/10.1080/00103624.2014.909835

PLEASE SCROLL DOWN FOR ARTICLE

Taylor & Francis makes every effort to ensure the accuracy of all the information (the
Content) contained in the publications on our platform. However, Taylor & Francis,
our agents, and our licensors make no representations or warranties whatsoever as to
the accuracy, completeness, or suitability for any purpose of the Content. Any opinions
and views expressed in this publication are the opinions and views of the authors,
and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content
should not be relied upon and should be independently verified with primary sources
of information. Taylor and Francis shall not be liable for any losses, actions, claims,
proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or
howsoever caused arising directly or indirectly in connection with, in relation to or arising
out of the use of the Content.

Downloaded by [T&F Internal Users], [Mr Joel Peters] at 04:28 09 September 2014

This article may be used for research, teaching, and private study purposes. Any
substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing,
systematic supply, or distribution in any form to anyone is expressly forbidden. Terms &
Conditions of access and use can be found at http://www.tandfonline.com/page/termsand-conditions

Communications in Soil Science and Plant Analysis, 45:19181933, 2014

Copyright Taylor & Francis Group, LLC
ISSN: 0010-3624 print / 1532-2416 online
DOI: 10.1080/00103624.2014.909835

Downloaded by [T&F Internal Users], [Mr Joel Peters] at 04:28 09 September 2014

Effects of Exogenous Silicon on Cadmium

Accumulation and Biological Responses
of Nigella sativa L. (Black Cumin)
JAVAD SHARIFI RAD,1,2 MAJID SHARIFI RAD,3
AND JAIME A. TEIXEIRA DA SILVA4
1

Zabol Medicinal Plants Research Center, Zabol University of Medical Sciences,

Zabol, Iran
2
Department of Pharmacognosy, Faculty of Pharmacy, Zabol University of
Medical Sciences, Zabol, Iran
3
Department of Range and Watershed Management, Faculty of Natural
Resources, University of Zabol, Iran
4
Faculty of Agriculture, Kagawa University, Ikenobe, Kagawa, Japan (retired)
Medicinal plants are an age-old source of therapeutic agents to cure human disease.
Nigella sativa is used for edible and medicinal purposes in many countries. In this study,
biochemical and physiological responses of N. sativa to cadmium (Cd) toxicity were
investigated. Experiments were performed to study individual and combined effects of
exogenous silicon (Si) applied at 60, 120, and 180 mM in the form of sodium silicate
nonahydrate (Na2 SiO3 9H2 O) on Cd phytotoxicity in plants grown in perlite containing
different concentrations of cadmium nitrate (CdN2 O6 ). Cadmium treatment (120 M)
decreased chlorophyll and carotenoid content, dry weight, relative water content, and
root and shoot fresh weights compared with the control while proline content and lipid
peroxidation increased relative to the control. Black cumin is able to accumulate Cd, but
Si is also able to mitigate the negative consequences on growth and yield parameters.
Keywords
responses

Cadmium toxicity, medicinal plants, Nigella sativa, physiological

Introduction
Industrial pollution, specifically caused by heavy metals, and resistance of organisms
to heavy-metal toxicity have been widely studied over the past 30 years (Clements,
Palmgreen, and Kramer 2002; Lux et al. 2010; Ong et al. 2011; Tran and Popova 2013).
Pollution can be caused by increasing anthropogenic activities such as industrial and
municipal waste, ship-building activities, agricultural chemical toxin residues, municipal
solid waste, mining operations, and fertilizers that enter water bodies such as streams
and soil, having toxic effects on plants, animals, and soil and aquatic microorganisms
(MacFarlane and Burchett 2001). Knowledge of the interaction between plants and heavy
Received 3 April 2013; accepted 26 July 2013
Address correspondence to Majid Sharifi Rad, Department of Range and Watershed
Management, Faculty of Natural Resources, University of Zabol, Iran. E-mail: majid.sharifirad@
gmail.com

1918

Downloaded by [T&F Internal Users], [Mr Joel Peters] at 04:28 09 September 2014

Si Improved Cd Stress in Cumin

1919

metals in the environment can reduce the risks associated with the presence of heavy metals
in the food chain. Heavy metals enter the human diet from plants, and heavy-metal toxicity in humans includes damage to the nervous system, liver, kidneys, heart, blood vessels,
and bone tissue while inducing carcinogenesis and mutations (Sharma, Yelne, and Dennis
2005; Warrier, Nambiar, and Ramankutty 2004).
Cadmium (Cd), a nonessential element for plants, can affect plant growth and development. It is not a free metal in nature and often exists as a mineral combined with
other elements such as oxygen (cadmium oxide), chlorine (cadmium chloride), and sulfur (cadmium sulfide) (Clements, Palmgreen, and Kramer 2002). Cadmium is frequently
used in objects with metal coatings such as nickelCd batteries, color combinations, electronic components, and nuclear reactors (Adriano 2001). Cadmium is a strong pollutant
because of its severe toxicity at low concentrations and high solubility in water (Lone et al.
2008). Cadmium availability in soil is dependent on soil organic matter, root exudates,
mycorrhiza, soil pH, soil cation exchange capacity, temperature, soil temperature, and the
concentrations of other elements (Prasad et al. 2001). Unpolluted soil solutions contain
an estimated Cd concentration ranging from 0.04 to 0.32 mM (Wagner 1993). Cadmium
concentrations in soil solution varying from 0.32 to about 1 mM can be regarded as being
polluted to a moderate level (Sanit di Toppi and Gabbrielli 1999). Cadmium alters nutrient absorption by plants by competing with potassium (K), magnesium (Mg), calcium
(Ca), manganese (Mn), copper (Cu), zinc (Zn), and nickel (Ni) (Llamas, Ullrich, and Sanz
2000). Cadmium has a negative effect on plant metabolism, including the decrease in nutrient absorption and inhibited photosynthesis by affecting chlorophyll (chl) metabolism and
chloroplast structure, photosystem II activity, and enzymes controlling carbon metabolism
(Chaffei et al. 2004). Cadmium alters the structure and affects the function of membrane
lipids and membrane-associated enzyme activities, such as H+ ATPase (Obata, Inoue, and
Umebayashi 1996). Cadmium causes the stomata to close and reduces the amount of water
in the plant in the long term due to a reduction in plant growth and biomass. Another
toxicity-related aspect of Cd is the damage it causes to the nucleus and changes in RNA
synthesis (Liang, Wong, and Wei 2005). Cadmium toxicity causes oxidative stress due
to the production of free radicals and reduces the performance of antioxidant enzymes
in plants (Gallego, Benavides, and Tomaro 1996; Sandalio et al. 2001). At the molecular
level, Cd, which alters sulfhydryl oxidation, causes changes in protein secondary structure (Nazar et al. 2012). Cadmium is also involved in electron transport in chloroplasts
and mitochondria due to increased production of free radicals and damage to proteins,
lipids, and other biomolecules (Prasad et al. 2001; Shah et al. 2001). A plants response
to Cd toxicity lies within a complex network of physiological and molecular mechanisms
that include the maintenance and accumulation of metals in cell walls and root exudates
(Mohsenzadeh, Shahrtash, and Mohabatkar 2011).
Cadmium also chelates intracellular metals with organic acids, amino acids, ferritin,
phytochelatins, and metallothioneins and transfers them into the vacuoles (Hall 2002; Cho,
Chardonnens, and Dietz 2003). Cadmium also induces the synthesis of enzymatic and
nonenzymatic antioxidants in the immune systems biochemical responses (Hall 2002;
Cho, Chardonnens, and Dietz 2003).
Cadmium stress can be reduced in plants by altering soil nutrients, including the
addition of silicon (Si) to seeds before planting (Liang, Wong, and Wei 2005) or by exposing plants to mycorrhization (Schutzendubel and Polle 2002), salicylic acid (Karantev
2006; Popova et al. 2009), methyl jasmonate (Keramat 2009), or nitric oxide (Qiu et al.
2013).

