Journal of Environmental Management 249 (2019) 109408

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Journal of Environmental Management

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Research article

Plant uptake and leaching potential upon application of amendments in soils T

spiked with heavy metals (Cd and Pb)
Iram Gula, Maria Manzoora, Imran Hashmia, Muhammad Faraz Bhattib, Jean Kallerhoffc,
⁎
Muhammad Arshada,
a
Institute of Environmental Sciences and Engineering (IESE), School of Civil and Environmental Engineering (SCEE), National University of Sciences and Technology
(NUST), Islamabad, Pakistan
b
Atta-ur-Rahman School of Applied Biosciences (ASAB), National University of Sciences and Technology (NUST), Islamabad, 44000, Pakistan
c
Ecolab, Université de Toulouse, CNRS, Toulouse, France

A R T I C LE I N FO A B S T R A C T

Keywords: The aim of this study was to assess the effects of soil amendments (organic and inorganic) on phytoavailability
Phytoextraction and leaching of cadmium (Cd) and lead (Pb) during enhanced phytoextraction. The vertical column study was
Soil amendments carried out to investigate the metal leaching meanwhile studying plant biomass and metals uptake for phy-
Leaching toextraction by Pelargonium hortroum. For this purpose, soil amendments at several concentration levels, such as
Cadmium
ethylene diaminetetraacetic acid (EDTA 0, 4, 5 mmol kg−1), ammonium nitrate (0, 8, 10 mmol kg−1), citric acid
Lead
(0, 8, 10 mmol kg−1), compost (0, 8%, 10%) and titanium dioxide nanoparticles (TNPs, 0, 80, 100 mg kg−1)
were used. Results revealed that EDTA efficiently improved Cd and Pb accumulation (mg kg−1) in shoots and
roots. Cd accumulation was significantly increased by 270%, 44%, 145%, and 6.4% in shoot and 94%, 19%, 48%
and 14% in root upon EDTA, ammonium nitrate, citric acid and TNPs application, respectively compared to the
control without amendment (WA). Similarly, Pb accumulation was significantly increased by 71%, 58% and 52%
in shoot and 88.8%, 70.6% and 64.6% in root upon exposure to higher levels of EDTA, citric acid and ammonium
nitrate, respectively when compared to the WA control. Application of TNPs reduced the Pb-accumulation in
shoots and roots by 33% and 28%, respectively. Similarly, the Pb-accumulation in shoots and roots was de-
creased by 39% and 35%, respectively upon compost addition. Plant biomass was significantly increased upon
application of soil amendments, with the exception of EDTA. The maximum Cd and Pb uptake was found in citric
acid amended soil. Leachate analysis revealed that the concentrations of Cd and Pb were increased by 109% and
101% in leachates upon EDTA application as compared to other amendments. In comparison with other
amendments, citric acid may be recommended as an environmentally friendly alternative for non-biodegradable
EDTA for enhanced phytoextraction of Cd and Pb.

1. Introduction
Soil polluted with heavy metals is a serious environmental issue.
Cadmium (Cd) and lead (Pb) contaminated soils are difficult to restore
due to the low mobility, persistent nature and high toxicity. These
heavy metals are naturally present in soils in trace amounts; higher Cd
and Pb soil concentrations are due to anthropogenic activities including
battery recycling industry, burning of coal, paints, automobile exhaust,
pesticides, leather tanning and mining (Manzoor et al., 2018a; Khan
et al., 2017; Arshad et al., 2008). Unlike organic contaminants, heavy
metals are non-biodegradable and remain in soil for long period of time.
Both heavy metals negatively affect plant biomass, photosynthetic
activity, stomata size, transpiration rate, nutrient uptake, ATPase ac-
tivity, necrosis and chlorosis (Khalid et al., 2017) and soil micro-
organisms by decreasing number of microbes, dehydrogenase activities,
respiration and microbial biomass (Manzoor et al., 2018b). Therefore,
remediating Cd and Pb contaminated soils and reducing the toxicity are
essential for environmental and human safety.
In recent years, phytoremediation–the use of living organisms
mainly plants for restoration of contaminated soil, have been widely
studied (Arshad et al., 2016). It is an environment friendly, esthetically
pleasant, economically feasible technique and has high acceptability
(Han et al., 2018; Manzoor et al., 2019). Phytoremediation is a broad
term which include various mechanisms based on plants’ ability to

