Archives of

1. Introduction
Today, there is a need to develop biological methods of soil amendment which are economical in terms of cost and eliminate pollution without reducing soil fertility [1]. Much research has been conducted in the field of biostimulation and bioavailability of heavy metals [2,3]. Recently, EDTA has been widely used to reduce the toxicity of heavy metals [4,5]. EDTA is a polyamine carboxylic acid with the chemical formula CH2N(CH2CO2H)2, which is mostly used as its disodium salt. This compound forms stable complexes with a wide range of metals, so it has a high ability to release metal ions [6]. Heavy elements are among the most important environmental pollutants, which have been highly considered in the last few decades [7]. The accumulation of elements in the soil, especially in agricultural lands, is a gradual thing, and the concentration of heavy elements can reach a level that threatens human food security [8].
Every year, thousands of tons of these elements, which are caused by urban, industrial, and agricultural activities, enter the soil [9]. Heavy metals have a very destructive effect on the environment and living organisms, including humans [9]. Their greatest effect on humans is related to neurological disorders [10,11]. These metals also replace other minerals needed by the body [12]. Also, the accumulation of heavy metals in the soil and water has received much attention due to their toxicity and the dangers they pose to humans and the environment [13]. Excessive accumulation of heavy metals in the soil destroys the ecosystem and affects soil properties such as pH, electrical conductivity, cation exchange capacity, and microbial and biological activities [14,15]. Many methods have been used to reduce soil pollution from heavy metals, which are classified into physical, chemical, and biological groups or a combination of these [16,17]. One of the effective compounds in the absorption of heavy metals is the use of EDTA [18]. In various research, the effect of EDTA in increasing the absorption of heavy metals has been investigated.
Dong et al stated that biological stimulation with EDTA and electrokinetic results in increasing the absorption of heavy elements by plants [19]. Cheng et al also stated that soil washing using organic EDTA has led to the removal of significant amounts of nickel, copper, and zinc
The Effect of EDTA on the Ability to Absorb Different Concentrations of Nickel, Cadmium, and Lead in Soil by Corn Plants
Ali Asnaashari1ID, Sima Sabzalipour1*ID, Kamran Mohseni Far2ID, Mojtaba Alavifazel3ID
1Department of Environmental Science, Ahvaz Branch, Islamic Azad University, Ahvaz, Iran
2Department of Soil Science, Ahvaz Branch, Islamic Azad University, Ahvaz, Iran
3Department of Agronomy, Ahvaz Branch, Islamic Azad University, Ahvaz, Iran
*Corresponding Author: Sima Sabzalipour, Email: shadi582@yahoo.com
Abstract
Background & Aims: The use of resistant and widely used species such as corn in agriculture and industry can be an effective solution for the bioremediation of soil pollutants, including heavy elements. The current research aimed to investigate the effect of EDTA on the ability to absorb different concentrations of heavy metals in the soil of corn plants in 2019.
Materials and Methods: This research was conducted as a three-factor pilot design at concentrations of 0, 50, and 100 mg/kg in the greenhouse. The samples were analyzed using an atomic absorption device. The laboratory pilot design was based on Taguchi’s algorithm. Finally, transfer factor (TF), bioconcentration factor (BCF), and bioaccumulation coefficient (BAC) were calculated.
Results: The results showed that different organs (root, stem, and leaf) show different levels of bioaccumulation under the influence of variable factors in different concentrations of heavy elements (nickel, cadmium, and lead). Also, there is a significant difference between the measured amounts of heavy elements in different organs of the corn plant (P ≤ 0.05). The average TF levels for elements at concentrations of 50 and 100 mg/kg were 0.79 and 1.66 for nickel, 0.82 and 0.78 for cadmium and 1.361, 1.378, and 1.387 for lead. Based on the results, with the increase in nickel concentration, the absorption level increased, and with the increase in cadmium concentration, the absorption level decreased.
Conclusion: The results of this research showed that it is possible to use EDTA to increase the efficiency of corn plants in absorbing heavy elements of nickel, cadmium, and lead.