Downloaded by [T&F Internal Users], [Mr Joel Peters] at 04:28 09 September 2014

1920

J. Sharifi Rad et al.

Silicon is the second most abundant element in the Earths crust and exists as silicon oxide or silicates between 0.1 and 0.6 mM in soil (Cahn 2009). Plants absorb Si as
silicic acid [Si(OH)4 ] (Epstein 1994). Although Si is not considered an essential element,
it has positive effects on growth, development, and productivity and its deposition protects plants and provides resistance to biotic and abiotic stresses in a range of plants (Ma
2004; Hodson et al. 2005; Ma and Yamaji 2006; Estelitano and Rodrigues 2012). Silicon
enhances plant tolerance to abiotic stresses, including those caused by heavy metals, and
the Si-mediated alleviation of heavy-metal toxicity in higher plants is widely accepted
(Iwasaki et al. 2002; Ma et al. 2002; Richmond and Sussman 2003; Shi et al. 2005; Cunha,
Nascimento, and Silva 2008; Mohsenzadeh, Shahrtash, and Mohabatkar 2011). However,
the underling mechanisms are still poorly understood. The consumption of plant material
with a high Cd content may cause toxicity in humans (FAO/WHO 1995). As a result of its
high toxicity, the maximum permissible limit of Cd in medicinal plants set by the World
Health Organization (WHO) is 0.3 g L1 .
Medicinal plants have long been a major source of therapeutic agents to cure human
disease. Nigella sativa Linn. (Ranunculaceae), commonly known as black seed or black
cumin, is cultivated in India, Syria, Lebanon, southern Europe, and Iran (Paarakh 2010).
Traditionally, N. sativa is used for edible and medicinal purposes in many countries. The
seeds are nutritionally and medicinally important as a pungent appetizer, an aromatic and
bitter stimulant, galactagogue, diuretic, anodyne, acrid deodorant, digestive, carminative,
febrifuge, purgative, abortifacent, and expectorant and are used to treat coughs, hydrophobia, fever, anorexia, dyspepsia, abdominal disorders, diarrhea, and internal hemorrhage
(Warrier, Nambiar, and Ramankutty 2004; Sharma, Yelne, and Dennis 2005; Paarakh
2010). Some medicinal plants have the potential to accumulate heavy metals such as Cd,
but the rate of uptake differs in each species based on their genetic characteristics (Peris
et al. 2007). Thus, the toxic effects of heavy metals in different plants may also differ significantly (Len et al. 2002). However, the ability of N. sativa to accumulate heavy metals
has not yet been reported in the literature. Thus, the focus of this study is the response of
N. sativa seedlings to Cd toxicity and their ability to reduce toxicity through the exogenous
application of Si.

Materials and Methods

Plant Materials and Treatments
Nigella sativa (cv. Baft) seeds were purchased from a seed company, Pakan Bazr (Esfahan,
Iran). Seeds were soaked in water for about 2 h, surface sterilized for 25 min with 10%
(w/v) sodium hypochloride solution, washed several times with deionized water, then
rinsed three times with sterile distilled water. Seeds were placed in Petri dishes containing
25 mL of deionized water. The containers were maintained for 3 days in the dark at laboratory temperature at 20 seeds Petri dish1 . Germinated seed (i.e., from which the epicotyl
and hypocotyl emerged) in Petri dishes were transferred to a growth chamber for 3 days and
then exposed to light with a fluorescent intensity of 5060 mol m2 s1 and a 16-h photoperiod, allowing seedlings to grow. Six-day-old seedlings were transferred to pots (1 kg
volume, 40 cm diameter) containing 0.5 kg of perlite (particle size: 37 mm, Tabandeh
Company, Khorasan, Iran) in a greenhouse at 25/18 C (day/night) and a 16-h photoperiod and covered with plastic to maintain 50% relative humidity. There were 20 seedlings
pot1 , which were irrigated once every 2 days with Hoaglands solution (Hoagland and
Arnon 1950). In total, seven different treatments were applied to the substrate, with three

Si Improved Cd Stress in Cumin

1921

replicates each, over a 20-day experiment as follows: control, Cd, Si, and Cd + Si. The
concentrations used were 120 M for Cd and 60, 120, and 180 mM for Si. Cadmium and
Si were added as cadmium nitrate tetrahydrate (CdN2 O6 4H2 O) and sodium silicate nonahydrate (Na2 SiO3 9H2 O), respectively. Fully developed leaves of 26-day-old plants were
used for biochemical analyses and shoot and root fresh weights (FW) were analyzed after
26 days and reported as g pot1 .

Downloaded by [T&F Internal Users], [Mr Joel Peters] at 04:28 09 September 2014

Fresh and Dry Weights of Seedling Shoots and Roots

The FW of 26-day-old seedling (50 pot1 ) shoots and roots was assessed for all treatments.
The dry weight (DW) of shoots and roots was determined after placing all plantlet organs
in an oven for 48 h at 75 C. In this experiment, shoot and root DW were expressed as
g pot1 , measured with 50 seedlings pot1 and three replicates treatment1 .
Measurement of Photosynthetic Pigments in Mature Leaves
Photosynthetic pigment content was measured by the method of Arnon (1959) using
200 mg of the oldest leaves of 26-day-old seedlings. Leaves were thoroughly ground in
80% acetone, made up to 25 mL with 80% acetone and then centrifuged at 4800 rpm for
20 min. The supernatant was used to measure the content of photosynthetic pigments. The
absorbance of the supernatant was determined by a Jenway model 6405 UV spectrophotometer (Jenway, Dunmow, UK) at 645 and 663 nm (Arnon 1959). The following formula
was used to estimate the chl a and b content of leaves:
mg chl g1 FW =