⁎
Corresponding author.
E-mail address: marshad@iese.nust.edu.pk (M. Arshad).

https://doi.org/10.1016/j.jenvman.2019.109408
Received 24 February 2019; Received in revised form 11 August 2019; Accepted 14 August 2019
0301-4797/ © 2019 Elsevier Ltd. All rights reserved.
I. Gul, et al.
remove, degrade or immobilize contaminants. Phytoextraction (sub-
type of phytoremediation) is the green technology used for the removal
of heavy metals e.g. Cd and Pb from soil by using plants which have
capability to accumulate and translocate heavy metals to the aerial
plant parts (Chhajro et al., 2016). In this technique, heavy metals are
permanently removed from the soil and accumulated in the above
ground parts which is easy to handle, treat and dispose of. The profi-
ciency of phytoextraction depends on: a) capability of plant to survive
and accumulate Cd and Pb; b) available amount of these heavy metals
in soil (Dede and Ozdemi, 2016).
Chelate-assisted phytoextraction is an effective approach for re-
moval of Cd and Pb from soil by using plants (Zhang et al., 2016). EDTA
has been widely used for enhanced phytoextraction of Cd and Pb. The
application of EDTA showed increase in the solubility and accumulation
of Cd and Pb in shoots (Shahid et al., 2014). Cd and Pb concentrations
in Pelargonium hortorum were increased by 50% and 270% upon ap-
plication of 4 and 5 mmol kg−1 EDTA, respectively (Gul et al., 2019a).
It is well known that EDTA and its metal complexes are non-biode-
gradable, therefore, remain in soil for longer periods of time and pose
negative effects to plant biomass (Shahid et al., 2014). Other amend-
ments like citric acid (2 mmol kg−1) increased shoot biomass, root
biomass, chlorophyll a, tolerance index and Cd accumulation in Iris
halophila by 43%, 52%, 5%, 11%, and 15%, respectively (Han et al.,
2018). Organic and inorganic fertilizers are commonly used as additive
to provide nutrients for increasing crop growth and yield. Further the
N-containing fertilizers acidify the soil pH and increase the phytoa-
vailability of Cd and Pb. The application of ammonium nitrate in-
creased Cd and Pb accumulation and plant biomass (Ortiz et al., 2006).
Recently, titanium dioxide nanoparticles (TNPs) have been used for
enhancing nutrients uptake by crops and removal of heavy metals from
the contaminated medium (Zahra et al., 2017; Harikumar and Megha,
2017; Rafique et al., 2018).
In our previous work, application of these soil amendments (citric
acid, titanium dioxide nanoparticles (TNPs), ammonium nitrate, com-
post and EDTA) showed positive effect on enhancing the phytoaccu-
mulation of Cd in shoots of P. hortorum (Gul et al., 2018, 2019b). But,
the impact of these soil amendments on leaching and subsequent
ground water contamination is still not explored. Under this scenario,
there is a need to investigate such soil amendments which increase
phytoextraction of Cd and Pb with minimum leaching potenial. Keeping
in view this concern, present study was designed to investigate the ef-
fects of citric acid, ammonium nitrate, compost, TNPs and EDTA for
enhancing phytoavailability of Cd and Pb and, subsequently, increase
metal phytoextraction by Pelargonium hortorum, with minimum
leaching of Cd and Pb.
2. Material and methods
2.1. Soil spiking and amendments
Soil of clay loam texture with 7.39 pH, 0.38% organic matter and
0.33 mS cm−1 electrical conductivity (EC) was used for experimental
work. The detailed characteristics of soil have been mentioned else-
where (Gul et al., 2019b). The prepared soil was spiked with salts (la-
boratory grade form Sigma Aldrich) of cadmium sulfate (CdSO4) and
lead nitrate [Pb(NO3)2] at levels of 100, 150 mg Cd kg−1 and 1000,
1500 mg Pb kg−1, respectively. Control soil, without addition of Cd and
Pb, was also used for comparison. Deionized water was added to
maintain 60% moisture level, mixed daily for 1 month for metal sta-