Keywords: Bioaccumulation, Corn, Transfer factor, Heavy metals, EDTA
Received: Septmeber 16, 2022, Accepted: May 2, 2023, ePublished: September 29, 2023
https://jhygiene.muq.ac.ir/
10.34172/AHS.12.3.1.415
Vol. 12, No. 3, 2023, 148-154
Original Article
© 2023 The Author(s); This is an open-access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Arch Hyg Sci. Volume 12, Number 3, 2023 149
Effect of EDTA on heavy metals absorbtion in soil by Corn plants
in industrial soils [20]. Zhang et al showed the effect of
EDTA on increasing the absorption of heavy elements by
bamboo by more than 38% [21]. Hamidpour et al stated
that growth-promoting bacteria, along with EDTA, led
to an increase in the absorption of heavy elements by
biomass [22].
But the level of effectiveness of EDTA in absorbing
heavy elements is different based on the type of plant and
different elements [23-25]. The methods used to estimate
the absorption level of heavy elements by plants are
transfer factor (TF), bioconcentration factor (BCF), and
bioaccumulation coefficient (BAC) indices [26]. In these
indices, the ratio of the amount of heavy elements in the
soil to the amount of elements absorbed in the plant organs
is calculated [27]. Corn is one of the most widely used
crops in the southern regions of Iran [28]. Maize, with the
scientific name Zea mays, from the cereal category and
the large wheat family, is a monocotyledonous plant, and
its edible part is the female inflorescence (spike) that
produces seeds or the same fruit [29].
Nickel, cadmium, and lead are heavy elements that
are mainly rooted in industrial activities [30]. The
results of various studies have shown that the level of
soil contamination with heavy metals in Khuzestan
province is more than in other regions of Iran. The
results of Sahipour and Sabzalipour’s study showed that
the intensity of soil contamination with heavy elements
lead, zinc, nickel, and chromium is at the “contaminated”
level [31]. Shahidi Kaviani and Paykanpour Fard also
found that the contamination of the soils of the Ahvaz oil
field, especially in the vicinity of oil and gas wells, with
heavy metals cadmium and copper is higher than the
world average [32]. Considering the existence of many
industries such as oil, gas, and steel in the southwest of
Iran, as well as the areas under corn cultivation, in this
research (in 2019), the effect of EDTA on the ability to
absorb different concentrations of heavy metals in the soil
by corn plants has been studied.
2. Material and Methods
This research is based on the practical purpose and
descriptive data collection, which was conducted to
investigate the ability of grain corn species to absorb heavy
metals (nickel, cadmium, and lead) from soil using EDTA
in 2019. The research is a three-factor factorial experiment
in the form of a completely randomized design with three
replications in the greenhouse. Soil samples were collected
from the surrounding areas of Ahvaz city at a depth of 0-60
cm. After collection, the samples were dried and passed
through a 2 mm sieve before use [33]. The measurement
of nickel and cadmium elements was also done by an
Agilent 240 AA furnace atomic absorption device. In
the study of Guo et al, EDTA at the level of 12 mmol was
determined as the optimal limit of bioavailability of heavy
metals from the soil, so the amount of EDTA at the level
of 12 mmol/kg was used. Also, to investigate the effect
of pollution level on the absorption of heavy metals,
nickel concentration (zero, 50, 100 mg nickel/kg soil
as nickel sulfate) and cadmium concentration in three
levels (zero, 50, 100 mg cadmium/kg soil as cadmium
nitrate) and lead concentration in three levels (zero, 50,
100 mg lead/kg soil as lead nitrate) were used [23]. For
preparation, 3 kg of prepared dry soil was transferred to
the pots and uniformly contaminated with heavy metal
salt treatments of cadmium, nickel, and lead. After
about a month, EDTA treatments were added to the
pots. The pots were kept for 45 days to create balance
in the soil [34]. Then corn seeds were planted in each
pot in such a way that three corn plants were harvested
from each pot. For 12 weeks, daily, pots containing
corn plants were repeatedly visited and irrigated with
distilled water, low-quality plants were thinned in terms
of growth, and the edges of the pots were also removed
for the uniformity of the absorption conditions of the
plants. Also, to achieve uniform environmental growth
conditions, the pots were changed every week. After
about 100 days, when the plant achieved proper growth,
the plants were harvested, and after washing the roots,
three tissues of the root, stem, and leaf were separated
until the absorption of heavy metals was done by the
atomic absorption device. The method (Jackson 1958)
was used to digest plant samples [35].