20.2 (OD645 nm ) + 8.02 (OD663 nm ) V

FW 100

where V = the final volume of solution in mL L1 and FW = the fresh weight of leaf tissue
in mg. Carotenoids were measured at 412, 431, 460, and 480 nm. The following formula
was used to estimate the carotenoid content of leaves:
-carotene (mg mL1 ) = 0.430 OD412 + 0.251 OD431
4.376 OD460 + 13.216 OD480
Measurement of Proline Content in Mature Leaves of N. sativa Seedlings
Proline (Pro) was extracted by the Bates, Waldren, and Teare (1973) method. Initially,
200 mg of 26-day-old seedling mature leaves derived from the control and all Cd and Si
treatments were weighed, and then 10 mL of 3% (w/v) sulfosalicylic acid (3-carboxy-4hydroxybenzene) was added. After 48 h, 2 mL of the solution was poured into a test tube
and 2 mL each of ninhydrin (2,2-dihydroxyindane-1,3-dione) and acetic acid were added.
Test tubes were heated in a hot water bath at 78 C for 1 h and then cooled rapidly on ice.
After cooling, 4 mL of toluene was placed in each tube and then vortexed for 20 s. After
settling, the red upper phase was used to measure Pro at 520 nm with a Jenway model
6405 UV spectrophotometer (Jenway, Dunmow, UK). Proline at 0, 40, 80, 120, 160, 200,
and 240 M was used to create a standard curve with the 0 M Pro test tube serving as the
blank.

1922

J. Sharifi Rad et al.

Downloaded by [T&F Internal Users], [Mr Joel Peters] at 04:28 09 September 2014

Measurement of Membrane Lipid Oxidation in Mature Leaves of N. sativa Seedlings

Membrane lipid peroxidation was measured by the Heath and Packer (1968) method based
on the formation of a malondialdehyde (MDA) complex. Leaf tissue (300 mg) of mature
26-day-old seedling leaves derived from the control and all Cd and Si treatments was
homogenized in 10 mL of 10% trichloroacetic acid (TCA; Sigma-Aldrich, St. Louis, Mo.,
USA) and then centrifuged for 15 min at 10,000 rpm. The seedlings used in this assay were
different to those used for the Pro assay. To 1 mL of the supernatant, 4 mL of TCA containing 25% of thiobarbituric acid (TBA; Sigma-Aldrich) was added and then vortexed,
and test tubes were placed in a boiling water bath (95 C) for 30 min and then rapidly
cooled in crushed ice. The absorbance of the cooled solution was determined at 532 nm by
subtracting the absorbance value of compounds at 600 nm.
Measurement of Cadmium in Mature Seedling Leaves and Roots of N. sativa
To measure the amount of Cd in roots and shoots, the Wickliff et al. (1980) method was
used. After seedlings were treated with different concentrations of cadmium nitrate, 26day-old plants were removed from Hoaglands solution and rinsed several times with
distilled water. After gently blotting off excess water around the roots, root and shoot
FW were measured separately. Plant samples were placed in an oven at 75 C for 48 h,
after which DW was measured; 0.3 g of dry shoots and 0.1 g of dry roots were placed in
high-temperature-resistant containers for 16 h at 500 C and ashed. Ashed shoot samples
were digested in 1.2 mL nitric acid (HNO3 ) (65%) for 4 h to which a further 1.8 mL of
HNO3 was added. Samples were further digested in 0.9 mL perchloric acid (HClO4 ) until a
clear solution was obtained. This solution was filtered and distilled water was added until a
final volume of 50 mL was obtained. To 0.4 g of ashed root samples, 0.6 mL of 65% nitric
acid was added and 0.3 mL of perchloric acid was added to digested samples until a clear
solution resulted. The final volume was brought to 21 mL using distilled water. To prepare
a standard curve, a stock solution was prepared with 0.5, 1, 1.5, 2, 2.5, and 10 mg L1
of cadmium nitrate Cd(NO3 )2 . Atomic absorption spectroscopy with an AA-220 (WFX130A model, Beijing, China) was used to measure the Cd content of standard root and
shoot solutions (Wickliff et al. 1980).
Measurement of Relative Water Content of Leaves of N. sativa Seedlings
To measure the relative water content (RWC), the fresh weight of N. sativa leaf segments
in control and stress samples were measured three times each. Leaf segments were placed
in distilled water in darkness for 24 h at 4 C to determine saturated weight (SW). The DW
was measured by placing samples in an oven for 24 h at 70 C. Then RWC was calculated
by the formula (Gonzlez and Gonzlez-Vilar 2001):
%RWC = [(DW FW) (DW SW)1 ] 100
Statistical Analysis
The experimental design was a randomized complete block with three replicates. One-way
analysis of variance (ANOVA) was calculated using SPSS v. 11.5 (IBM SPSS, New York,
USA) and differences between treatment means were compared using Duncans multiplerange test at < 0.05.

Si Improved Cd Stress in Cumin

1923

The FW (Figure 1) and DW (Figure 2) of shoots and roots of N. sativa seedlings increased
significantly in the presence of all Si concentrations relative to the control, even in the
presence of 120 M Cd. In all cases, the greatest concentration of Si applied (180 mM)
resulted in the greatest levels of FW and DW. A similar trend was observed for chl and
carotenoid content (Figure 3). The Pro content in fresh leaves was as low as the control
level when Si at any concentration was applied, but was significantly greater than the control when Cd alone was applied (Table 1). When Si was applied together with Cd, Pro levels
decreased significantly but never reached control levels, even when 180 mM was applied.
MDA content (i.e., level of lipid peroxidation) in leaves followed the exact same trend as
Pro (Table 1). The addition of Cd increased Cd content in shoots and roots, but the addition
of Si decreased Cd content in both organs significantly as the level of Si increased from
120 to 180 mM (Figure 4). The addition of Si alone significantly improved the RWC of
leaves (relative to the control), and the same trend was also observed when Cd was added
(Table 1).

Discussion
Effects of Cadmium and Silicon on Shoot and Root Fresh and Dry Weights
In this study, Cd significantly decreased the FW and DW of N. sativa shoots and roots
although the addition of Si helped to recover the losses caused by the negative impact
of Cd (Figures 1 and 2). Results showed that the FW of shoots and roots decreased during Cd stress more than the control, 65% and 39.5%, respectively. The root and shoot
DW under Cd stress decreased by 49% and 34% relative to the control. Cadmium stress
is caused by reduced water uptake, causing damage to microtubules and inhibiting cell
division, which in turn inhibits and decreases root growth (Eun, Youn, and Lee 2008).
Reduction in root growth could also be due to a decrease in cell wall elasticity in which
the middle blade calcium is replaced by Cd (Prasad 1995). Another possible reason for
5

Root fresh weigth

4.5

Shoot fresh weigth

c

3.5

f
g

3
g/pot

2.5

1.5
1

f
g

0.5

0
C

12
0

Si

18

0
Si
12
0
d
C

12
0

Si

12

60

0
12

0
18
Si

0
12
Si

60
Si

on

tro

Downloaded by [T&F Internal Users], [Mr Joel Peters] at 04:28 09 September 2014

Results

Figure 1. Effects of Si, Si + Cd, and Cd in the presence and absence of Cd (120 m) on Nigella
sativa seedling root and shoot fresh weight (FW). Different letters within root and shoot FW, assessed
separately, indicate significant differences (P < 0.05) according to Duncans multiple-range test.