bilization and homogenization in soil.
Experimental design was a completely randomized factorial (3 le-
vels of Pb and Cd; 5 types and 3 levels of amendments; 8 leaching
periods), with three replications. Soil amendments such as: EDTA (0, 4,
5 mmol kg−1), ammonium nitrate, citric acid (0, 8, 10 mmol kg−1),
compost (0, 8%, 10%) and TNPs (0, 80, 100 mg kg−1) were used to
assess the influence of these amendments on Cd and Pb leaching and
2
Journal of Environmental Management 249 (2019) 109408
uptake by P. hortorum.
2.2. Vertical column preparation for leaching study
Experimental unit consisted of rigid PVC column (15 cm dia-
meter × 50 cm height) with a hole at the bottom. The filter paper was
set at the bottom to avoid soil loss and water drainage for leachate
collection. The lower 10 cm of column was filled with pebble, followed
by addition of uncontaminated soil (20 cm) and contaminated soil
(20 cm) serving as the top layer. Columns were set as soil profile layers
and each layer was homogenized (Gabos et al., 2009). Control column
(10 cm Pebbles and 40 cm uncontaminated soil) was also used for
comparison. Plastic pots were set at the bottom of each vertical column
for leachate collection (Fig. S1).
One seedling (one–month old having similar size) of P. hortorum was
transplanted in each column and irrigated regularly to maintain
moisture level (70%). After 15 days of seedling transplantation, soil
amendments were applied and mixed with the upper soil layer, satu-
rated with deionized water, and exposed for 150 days. Leachates were
collected on 15, 30, 45, 60, 75, 90, 105 and 120 days after amendments
application.
2.3. Plant and leachate analysis
Collected leachates were filtered and analyzed for Cd and Pb by
using atomic absorption spectroscopy (AAS; PerkinElmer 900T). After
collection of last leachate sample, plants were carefully removed from
each column, washed with deionized water, immersed in 0.01M HNO3
(to remove Cd and Pb content attached to plant surface) and rinsed with
deionized water. Fresh biomass was recorded, plants were oven dried
(65 °C; 48 h) for dry biomass estimation. Afterwards, plant material was
digested in a mixture of HClO4 and HNO3 (1:3) and digests were ana-
lyzed for Cd and Pb content by AAS (Lu, 2000).
Cd and Pb uptakes by plant were calculated using following for-
mula.
Metal uptake = [Metalshoot × DSW ] + [Metalroot × DRW ] Eq.1
where;
Metalshoot and Metalroot are the concentrations of Cd and Pb in shoot
and root, respectively; DSW and DRW are the dry weights of shoot and
root, respectively.
2.4. Statistical analysis
Data presented are the means of three replicates along with stan-
dard deviation. Significant difference was determined using analysis of
variance (ANOVA) and means were compared by least significant dif-
ference (LSD) test at p < 0.05 using Statistix (8.0) software.
3. Results and discussion
3.1. Cd accumulation in shoot and root
Accumulation of Cd in shoot and root of P. hortorum increased with
increasing Cd levels (100–150 mg Cd kg−1). The Cd accumulation in
shoots and roots were significantly increased by 29% and 17%, re-
spectively, at 150 mg Cd kg−1 as compared to 100 mg Cd kg−1 without
amendments’ application (Fig. 1). In another study, P. hortorum accu-
mulated > 100 mg Cd kg−1 in shoots and have translocation factor
(transfer of Cd from root to shoot) more than 1, therefore, considered as
Cd-hyperaccumulator (Gul el at. 2019a). In the present study, appli-
cation of some amendments further enhanced the Cd-accumulation in
shoots and roots. The maximum Cd accumulation was observed in
EDTA amended soils followed by citric acid, ammonium nitrate, TNPs
and compost. The higher Cd accumulation in shoots (810 mg kg−1) and
roots (286 mg kg−1) was observed in EDTA (4 mmol kg−1) amended
I. Gul, et al. Journal of Environmental Management 249 (2019) 109408