After determining the amount of extractable heavy
metals in corn plant and soil samples, we measured the
TF index (transfer factor: ratio of metal concentration in
aerial parts of plants to metal concentration in roots), BCF
index (Bioconcentration: ratio of metal concentration
in plant roots to metal concentration in soil), and BAC
index (biological accumulation coefficient: ratio of
metal concentration in aerial parts of plants to metal
concentration in soil).
2.1. Transfer factor
This factor is used to evaluate the plant’s ability to transfer
metal from the root to the stem and is calculated by
dividing the concentration of the element in the aerial
part by the concentration of the element in the root [36].
Equation 1: Transfer factor calculation
TF = C(Cd ,Ni),sh / C(Cd,Ni),r Eq. (1)
2.2. Bioconcentration factor index
One of the important factors used to measure the
concentration of heavy elements in plant samples is the
bioconcentration factor, and it is calculated by dividing
the concentration of the element in the aerial part by the
concentration of the element in the soil [36].
Equation 2: Calculation of biological inhibition index
BCF = C(Cd,Ni),r/C(Cd ,Ni),s Eq. (2)
Asnaashari et al
150 Arch Hyg Sci. Volume 12, Number 3, 2023
2.3. Bioaccumulation coefficient
This index shows the ratio of metal accumulation in plant
roots to metal accumulation in soil. (accumulation in
roots to accumulation in soil) [37].
Equation 3: Calculation of bioaccumulation coefficient
BAC = C(Cd,Ni),Sh/C(C ,Ni),s Eq. (3)
3. Results
The results of measuring the heavy elements of nickelcadmium
in soil, root, stem, and leaf samples of corn
plants at concentrations of 0, 50, and 100 mg/kg and their
standard deviation are presented in Table 1.
The results showed that the amounts of nickel, cadmium,
and lead in the factor variable and biomass in unpolluted
pots were lower than the samples contaminated with
concentrations of 50 and 100 mg/kg (Figures 1, 2, and 3).
Also, the amounts of nickel, cadmium, and lead elements
in the soil samples were lower than in the corn biomass
samples (root, stem, and leaf) (Figure 2). The difference
between the amounts of heavy elements nickel and
cadmium in soil and plant samples was statistically proven
with 95% confidence. The highest amount of nickel in the
biomass of root samples at a concentration of 100 mg/kg
was measured as 5.79 mg/kg, and the lowest amount was
measured as 1.09 mg/kg in control pots (without heavy
elements). Also, the highest amount of lead in soil samples
was 1.56 mg/kg, and in corn leaf organ was 2.38 mg/kg.
The results of one-way analysis of variance are presented
in Table 2. According to these results, there is a significant
difference between the measured amounts of nickel,
cadmium, and lead in soil and corn plant organs (root,
stem, and leaf) (P < 0.05). Also, a significant difference
was observed between the amounts of cadmium in
concentrations of 0, 50, and 100 mg/kg with 99%
confidence (P < 0.05).