1924

J. Sharifi Rad et al.

0.4

Root dry weigth

0.35

Shoot dry weight

a

0.3

g/pot

0.25

0.2

f
g

0.15
h

0.1
0.05

80

Si
1
+

Si

0
12
d
C

12

12

Si

12

60

0
12

80
Si
1

20
Si
1

60
Si

on
tro
l
C

Figure 2. Effects of Si, Si + Cd, and Cd in the presence and absence of Cd (120 m) on Nigella
sativa seedling root and shoot dry weight (DW). Different letters within root and shoot DW, assessed
separately, indicate significant differences (P < 0.05) according to Duncans multiple-range test.
6
5
mg/g fresh weight

Chlorophyll content

Carotenoid pigments

4
b
3

c
d

2
b

0
C

12

Si

18

0
Si
12
d
C

12

Si

12

60

0
12

0
18
Si

0
12
Si

60
Si

on

tro

Downloaded by [T&F Internal Users], [Mr Joel Peters] at 04:28 09 September 2014

Figure 3. Effects of Si, Si + Cd, and Cd in the presence and absence of Cd (120 m) on Nigella
sativa seedling chlorophyll (chl) content and carotenoid pigments. Different letters for chl and
carotenoid content, assessed separately, indicate significant differences (P < 0.05) according to
Duncans multiple-range test.

the reduction in root growth could be due to changes in membrane permeability and water
balance in plants (Barcelo et al. 1986). Cadmium inhibits cell division and the differentiation of cambium cells, resulting in a decrease in the number and diameter of these cells
(Barcelo and Poschenreider 1990). The reduction in water and nutrient movement may also
be due to the accumulation of wood lignin, phenols, and similar deposits of insoluble calcium oxalate (Furher 1982). Furthermore, Cd inhibits key enzymes of the photosynthetic
Calvin cycle, namely RUBP and PEP carboxylase, causes thylakoid membrane damage,
disrupts electron transport of iron and magnesium needed for photosystem (PS) II, and
decreases the photosynthesis and DW of Brassica juncea L. (brown mustard) roots and

1925

60

Si (mM)
120
180

Cd(M)
120

60 + 120

Notes. Values are expressed as means SE (n = 3). Different letters across treatments indicate significant differences (P < 0.05) according to Duncans multiplerange test.

78.33 0.66d
0.23 0.018d
0.03 0.003d

Si (mM) + Cd (M)
120 + 120
180 + 120

RWC 86.33 2.18c 92.33 1.66ab 95.66 0.33a 97.66 0.33a 64.66 2.40g 73.66 0.33def 76.33 1.20de
MDA 0.11 0.006e 0.11 0.003e 0.11 0.008e 0.12 0.005e 0.58 0.027a 0.40 0.026b
0.33 0.015c
Proline 0.01 0.003e 0.01 0.003e 0.01 0.003e 0.01 0.003e 0.1 0.000a
0.07 0.003b
0.05 0.003c

Control

Table 1
Effects of Si, Si + Cd, and Cd in the presence and absence of Cd (120 M) on RWC (%), MDA (mol/g fresh weight), and proline
(mol/g fresh leaf) of Nigella sativa seedlings

Downloaded by [T&F Internal Users], [Mr Joel Peters] at 04:28 09 September 2014

1926

J. Sharifi Rad et al.

Roots

Shoots
a

1
0.8

a
0.6

b
b

0.4

c
d

0.2
e

e
0
18
0
12
C

d
C

12

Si
+

+
0
12
d
C

Si

12

60
Si

12
d
C

on

tro

Downloaded by [T&F Internal Users], [Mr Joel Peters] at 04:28 09 September 2014

Cd content (mg/g dry weight)

1.2

Figure 4. Effects of Si, Si + Cd, and Cd in the presence and absence of Cd (120 M) on cadmium
content in roots and shoots of Nigella sativa seedlings. Different letters indicate significant differences (P < 0.05), assessed separately for roots and shoots, according to Duncans multiple-range
test.

shoots (Mohamed et al. 2012). Rodriguez et al. (1997) reported that the root and shoot
FW decreased in response to 0.05 mM of Cd stress in Zea mays L. (maize) and Pisum
sativum L. (pea). In this study, in the absence of Cd, root FW increased in the presence of
60, 120, and 180 mM Si by 31.2%, 43%, and 47.6%, respectively, and an increase in shoot
FW (11.7%, 18.3%, and 29.2%, respectively, compared to the control group) (Figure 1).
In plants treated with Si + 120 M Cd, the FW of roots and shoots increased compared to
plants stressed by 120 M Cd, such that 60 mM Si + 120 M Cd, 120 mM Si + 120 M
Cd, and 180 mM Si + 120 M Cd increased the FW of roots by 18%, 19.4%, and 41.2%
and the FW of shoots by 10.6%, 17% and 23.4%, respectively, compared to Cd-stressed
plants (Figure 1). In the plants treated with Si + 120 M Cd, 60 mM Si + 120 M Cd,
120 mM Si + 120 M Cd, and 180 mM Si + 120 M Cd, the DW of roots was increased
by 18%, 27.5%, and 29% and the DW of shoots by 11.1%, 16%, and 22.1%, respectively,
compared to Cd-stressed plants (Figure 2). Silicon at 5 mM stimulated the growth of young
Zea mays L. (maize) plants exposed to 50 M of Cd and influenced the development of
Casparian bands and suberin lamellae as well as vascular tissues in roots but did not affect
the distribution of apoplasmic and symplasmic Cd in roots, although there was a noticeable
decrease in symplasmic and increased apoplasmic concentration of Cd in shoots (Vaculk
et al. 2012). That study illustrated that the ability of Si to alleviate Cd toxicity might be
caused by intensified binding of Cd to the apoplasmic fraction in maize shoots. The DW of
Brassica oleracea L. (cabbage) shoots and roots declined sharply when exposed to 20, 50,
and 100 M Cd (Sun and Shen 2007). In contrast, Skrebsky et al. (2008) reported that in
a medicinal plant, Pfaffia glomerata (Spreng), the DW of both shoots and roots increased
significantly after exposure to 20 and 40 M Cd but was reduced in plants exposed to 80
M Cd. In contrast, root FW decreased significantly after exposure to Cd above 40 M.
Qiu et al. (2013) demonstrated that 150 M Cd significantly reduced plant height, root
length, and shoot and root FW and DW in wheat (Triticum aestivum L.) seedlings relative
to the control.