Fig. 1. Cd concentrations in shoot and root of P. hortorum in spiked and
amended soil. Values presented in figure are the mean of three replicates with
standard deviation. The alphabets show the least significant difference
(p > 0.05). WA = Without Amendment; EDTA = Ethylene Diamine-
Tetraacetic Acid; CA = Citric Acid; AN = Ammonium Nitrate;
TNPs = Titanium Dioxide Nanoparticles; CO = Compost; A = Shoot; B = Root;
( = 100; = 150 mg Cd kg−1).
Fig. 2. Pb concentrations in shoot and root of P. hortorum in spiked and
amended soils. Values presented in figure are the mean of three replicates with
standard deviation. Different alphabets on the bars show the least significant
difference (p > 0.05). WA = Without Amendment; EDTA = Ethylene
Diamine-Tetraacetic Acid; CA = Citric Acid; AN = Ammonium Nitrate;
TNPs = Titanium Dioxide Nanoparticles; CO = Compost; A = Shoot; B = Root;
( = 1000; = 1500 mg Pb kg−1).

soils. Application of compost (10%) showed the minimum accumula- matter content which ultimately reduced the bioavailable Cd in soil by
tion of Cd in shoots (92 mg Cd kg−1) and roots (86.7 mg Cd kg−1). formation of organo-metallic complexes (Wiszniewska et al., 2016).
Upon exposure to 150 mg Cd kg−1, Cd accumulation was significantly
increased by 270%, 145% in shoots and 94%, 48% in roots with EDTA
3.2. Pb accumulation in shoot and root
(4 mmol kg−1) and citric acid (10 mmol kg−1), respectively. Applica-
tion of these soil amendments increased the bioavailability of Cd in soil
Fig. 2 illustrates Pb concentration in shoots and roots of P. hortorum.
which ultimately increased the phytoaccumulation of Cd. These results
Accumulation of Pb in shoots and roots ranged between 538 and
are in accordance to our previous work (Gul et al., 2018) where the
1817 mg kg−1 and 224–757 mg kg−1, respectively. At the highest Pb
application of EDTA, citric acid and TNPs enhanced Cd accumulation
treatment i.e. 1500 mg kg−1, Pb-accumulation in shoots and roots was
by 4.2–, 2.3– and 1.3–folds, respectively at 150 mg kg−1 treatment as
significantly increased by 28% and 15%, respectively, as compared to
compared to 25 mg kg−1. The variation in results of both studies are
1000 mg Pb kg−1 treatment without application of amendments. Ac-
due to difference in exposure time and Cd levels. In another study by
cumulation of Pb in plant might be due to the inherent ability of plant
Zhang et al. (2016), application of EDTA increased the Cd accumulation
to acidify soil pH by releasing H+ ions in soil, which is an important
by 53% in roots of castor cultivar (Ricinus communis L.). The increase in
factor in the accumulation of heavy metals (Arshad et al., 2016).
Cd accumulation upon EDTA application could be due to strong che-
Contrasting results were observed in Pb accumulation upon
mical stability of EDTA. In a recent study, it has been reported that
amendments’ application. TNPs increased Cd but decreased Pb accu-
Festuca arundinacea and Poa pratensis accumulated 3– and 1.7–folds
mulation in plants, probably due to different affinity for metal com-
more Cd in roots, respectively upon citric acid application (Wang et al.,
plexation with the two metals. EDTA, citric acid and ammonium nitrate
2017). This increase might be due to the role of citric acid in acidifying
application increased Pb accumulation. Maximum Pb-accumulation in
the rhizosphere soil and lowering the soil pH which could increase the
shoots (1817 mg kg−1) and roots (757 mg kg−1) was observed at
soluble Cd portion. In present study, application of ammonium nitrate
5 mmol kg−1 EDTA. In another study, accumulation of Pb in shoot was
(10 mmol kg−1) and TNPs (100 mg kg−1) increased the Cd accumula-
significantly increased by 51% upon 5 mmol kg−1 EDTA application
tion by 44%, 6.4% in shoots and 19%, 14% in roots, respectively at
(Gul et al., 2019a) over a period of 8 weeks. This increase in accumu-
150 mg Cd kg−1 treatment. The Cd accumulation in tobacco plant was
lation can be attributed to EDTA application resulting into decrease in
significantly increased when ammonium nitrate was applied (Ortiz
soil pH and increasing the bioavailability of Pb in soil. Application of
et al., 2006). This increase in Cd accumulation might be due to the role
EDTA significantly increased Pb accumulation in shoots and roots of
of N-forms of fertilizers i.e. NH4+ and NO3− which enhanced the root
Mirabilis jalapa by 1.7- and 45-folds, respectively (Yan et al., 2017). In
morphology and had high capability to get Cd. Furthermore, the pre-
the present study, minimum concentrations of Pb in shoots
sence of these forms of N increase the concentration of Cd in xylem sap
(538 mg kg−1) and roots (224 mg kg−1) were found at 10% compost
which enhanced the Cd accumulation in shoots. Cd accumulation in
application. Pb-accumulation was significantly increased by 71%, 58%
shoots of Eichhormia crassipes and Salvinia motesta was enhanced upon
and 52% in shoots and 88.8%, 70.6% and 64.6% in roots, upon ex-
TNPs application (Harikumar and Megha, 2017). In present study, ap-
posure to higher levels of EDTA (5 mmol kg−1), citric acid
plication of compost reduced the accumulation in shoots and roots by
(10 mmol kg−1) and ammonium nitrate (10 mmol kg−1), respectively.
52% and 41%, respectively. Addition of compost increases the organic
These amendments detached the Pb present in bound form by