Table 1. The results of measuring the samples
Samples Statistics
Pb (mg/kg) Cd (mg/kg) Ni (mg/kg)
100 50 0 100 50 0 100 50 0
Soil Samples
Mean 1.56 1.49 1.37 0.44 0.36 0.29 1.51 1.22 1.09
SD 0.042 0.04 0.061 0.055 0.026 0.023 0.1345 0.085 0.11
Corn root samples
Mean 1.72 1.42 0.021 0.58 0.46 0.16 5.79 5.39 2.48
SD 0.037 0.04 0.021 0.083 0.042 0.096 0.3711 0.315 0.059
Corn stalk samples
Mean 1.24 1.03 0.94 0.46 0.39 0.1 > 4.98 4.26 2.08
SD 0.072 0.03 0.011 0.061 0.082 0 0.1124 0.286 0.15
Figure 1. Comparison of measured amounts of nickel in samples (left side: soil samples - right side: corn plant samples)
Figure 2. Comparison of measured lead values in samples (left side: soil samples - right side: corn plant samples)
Arch Hyg Sci. Volume 12, Number 3, 2023 151
Effect of EDTA on heavy metals absorbtion in soil by Corn plants
3.1. Calculate absorption factor
The results of the calculation of TF, BCF, and BAC indices
for the analysis of the absorption of heavy elements by the
organs of the corn plant in different concentrations are
presented in Table 3. In studies to estimate the efficiency
of plants in absorbing heavy elements, the proportion of
the element in the soil to its proportion in the plant organ
is important.
The results of the analysis of nickel element changes
showed that with the increase of soil pollution load to the
level of 50 mg/kg, the absorption level of heavy elements
by leaf and root organs increased. The average BCF index
was 4.44, and BAC was 3.499 (Figure 4). But with the
increase of pollution level to 100 mg/kg of nickel, the
absorption level of heavy elements by these corn plant
organs has decreased. The same situation has happened
in the element cadmium. But in the lead element, with
the increase of pollution concentration to 100 mg/kg, the
absorption level of heavy metals by corn organs has also
increased in a limited way. The most accumulation of
lead was done by the leaf organ. To check the correlation
between heavy metal concentration and indicators,
Pearson’s correlation test was used (Table 4). Its results
showed that there is a significant correlation between the
Figure 3. Comparison of measured amounts of cadmium in samples (left side: soil samples - right side: corn plant samples)
Table 2. The results of the one-way analysis of variance for the studied
variables
Variable Mean square F P value
Nickel (between concentrations) 5.406 1.776 0.173
Nickel (between soil and plant organs) 8.567 2.974 0.033*
Cadmium (between concentrations) 9.097 3.262 0.002*
Cadmium (between soil and plant organs) 6.032 2.046 0.04*
Lead (between concentrations) 5.85 1.876 0.43
Lead (between soil and plant organs) 9.21 2.31 0.01*
* Significance Relation
Figure 4. Comparison of absorption of heavy elements based on TF, BCF, and BAC indices in different concentrations of heavy element pollution (A:
Cadmium-B: Nickel-C: Lead)
Asnaashari et al
152 Arch Hyg Sci. Volume 12, Number 3, 2023
concentration of heavy metals in the organs of the corn
plant with BCF and BAC indices at the level of 0.01.
These results show that, in general, the highest level of
biological accumulation of heavy elements nickel, lead,
and cadmium was related to the organ of the corn leaf.
Also, considering that the average value of the indicators
is more than 1, it can be concluded that corn is a favorable
accumulator of nickel, cadmium, and lead elements, but
the highest level of biological accumulation is related to
nickel elements.
4. Discussion
The use of plants is one of the most effective biological
methods to deal with pollution, such as heavy elements
[38]. One of the consequences of human activities in the
environment is soil pollution, which has also attracted the
attention of natural science researchers [39]. The presence
of heavy elements in the soil (due to their biological
accumulation) is considered a serious threat to the
health of humans and other animals. Therefore, it seems
necessary to use methods to reduce the level of heavy
elements in the soil. Research has confirmed the optimal
efficiency of biological methods, such as bioremediation,
to deal with heavy element pollution [40,41]. The use
of resistant and widely used species in agriculture and
industry can be an effective solution for absorbing soil
pollutants, including heavy elements. According to the
morphological characteristics, such as very long roots
(about 2 m), the corn species can be a good option for
removing heavy elements, which is investigated in the
current research.