Si Improved Cd Stress in Cumin

1927

Downloaded by [T&F Internal Users], [Mr Joel Peters] at 04:28 09 September 2014

Effects of Cadmium and Silicon on Chlorophyll and Carotenoid Content

In this study, leaf chl and carotenoid levels decreased after exposure to stress caused
by 120 M Cd (55.2% and 49.8% compared to control) (Figure 3). There is a positive
correlation between carotenoid and chl concentration, and the loss of both is caused by
damage to membrane structures such as thylakoid membranes, changes in the structure of
proteins, inhibition of the electron transport chain in photosynthesis, and reduced PS II
and decreased absorption of minerals such as iron, magnesium, and manganese as metalbuilding components of chl (Bazzaz and Govindjee 1974; Alcntara et al. 1994; Rivetta,
Negrini, and Cocucci 1997). Reduced chl in older leaves may be due to the inhibitory
effect of Cd on nitrogen metabolism, which results in protein degradation in lower leaves
and transfers it to young leaves (Taiz and Zeiger 2002). Baryla et al. (2001) found that even
low concentrations of Cd in Brassica napus L. reduced chl content and photosynthetic reactions. Increasing Cd concentrations from 0 and 50 to 200 M negatively affected chl and
carotenoid content of B. juncea and activated the xanthophyll cycle, suggesting the need to
protect the photosynthetic apparatus from photoinhibition (Mohamed et al. 2012). Sun and
Shen (2007) reported that the net photosynthetic rate (P9, stomata1 conductance (Gs), photochemical efficiency of PS II (Fv Fm1 ), and quantum yield of electron flow through PS
II (phi (PS II)) in B. oleracea leaves declined sharply when exposed to 20, 50, and 100 m
Cd. Under the same level of Cd stress, the contents of chl a and b decreased, particularly
the latter, which could be an important factor inhibiting photosynthesis (Sun and Shen
2007). Skrebsky et al. (2008) reported that the chl content of P. glomerata was reduced
when Cd exceeded 40 M. In this study, following exposure to 60, 120, and 180 mM
of Si together with 120 M of Cd, the chl content increased by 23.1%, 40%, and 49%
and carotenoid content increased by 18%, 24.4%, and 29.9%, respectively, relative to Cdstressed plants (Figure 3). In the absence of Cd (i.e., 60, 120, and 180 mM Si alone), chl
contents increased 18%, 20%, and 24.3%, respectively, whereas the carotenoid contents
increased 29.2%, 32.5%, and 34.7%, respectively (Figure 3). This evidence confirms the
role of Si in assisting the recovery of losses caused by the negative impact of Cd. Dinakar
et al. (2008) evaluated the extent of damage to chl in leaves after 10 days of Cd stress (25,
50, and 100 L1 CdCl2 ): 100 L1 Cd decreased total chl in leaves by 91.01%, with
this loss increasing as Cd concentrations and exposure period increased.

Impact of Silicon and Cadmium on Proline Levels

In this study, in N. sativa, Pro increase 80% more than the control under stress caused by
120 M Cd (Table 1). This increase is likely due to be to the different roles of Pro as an
antioxidant and inhibitor of lipid peroxidation, as an osmolyte, as well as a metal receptor
regulating the pH of a cell (Siegenthaler et al. 1984). In this study, as Si concentration
increased in Cd-stressed plants, Pro decreased by 49.9% 54.6%, and 60.3%, when 60,
120, and 180 mM Si was applied (Table 1). Similar results have previously been reported
for Triticum aestivum (wheat) (Amani 2008), Phaseolus vulgaris L. (bean) (Zengin and
Munzuroglu 2005), and Raphanus sativus L. (radish) (Teklic et al. 2008). No new studies
on any other crop in the past 5 years have emerged. The accumulation of Pro in response
to an abiotic stress may be due to its increased de novo synthesis or decreased degradation
(Kasai et al. 1998). In heavy-metal stress, Pro accumulation as an osmolyte (Siegenthaler
et al. 1984), an acidity regulator in the cytosol (Shah et al. 2001), and a metal receptor
(Siegenthaler et al. 1984) plays an important role in reducing damage to membranes and
proteins. Therefore, the accumulation of Pro can be considered as an indicator of tolerance

1928

J. Sharifi Rad et al.

Downloaded by [T&F Internal Users], [Mr Joel Peters] at 04:28 09 September 2014

to heavy-metal stress. Dinakar et al. (2008) reported that Pro in leaves and roots of Arachis
hypogaea L. seedlings, evaluated after 10 days of 100 L1 Cd stress, increased 159.87%
more than the control.
Xu, Yin, and Li (2009) noted that Pro pretreatment reduced the level of reactive
oxygen species (ROS), protected the integrity of callus plasma membrane under Cd
stress, and therefore enhanced the tolerance of Solanum nigrum L. (common nightshade)
to Cd. In their study, inductively coupled plasmamass spectroscopy analysis showed
that exogenous Pro increased the accumulation of Cd in callus and regenerated shoots.
Improved Cd tolerance caused by Pro pretreatment was correlated with an increase of
superoxide dismutase and catalase activity and intracellular total glutathione content.
Effects of Cadmium and Silicon on Lipid Peroxidation in Leaves
MDA is one of the most frequently used indicators of lipid peroxidation. In this study, the
amount of MDA following treatment with 120 M Cd increased as much as 60% more than
the control (Table 1). This increase was likely due to oxidative damage in leaves caused by
Cd stress (Asada and Takashi 1987). As Si concentration increased, MDA levels decreased
in Cd-stressed N. sativa plants, such that Si at 60, 120, and 180 mM reduced MDA levels
by 19%, 44%, and 56.9%, respectively, compared to Cd-stressed plants.
Qiu et al. (2013) showed that 150 M Cd significantly enhanced the concentration of
MDA in wheat seedlings compared to the control. Amirjani (2012) reported that the MDA
content in wheat leaves exposed to Cd resulted in an accumulation of lipid peroxidation
products in leaves, but the accumulation was only significant when treated with 10 mg L1
Cd, in which MDA accumulated 63% more than the control.
Wheat plants growing in lead (Pb)contaminated (500, 1000, and 2500 M) soil significantly accumulated MDA content 1840% more than the control. Enhanced MDA
content suggested lipid peroxidation in response to Pb contamination and Pb {lead nitrate
[Pb(NO3 )2 ]}induced oxidative stress (Kaur et al. 2012). Keser and Saygideger (2010)
reported MDA accumulation in watercress (Nasturtium officinale) in response to Pb contamination, indicating that Pb-induced toxicity is exerted through the generation of free
radicals. Dey et al. (2007) suggested that even though total peroxide was not determined,
MDA levels increased with increasing Cd stress both in shoots and roots concomitant
with the metal concentration [cadmium chloride (CdCl2 ] (0200 M) and Pb(NO3 )2
(0200 M)), although, under Pb stress, the level of MDA only peaked at greater concentrations. They also suggested that this was probably because peroxidizable fatty acid
content became limiting. Therefore, an increase in MDA content suggests the prevalence
of oxidative stress and is a possible mechanism by which toxicity caused by Cd stress is
manifested in plants. Oxidative stress may have occurred due to changes in the activities
of antioxidative enzymes (Hasanuzzaman et al. 2012) although the prevalence of oxidative
stress can be confirmed by measuring the steady state levels of ROS in tissues.
Effects of Cadmium and Silicon on Cadmium Content in Roots and Shoots
In this study, the amount of Cd in roots and shoots increased 0.93 and 0.65 mg g1 DW
when N. sativa plants were exposed to 120 M Cd) Figure 4). Cadmium accumulated
more in the roots than in the shoots because Cd is more frequently deposited in roots than
in shoots (Blum 1997). Liang, Wong, and Wei (2005) noted that the roots of Zea mays
plants growing in soil contaminated with Cd had more Cd than shoots. In this study, Cd
decreased by 0.45, 0.3, and 0.17 mg DW in roots and by 0.53, 0.33, and 0.13 mg DW in

Si Improved Cd Stress in Cumin

1929

shoots, respectively, relative to only Cd-stressed plants, when 60, 120, and 180 mM of Si
was added to pots. Cunha, Nascimento, and Silva (2008) indicated that for maize plants
growing in Cd-contaminated soil, addition of Si caused Cd levels in roots and shoots to
increase.