3
I. Gul, et al. Journal of Environmental Management 249 (2019) 109408

acidifying soil pH, ultimately increasing the soluble portion of Pb which Table 2
could be available for plant uptake. Application of 100 mg kg−1 TNPs Pb uptake by P. hortorum in spiked soils upon amendments’ application.
significantly decreased the concentration of Pb in shoots by 33% and, in Pb uptake by P. hortorum in spiked soils (mg plant−1)
roots by 28% as compared to the same level of Pb (1500 mg kg−1).
Application of TNPs in anatase phase reduced Pb accumulation in shoot Pb treatment (mg kg-1) ► Control 1000 1500
and root of rice by 80% and 77%, respectively as compared to the Pb
Soil amendments▼
treatment alone (Cai et al., 2017). This reduction in the Pb accumula-
tion in rice tissue might by due to high sorption potential of nano- Without amendment 0 – 2.36 ± 0.24ghi 2.60 ± 0.26g
particles for Pb. The Pb-accumulation in shoots and roots was reduced EDTA (mmol kg−1) 4 – 3.48 ± 0.44f 3.61 ± 0.29ef
by 39% and 35%, respectively upon addition of 10% compost. Compost 5 – 3.37 ± 0.01f 3.49 ± 0.09f
Citric acid (mmol kg−1) 8 – 3.58 ± 0.18ef 4.24 ± 0.33cd
is an organic fertilizer, containing large amount of organic matter. The 10 – 4.28 ± 0.42c 5.05 ± 0.23a
presence of organic matter play an important role in the stabilization of Ammonium nitrate (mmol 8 – 3.49 ± 0.22f 4.37 ± 0.15bc
Pb. The Pb contents in soil get attached with the organic matter and Pb kg−1) 10 – 3.90 ± 0.58de 4.67 ± 0.32ab
becomes immobilized (Khan et al., 2017). Compost play significant role TNPs (mg kg−1) 80 – 2.21 ± 0.17hij 2.48 ± 0.30gh
100 – 1.95 ± 0.22jk 2.26 ± 0.14g-j
in stabilizing heavy metals rather than increasing the phytoavailability.
Compost (%) 8 – 1.94 ± 0.15jk 2.05 ± 0.05jk
This is also evident in the present study as the application of compost 10 – 1.83 ± 0.16k 2.06 ± 0.26ijk
decreased the Pb-accumulation in shoots and roots. By comparing all
amendments used in the present study, for enhanced phytoaccumula- Values presented in table are the mean of three replicates with ± standard
tion of Pb, EDTA was more efficient followed by other applied deviation values. Different alphabets show the least significant difference
amendments. (p > 0.05). EDTA = Ethylene Diamine-Tetraacetic Acid; TNPs = Titanium
Dioxide Nanoparticles.