The results of other studies, such as Saifullah et al,
Jelusic et al, and Bloem et al, have shown that EDTA
compounds with metals can increase the solubility and
availability of metals in soils [42-44]. When EDTA is used
in soils without creating a strong acidic culture medium,
most of the metals are dissolved and available for green
extraction [45]. The results showed that the most lead
absorption was related to the leaf organ in corn species.
The results of the study by Nazir et al and Shahid et al
also showed that the most lead absorption was done by
the leaves of the orchid and Gluconobacter potus species
[46,47]. In another research, Kumar et al stated that due to
the terrestrial origin of cadmium and nickel elements, the
amounts of these metals in the roots of trees in black spur
forests were more than in other organs [48]. The results
of the present research showed that there is a significant
difference between the measured amounts of nickel at
a concentration of 0 mg/kg with 50 and 100 mg/kg of
soil samples with roots, stems, and leaves of corn plants
(P ≤ 0.05). As a result, under the influence of 2% EDTA,
the bioaccumulation of nickel by the organs of the corn
plant has been at a favorable level. There is a significant
difference between the measured amounts of cadmium
at a concentration of 0 mg/kg with a concentration of
100 mg/kg in the soil and root and stem samples of corn
plants (P ≤ 0.05). But this difference with corn plant leaves
was not significant (P ≥ 0.05). These results show that
EDTA had a favorable effect on the bioaccumulation of
cadmium in corn plant stems and roots at a concentration
of 100 mg/kg.
5. Conclusion
The results of this research and its comparison with other
research show that EDTA can be used to increase the
efficiency of corn plants in absorbing heavy elements of
nickel and cadmium. Also, the absorption of nickel and
cadmium by the roots and lead by the leaves of corn
species has been higher. Due to the existence of many
industrial areas in the south and southwest of Iran, this
method can be used for bioremediation.
Acknowledgments
This article is extracted from the doctoral dissertation in
environmental sciences at the Islamic Azad University of Ahvaz.
We express our sincere gratitude to the officials of the Faculty
of Agriculture and Natural Resources who helped us carry out
and improve the quality of this research. The research code is:
1064817545244271398176458.
Authors’ Contribution
Conceptualization: Kamran Mohseni Far.
Data curation: Mojtaba Alavifazel.
Formal analysis: Mojtaba Alavifazel.
Investigation: Ali Asnaashari.
Methodology: Sima Sabzalipour.
Resources: Sima Sabzalipour.
Software: Mojtaba Alavifazel.
Supervision: Ali Asnaashari.
Validation: Kamran Mohseni Far.
Visualization: Kamran Mohseni Far.
Writing–original draft: Ali Asnaashari.
Writing–review & editing: Kamran Mohseni Far.
Table 3. The results of calculating TF, BAC, and BCF indices
Index Nickel 0 Nickel 50 Nickel 100 Cadmium 0 Cadmium 50 Cadmium 100 Lead 0 Lead 50 Lead 100
TF (index) 0.838 0.793 1.66 - 0.829 0.785 1.361 1.378 1.387
BCF (index) 2.293 4.44 3.85 0.53226 1.295 1.312 1.401 1.483 1.525
BAC (index) 1.926 3.499 3.322 - 1.08 1.029 0.9343 0.953 1.102
Table 4. The results of Pearson correlation analysis between the average
concentration of heavy metals and TF, BCF, and BAC indices
TF BCF BAC
The average
concentration of
heavy metals
The correlation coefficient 0.515 0.948 ** 0.937**
meaningful 0.156 0.000 0.000
** Significance Correlation
Arch Hyg Sci. Volume 12, Number 3, 2023 153
Effect of EDTA on heavy metals absorbtion in soil by Corn plants
Competing Interests
The authors declare that there is no conflict of interest regarding the
publication of this manuscript. Furthermore, the ethical issues have
been completely observed by the authors including plagiarism,
informed consent, misconduct, data fabrication and/or falsification,
double publication and/or submission, and redundancy.
Funding
The present research did not receive any financial support.
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