Downloaded by [T&F Internal Users], [Mr Joel Peters] at 04:28 09 September 2014

Effects of Cadmium and Silicon on Leaf Relative Water Content

Relative water content (RWC) in N. sativa leaves decreased 10% compared to the control
under Cd stress (Table 1). Adding 60, 120, and 180 mM Si under Cd stress increased RWC
by 73%, 76%, and 78%, respectively, relative to the control (Table 1). Cadmium stress
can reduce water absorption due to reduced root growth and prevents the growth of hairy
roots due to microtubule damage, preventing cell division (Eun, Youn, and Lee 2008).
Because of a change in membrane permeability, water uptake and plant water balance
can also be reduced (Barcelo et al. 1986). Cadmium inhibits cell division and differentiation of cambium cells, resulting in a reduced number and diameter of cells and reduced
nutrients (Barcelo and Poschenreider 1990; Tran and Popova 2013). The reduction in structural changes in water and nutrient movement is also due to the accumulation of wood
lignin phenols and similar deposits of insoluble calcium oxalate (Furher 1982; Tran and
Popova 2013). Rodriguez et al. (1997) reported that RWC decreased in Zea mays and
Pisum sativum in response to 0.05 mM Cd.

Conclusion
Cadmium is one of the strongest environmental (mainly soil and air) heavy-metal pollutants. Cadmium uptake by plants is the first negative step in the contamination of the food
chain. Nigella sativa is useful in the treatment of human diseases, and thus its physiological
and biochemical responses to heavy-metal (Cd) stress were investigated. Cadmium stress
reduced growth parameters such as shoot and root DW and a decrease in chl, carotenoid,
and Pro content and an increase in lipid peroxidation (MDA content). The Cd concentration in shoots increased the level of accumulation in roots more than in shoots. Silicon,
when added as a plant nutrient amendment, reduced the negative effects of Cd stress and
decreased Cd accumulation in roots and shoots. Cadmium-containing phosphate fertilizers
used in agriculture are not recommended for the culture of N. sativa. However, if essential,
then they should be applied in the presence of Si.

References
Adriano, D. C. 2001. Trace elements in terrestrial environments: Biochemistry, bioavailability, and
the risks of metals, 2nd ed. New York: Springer.
Alcntara, E., F. J. Romera, M. Canete and M. de la Guardia. 1994. Effects of heavy metals on both
induction and function of root Fe (III) reductase in Fe-deficient cucumber (Cucumis sativus L.)
plants. Journal of Experimental Botany 45:18931898.
Amani, A. L. 2008. Cadmium-induced changes in pigment content, ion uptake, proline content, and
phosphoenolpyruvate carboxylase activity in Triticum aestivum seedlings. Australian Journal
of Basic and Applied Sciences 2 (1): 5762.
Amirjani, M. R. 2012. Effects of cadmium on wheat growth and some physiological factors.
International Journal of Forest, Soil, and Erosion 2 (1): 5058.
Arnon, D. I. 1959. Photosynthesis by isolated chloroplast, IV: Central concept and comparison of
three photochemical reactions. Biochimica et Biophysica Acta 20:440446.

Downloaded by [T&F Internal Users], [Mr Joel Peters] at 04:28 09 September 2014

1930

J. Sharifi Rad et al.

Asada, K., and M. Takahashi. 1987. Production and scavenging of active oxygen in photosynthesis.
In Photoinhibition, ed. D. J. Kyle, C. B. Osmond, and C. J. Arentzen, 227287. Amsterdam:
Elsevier.
Barcelo, J., C. Poschenreider, I. Andreu, and B. Gunse. 1986. Cadmium-induced decrease of water
stress resistance in bush bean plants (Phaseolus vulgaris L. cv. Contender). Plant Physiology
125:1725.
Barcelo, J., and C. Poschenrieder. 1990. Plant water relations as affected by heavy-metal stress: A
review. Journal of Plant Nutrition 13:137.
Baryla, A., P. Carrier, F. Frank, C. Coulomb, C. Sahut, and M. Havaux. 2001. Leaf chlorosis in
oilseed rape plants (Brassica napus) grown on cadmium polluted soil. Planta 212:696709.
Bates, L. S., R. P. Waldren, and I. D. Teare. 1973. Rapid determination of free proline for water stress
studies. Plant and Soil 39:205208.
Bazzaz, M. B., and Govindjee. 1974. Effects of cadmium nitrate on spectral characteristics and light
reactions of chloroplasts. Environmental Letters 6:112.
Blum, W. H. 1997. Cadmium uptake by higher plants. In Proceedings of extended abstracts from the
Fourth International Conference on the Biogeochemistry of Trace Elements, 109110. Berkeley:
University of California.
Cahn, R. W. 2009. Silicon: child and progenitor of revolution. In Into the Nano Era. Berlin: SpringerVerlag.
Chaffei, C., K. Pageau, A. Suzuki, H. Gouia, M. H. Ghorbel, and C. Masclaux-Daubresse. 2004.
Cadmium toxicity induced changes in nitrogen management in Lycopersicon esculentum leading to a metabolic safeguard through an amino acid storage strategy. Plant Cell Physiology
45:16811693.
Cho, M., A. N. Chardonnens, and K. J. Dietz. 2003. Differential heavy-metal tolerance of
Arabidopsis halleri and Arabidopsis thaliana: a leaf slice test. New Phytologist 158:287293.
Clements, S., M. G. Palmgreen, and U. Kramer. 2002. A long way ahead: Understanding and
engineering plant metal accumulation. Trends in Plant Science 7:309315.
Cunha, K., C. Nascimento, and A. Silva. 2008. Silicon alleviates the toxicity of cadmium and zinc for
maize (Zea mays L.) grown on a contaminated soil. Journal of Plant Nutrition and Soil Science
171:849853.
Dey, S. K., J. Dey, S. Patra, and D. Pothal. 2007. Changes in the antioxidative enzyme activities and
lipid peroxidation in wheat seedling exposed to cadmium and lead stress. Brazilian Journal of
Plant Physiology 19 (1): 5360.
Dinakar, N., P. C. Nagajyothi, S. Suresh, Y. Udaykiran, and T. Damodharam. 2008. Phytotoxicity of
cadmium on protein, proline, and antioxidant enzyme activities in growing Arachis hypogaea
L. seedlings. Journal of Environmental Sciences 20:199206.
Epstein, E. 1994. The anomaly of silicon in plant biology. Proceedings of the National Academy of
Sciences (USA) 91:1117.
Estelitano, M. E. M., and A. C. Rodrigues. 2012. Silicon location through backscattered electron
imaging and x-ray microanalysis in leaves of Cyperus ligularis L. and Rhynchospora aberrans
C. B. Clarke (Cyperaceae). Acta Botanica Brasilica 26:275280.
Eun, S. O., H. S. Youn, and Y. Lee. 2008. Lead disturbs microtubule organization in the root meristem
of Zea mays. Plant Physiology 110:357365.
FAO/WHO. 1995. Joint committee on food additives and contaminants (Position paper on cadmium,
prepared by France). 27th session. The Hague, the Netherlands: FAO.
Furher, J. 1982. Ethylene biosynthesis and cadmium toxicity in leaf tissue of beans (Phaseolus
vulgaris L.). Plant Physiology 70:162167.
Gallego, S. M., M. P. Benavides, and M. L. Tomaro. 1996. Effects of heavy-metal ions excess on
sunflower leaves: Evidence for involvement of oxidative stress. Plant Science 121:151159.
Gonzlez, L., and M. Gonzlez-Vilar. 2001. Determination of relative water content. In Handbook
of plant ecophysiology techniques, ed. M. J. R. Roger, 207212. Dordrecht, the Netherlands:
Kluwer Academic.