3.3. Cd, Pb uptake and plant biomass

Plant dry biomass of P. hortorum exposed to Cd and Pb in combi-
nation with soil amendments is presented in Tables S1 and S2, re-
Cd and Pb uptake by plant without amendments’ application ranged
spectively. Application of EDTA in control group (no Pb and Cd) re-
from 0.58 to 0.66 mg and 2.36–2.60 mg per plant, respectively (Tables
duced the biomass by 24%. In a recent study, 24.8% reduction in shoot
1 and 2). The maximum uptake was observed at higher level of Cd
biomass of Phyllostachys pubescens was observed upon EDTA applica-
(150 mg kg−1) and Pb (1500 mg kg−1) in combination with citric acid
tion. This reduction in biomass might be due to the decrease in chlor-
(10 mmol kg−1); the minimum was observed upon 10% compost ap-
ophyll content because of EDTA-toxicity (Zhang et al., 2018).
plication.
In present study, plant dry biomass was decreased by 23% and 19%
Upon exposure to Cd (150 mg kg−1) and Pb (1500 mg kg−1), uptake
in Cd and Pb spiked soil without amendment application, respectively,
was significantly increased by 128% and 94%, respectively, when
as compared to the control. Gul et al. (2019a) reported that the biomass
10 mmol kg−1 citric acid was applied as compared to the same treat-
of P. hortorum was significantly reduced by 22.3% and 16.8% at
ment without amendments. A significant difference in Cd and Pb uptake
150 mg kg−1 Cd and 1500 mg kg−1 Pb. This decrease in biomass could
upon EDTA and citric acid application was observed. The accumulation
be attributed to heavy metal stress, increased membrane permeability
of Cd and Pb in citric acid amended soil was 31% and 10%, respec-
and oxidative stress. Zhang et al. (2018) reported that heavy metals
tively, lower than that with EDTA. However, biomass of plant in Cd and
have decreased viscosity and elasticity of cell wall and thereby, in-
Pb spiked soil in combination with citric acid was 51% and 58%, re-
hibited root growth.
spectively, higher than that with EDTA. Therefore, Cd and Pb uptake
Plant biomass was increased with increasing levels of citric acid,
was increased by 3.4% and 44.6%, respectively in citric acid amended
ammonium nitrate, TNPs and compost, but EDTA application decreased
soil as compared to EDTA at higher levels of Cd and Pb. Application of
the biomass of plants grown at the same level of Cd and Pb. Upon ex-
TNPs in Cd and Pb spiked soils reduced the Cd and Pb uptake by 41%
posure to higher level of Cd (150 mg kg−1), plant biomass was in-
and 55% respectively, as compared to citric acid amended soil. By
creased by 10.5%, 14.2%, 22.5%, and 23.6% in citric acid, ammonium
comparing all the amendments, citric acid was more efficient in terms
nitrate, TNPs and compost amended soils, respectively. Similarly, the
of Cd and Pb uptake as compared to the other amendments.
dry biomass of P. hortorum at 1500 mg Pb kg−1 showed an increase of
12.1%, 8.9%, 21.4% and 21.4% upon addition of citric acid, ammo-
Table 1
nium nitrate, TNPs and compost, respectively, as compared to the same
Cd uptake by P. hortorum in spiked soils upon amendments’ application.
treatment without amendments’ application. EDTA application sig-
Cd uptake by P. hortorum in spiked soils (mg plant−1) nificantly reduced dry biomass by 30.2% and 28.7% in Cd and Pb
spiked soil, respectively. TNPs are efficient in enhancing the biomass of
Cd treatment (mg kg-1) ► Control 100 150
crops and other plant species (Hanif et al., 2015; Zahra et al., 2017,
Soil amendments▼ 2019). This increase in biomass can be attributed to TNPs-enhanced
chlorophyll biosynthesis, enzymatic activities, increased photosynthesis
Without amendment 0 – 0.58 ± 0.05i 0.66 ± 0.05h
and nutrient uptake (Lyu et al., 2017; Rafique et al., 2018; Rizwan
EDTA (mmol kg−1) 4 – 1.47 ± 0.07a 1.45 ± 0.08ab
5 – 0.50 ± 0.02j 0.74 ± 0.06g et al., 2019). The TNPs application increased the concentration of
Citric acid (mmol kg−1) 8 – 1.24 ± 0.05c 1.39 ± 0.09b chlorophyll a and b as well as total chlorophyll in wheat (Kovacik et al.,
10 – 1.51 ± 0.06a 1.51 ± 0.10a 2014). In another study, biomass of tobacco plant was significantly
Ammonium nitrate (mmol 8 – 0.86 ± 0.00ef 0.90 ± 0.04e increased upon ammonium nitrate application. Ammonium nitrate
kg−1) 10 – 1.00 ± 0.01d 1.02 ± 0.03d
TNPs (mg kg−1) 80 – 0.83 ± 0.03f 0.82 ± 0.04f
contains NH4+ and NO3− ions, both are available for plant uptake,
100 – 0.80 ± 0.04e 0.89 ± 0.06ef therefore, increased the biomass (Ortiz et al., 2006). In general, am-
Compost (%) 8 – 0.48 ± 0.03jk 0.50 ± 0.03jk monium nitrate is used as a fertilizer to increase the plant growth and
10 – 0.39 ± 0.04l 0.43 ± 0.03kl yield.
Values presented in table are the mean of three replicates with ± standard
deviation values. Different alphabets show the least significant difference 3.4. Concentration of Cd and Pb in leachate
(p > 0.05). EDTA = Ethylene Diamine-Tetraacetic Acid; TNPs = Titanium
Dioxide Nanoparticles. For selection of any amendment for enhanced phytoextraction