Downloaded by [T&F Internal Users], [Mr Joel Peters] at 04:28 09 September 2014

Si Improved Cd Stress in Cumin

1931

Hall, J. L. 2002. Cellular mechanism for heavy metal detoxification. Journal of Experimental Botany
53:111.
Hasanuzzaman, M., M. A Hossain, J. A. Teixeira da Silva, and M. Fujita. 2012. Plant response and
tolerance to abiotic oxidative stress: Antioxidant defense is a key factor. In Crop stress and
its management: Perspectives and strategies, ed. V. Bandi, A. K. Shanker, C. Shanker, and M.
Mandapaka, 261315. Dordrecht, the Netherlands: Springer.
Heath, R., and L. Packer. 1968. Photoperoxidation in isolated chloroplast, I: Kinetics and stoichiometry of fatty acid peroxidation. Archives of Biochemistry and Biophysics 125:189190.
Hoagland, D. R., and D. I. Arnon. 1950. The water culture method for growing plants without soil
(Circular 347). Berkeley: Agricultural Experiment Station, University of California.
Hodson, M. J. 2005. Phylogenetic variation in the silicon composition of plants. Annals of Botany
96:10271046.
Iwasaki, K., P. Maier, M. Fecht, and W. J. Horst. 2002. Leaf apoplastic silicon enhances manganese
tolerance of cowpea (Vigna unguiculata). Journal of Plant Physiology 159:167173.
Karantev, K., R. Yordanova, and L. Popova. 2006. Salicylic acid decrease cadmium toxicity in maize
plants. Plant Physiology (Special Issue): 4552.
Kasai, Y., M. Kato, J. Aoyama, and H. Hyodo. 1998. Ethylene production and increase in 1amino-cyclopropane-1-carboxylate oxidase activity during senescence of broccoli florets. Acta
Horticulturae 464:153157.
Kaur, G., H. P. Singh, D. R. Batish, and R. K. Kohli. 2012. Growth, photosynthetic activity,
and oxidative stress in wheat (Triticum aestivum) after exposure of lead to soil. Journal of
Environmental Biology 33:265269.
Keramat, B., M. Kalantari, and K. M. Arvin. 2009. Effects of methyl jasmonate in regulating
cadmium-induced oxidative stress in soybean plant (Glycine max L.). African Journal of
Microbiology Research 3:240244.
Keser, G., and S. Saygideger. 2010. Effects of lead on the activities of antioxidant enzymes in
watercress, Nasturtium officinale R. Br. Biological Trace Element Research 137:235243.
Len, A. M., J. M. Palma, F. J. Corpas, M. Gomez, M. C. Romero-Puertas, D. Chatterjee, R. M.
Mateos, L. A. del Ro, and L. M. Sandalio. 2002. Antioxidative enzymes in cultivars of pepper
plants with different sensitivity to cadmium. Plant Physiology and Biochemistry 40:813820.
Liang, Y., J. W. C. Wong, and L. Wei. 2005. Silicon-mediated enhancement of cadmium tolerance in
maize (Zea mays L.) grown in contaminated soil. Chemosphere 58:475483.
Llamas, A., C. I. Ullrich, and A. Sanz. 2000. Cd2+ effects on transmembrane electrical potential
difference, respiration, and membrane permeability of rice (Oryza sativa L.) roots. Plant and
Soil 219:2128.
Lone, M. I., Z. He, P. J. Stoffella, and X. Yang. 2008. Phytoremediation of heavy-metal-polluted
soils and water: Progresses and perspectives. Journal of Zheijang University Science B 9 (3):
210220.
Lux, A., M. Vaculk, M. Martinka, D. Likov, M. G. Kulkarni, W. A. Stirk, and J. van Staden.
2010. Cadmium induces hypodermal periderm formation in the roots of the monocotyledonous
medicinal plant Merwilla plumbea. Annals of Botany 10:18.
Ma, J. F. 2004. Role of silicon in enhancing the resistance of plants to biotic and abiotic stresses. Soil
Science and Plant Nutrition 50:1118.
Ma, J. F., K. Tamai, M. Ichii, and G. F. Wu. 2002. A rice mutant defective in Si uptake. Plant
Physiology 130:21112117.
Ma, J. F., and N. Yamaji. 2006. Silicon uptake and accumulation in higher plants. Trends in Plant
Science 11 (8): 2632.
MacFarlane, G. R., and M. D. Burchett. 2001. Photosynthetic pigments and peroxidase activity as
indicators of heavy metal stress in grey mangrove, Avicennia marina (Forsk.) Vierh. Marine
Pollution Bulletin 42:233240.
Mohamed, A. A., A. Castagna, A. Ranieri, and L. Sanit di Toppi. 2012. Cadmium tolerance
in Brassica juncea roots and shoots is affected by antioxidant status and phytochelatin
biosynthesis. Plant Physiology and Biochemistry 57:1522.