4
I. Gul, et al.
Fig. 3. Cd concentration in leachates collected from spiked soil columns at
different time intervals, affected by different amendments in comparison to the
Cd only control. Values are the means of three replicates with standard de-
viation. A = 100 mg Cd kg−1; B = 150 mg Cd kg−1; ♦= Cd; ⬛ = Ethylene
Diamine-Tetraacetic Acid; ▲ = Citric Acid; ✖ = Ammonium Nitrate;
▬ = Titanium Dioxide Nanoparticles; ● = Compost.
strategy, the impact on leaching of Cd and Pb is an important factor.
Significant results were obtained at higher levels of soil amendments
i.e. EDTA (5 mmol kg−1), citric acid (10 mmol kg−1), ammonium ni-
trate (10 mmol kg−1), TNPs (100 mg kg−1) and compost (10%).
Therefore, results of Cd and Pb concentration in leachate are shown
only at these levels and results of other levels of soil amendments are
not shown due to their negligible values.
Application of EDTA, citric acid and ammonium nitrate increased
the Cd and Pb concentration in collected leachates as compared to un-
amended Cd and Pb groups. Compost application decreased the con-
centration in leachate as it stabilized the Cd and Pb (Khan et al., 2017;
Abbas et al., 2018). Cd and Pb concentrations in leachates decreased
with every continuing leachate collection (Figs. 3 and 4). Similar trends
were found by Lu et al. (2017). They reported that the application of
EDTA significantly increased Cd and Pb concentration in leached water
and with continuous collection of leachates, concentrations were de-
creased. This increased concentration of Cd and Pb in leachate might be
due to the role of EDTA in acidifying soil pH, at which solubility of
heavy metals is increased.
Maximum Cd concentration (191 mg kg−1) was found at 150 mg Cd
kg treatment in combination with 5 mmol kg−1 EDTA at 45th day of
−1
the leachate collection. At 150 mg Cd kg−1, the Cd concentration in
leachates was significantly increased by 109% upon 5 mmol kg−1 EDTA
application at 45th day as compared to 15th day of EDTA application.
By comparing other amendments with EDTA at higher level of Cd
(150 mg kg−1), the concentration of Cd in leachate was decreased by
63%, 57%, 84% and 89% upon citric acid, ammonium nitrate, TNPs
and compost applications, respectively (Fig. 3).
Pb is more persistent element with very low bioavailability.
Maximum Pb concentration in leachate was observed on 60th day of
EDTA (5 mmol kg−1) application (Fig. 4). The Pb concentration in
leachates was reduced by 69%, 67%, 91% and 92% upon citric acid,
ammonium nitrate, TNPs and compost application, respectively as
compared to EDTA. EDTA plays an important role in enhancing the
bioavailability of Cd and Pb. But, the bioavailable portion is more than
the amount taken up by plants, therefore, certain fraction is leached out
5
Journal of Environmental Management 249 (2019) 109408
Fig. 4. Pb concentration in leachates collected from spiked soil columns at
different time intervals, affected by different amendments in comparison to the
Pb only control. Values are the means of three replicates with standard devia-
tion. A = 1000 mg Pb kg−1; B = 1500 mg Pb kg−1; ♦ = Pb; ⬛ = Ethylene
Diamine-Tetraacetic Acid; ▲ = Citric Acid; ✖ = Ammonium Nitrate;
▬ = Titanium Dioxide Nanoparticles; ● = Compost.
from the soil. Application of other soil amendments (citric acid, am-
monium nitrate) reduced the Cd and Pb concentration in leachates
when compared with EDTA. Citric acid and ammonium nitrate might
have detached the bound Cd and Pb content from soil by acidification
and making them available for plant uptake in such a way that the
soluble portion is taken up by plants and minimum amount is leached
down. On the other hand, application of TNPs and compost stabilized
the Pb in soil and thus it was not found in the leachates. From this
study, it is evident that application of TNPs and compost in Pb-spiked
soils reduced the accumulation in plants and in collected leachates,
because it stabilized the Pb in soil, potentially making more pre-
cipitated forms of Pb.
4. Conclusions
EDTA was found to be the most efficient in increasing the accu-
mulation of Cd and Pb in shoot and root but leaching risk was also
enhanced. Ammonium nitrate and citric acid increased the accumula-
tion of Cd and Pb in shoot but was less efficient than EDTA. Heavy
metal uptake per plant was maximum in citric acid amended soil due to
higher biomass production with minimal leaching problem. By com-
paring all the amendments, citric acid can be recommended for en-
hanced phytoextraction of Cd and Pb contaminated soils and it could
serve as an environmentally friendly amendment.
Conflict of interest
The authors have no conflict of interest to declare related to the
reported work.
Acknowledgments
This study was the part of Pak-French collaborative project PHC-
PERIDOT jointly funded by the Higher Education Commission of
Pakistan (#1-1/PERIDOT/R&D/HEC/2015) and Campus France
(Project# 34362TM). This work was also supported by National
University of Sciences and Technology, Islamabad, Pakistan through
I. Gul, et al.
Mega S&T fund.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://
doi.org/10.1016/j.jenvman.2019.109408.
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