Downloaded by [T&F Internal Users], [Mr Joel Peters] at 04:28 09 September 2014

1932

J. Sharifi Rad et al.

Mohsenzadeh, S., M. Shahrtash, and H. Mohabatkar. 2011. Interactive effects of salicylic acid and
silicon on some physiological responses of cadmium-stressed maize seedlings. Iranian Journal
of Science and Technology Transaction A1: 5760.
Nazar, R., N. Iqbal, A. Masood, M. I. R. Khan, S. Syeed, and N. A. Khan. 2012. Cadmium toxicity
in plants and role of mineral nutrients in its alleviation. American Journal of Plant Sciences
3:14761489.
Obata, H., N. Inoue, and M. Umebayashi. 1996. Effect of Cd on plasma membrane ATPase from
plant roots differing in tolerance to Cd. Soil Science and Plant Nutrition 42:361366.
Ong, G. H., C. K. Yap, M. Maziah, and S. G. Tan. 2011. Heavy metal accumulation in a medicinal plants Centella asiatica from peninsular Malaysia. Journal of Biological Sciences 11 (2):
146155.
Paarakh, P. 2010. Nigella sativa Linn.: A comprehensive review. Indian Journal of Natural Products
and Resources 1 (4): 409429.
Peris, M., C. Mico, L. Recatala, R. Snchez, and J. Snchez. 2007. Heavy-metal contents in horticultural crops of a representative area of the European Mediterranean region. Science of the Total
Environment 378:4248.
Popova, L. P., L. T. Maslenkova, R. Y. Yordanova, A. P. Ivanova, A. P. Krantev, G. Szala, and T. Janda.
2009. Exogenous treatment with salicylic acid attenuates cadmium toxicity in pea seedlings.
Plant Physiology and Biochemistry 47:224231.
Prasad, M. N. V., P. Malec, A. Waloszek, M. Bojko, and K. Strzalka. 2001. Physiological responses
of Lemna trisulca L. (duckweed) to cadmium and copper bioaccumulation. Plant Science
161:881889.
Prasad, M. N. V. 1995. Cadmium toxicity and tolerance in vascular plants. Environmental and
Experimental Botany 35:525545.
Qiu, Z. B., J. L. Guo, M. M. Zhang, M. Y. Lei, and Z. L. Li. 2013. Nitric oxide acts as a signal molecule in microwave pretreatment induced cadmium tolerance in wheat seedlings. Acta
Physiologiae Plantarum 35 (1): 6573.
Richmond, K. E., and M. Sussman. 2003. Got silicon? The non-essential beneficial plant nutrient.
Current Opinion in Plant Biology 6:268272.
Rivetta, A., N. Negrini, and M. Cocucci. 1997. Involvement of Ca2+ -calmodulin in Cd2+ toxicity during the early phases of radish (Raphanus sativus L.) seed germination. Plant, Cell and
Environment 20:600608.
Rodriguez, E. L., L. E. Hernndez, P. Bonay, and R. O. Carpena-Ruiz. 1997. Distribution of cadmium in shoot and root tissues of maize and pea plants: Physiological disturbances. Journal of
Experimental Botany 48:123128.
Sandalio, L. M., H. C. Dalurzo, M. Gmez, M. C. Romero-Puertas, and L. A. Del Ro. 2001.
Cadmium-induced changes in the growth and oxidative metabolism of pea plants. Journal of
Experimental Botany 52 (364): 21152126.
Sanit di Toppi, L., and R. Gabbrielli. 1999. Response to cadmium in higher plants. Environmental
and Experimental Botany 41:105130.
Schutzendubel, A., and A. Polle. 2002. Plant responses to abiotic stresses: Heavy-metalinduced oxidative stress and protection by mycorrhization. Journal of Experimental Botany
53:13511365.
Shah, K., R. G. Kumar, S. Verma, and R. S. Dubey. 2001. Effect of cadmium on lipid peroxidation,
superoxide anion generation, and activities of antioxidant enzymes in growing rice seedlings.
Plant Science 161:11351141.
Sharma, P. C., M. B. Yelne, and T. J. Dennis. 2005. Database on medicinal plants used in Ayurveda.
New Delhi: CCRAS.
Shi, X., C. Zhang, H. Wang, and F. Zhang. 2005. Effect of Si on the distribution of Cd in rice
seedlings. Plant and Soil 272:5360.
Siegenthaler, U., U. Eicher, H. Oeschger, and W. Dansgaard. 1984. Lake sediments as continental
18 O records from the glacial/post-glacial transition. Annals of Glaciology 5:149152.

Downloaded by [T&F Internal Users], [Mr Joel Peters] at 04:28 09 September 2014

Si Improved Cd Stress in Cumin

1933

Skrebsky, E. C., L. A. Tabaldi, L. B. Pereira, R. Rauber, J. Maldaner, D. Cargnelutti, J. F.

Gonalves, G. Y. Castro, M. R. C. Shetinger, and F. T. Nicoloso. 2008. Effect of cadmium
on growth, micronutrient concentration, and -aminolevulinic acid dehydratase and acid phosphatase activities in plants of Pfaffia glomerata. Brazilian Journal of Plant Physiology 20 (4):
285294.
Sun, J. Y., and Z. G. Shen. 2007. Effects of Cd stress on photosynthetic characteristics and nutrient
uptake of cabbages with different Cd-tolerance. Ying Yong Sheng Tai Xue Bao 18:26052610.
Taiz, L., and E. Zeiger. 2002. Plant physiology, 3rd ed. Sunderland: Sinauer Associates, Inc.
Teklic, T., J. T. Hancock, M. Engler, N. Paradikovic, V. Cesar, H. Lepedus, I. tolfa, and D. Belo.
2008. Antioxidative responses in radish (Raphanus sativus L.) plants stressed by copper and
lead in nutrient solution and soil. Acta Biologica Cracoviensia Series Botanica 50:7986.
Tran, A. T., and L. P. Popova. 2013. Functions and toxicity of cadmium in plants: Recent advances
and future prospects. Turkish Journal of Botany 37:113.
Vaculk, M., T. Landberg, M. Greger, M. Luxov, M. Stolrikov, and A. Lux. 2012. Silicon modifies
root anatomy, and uptake and subcellular distribution of cadmium in young maize plants. Annals
of Botany 110 (2): 433443.
Wagner, G. J. 1993. Accumulation of cadmium in crop plants and its consequences to human health.
Advances in Agronomy 51:173212.
Warrier, P. K., V. P. K. Nambiar, and C. Ramankutty. 2004. Indian medicinal plants: A compendium
of 500 species. Chennai, India: Orient Longman.
Wickliff, C., H. J. Evans, K. R. Carter, and S. A. Russell. 1980. Cadmium effects on the nitrogen
fixation system of red alder. Journal of Environmental Quality 9:180184.
Xu, J., H. Yin, and X. Li. 2009. Protective effects of proline against cadmium toxicity in
micropropagated hyperaccumulator, Solanum nigrum L. Plant Cell Reports 28 (2): 325333.
Zengin, F. K., and O. Munzuroglu. 2005. Effects of some heavy metals on content of chlorophyll,
proline, and some antioxidant chemicals in bean (Phaseolus vulgaris L.) seedling. Acta
Biologica Cracoviensia Series Botanica 47:157164.