International Journal of Phytoremediation

ISSN: (Print) (Online) Journal homepage: https://www.tandfonline.com/loi/bijp20

Role of magnesium oxide nanoparticles in the

mitigation of lead-induced stress in Daucus
carota: modulation in polyamines and antioxidant
enzymes

Samia Faiz, Nasim Ahmad Yasin, Waheed Ullah Khan, Anis Ali Shah, Waheed
Akram, Aqeel Ahmad, Aamir Ali, Naima Huma Naveed & Luqman Riaz

To cite this article: Samia Faiz, Nasim Ahmad Yasin, Waheed Ullah Khan, Anis Ali Shah, Waheed
Akram, Aqeel Ahmad, Aamir Ali, Naima Huma Naveed & Luqman Riaz (2022) Role of magnesium
oxide nanoparticles in the mitigation of lead-induced stress in Daucus�carota: modulation in
polyamines and antioxidant enzymes, International Journal of Phytoremediation, 24:4, 364-372,
DOI: 10.1080/15226514.2021.1949263

To link to this article: https://doi.org/10.1080/15226514.2021.1949263

Published online: 20 Jul 2021. Submit your article to this journal

Article views: 126 View related articles

View Crossmark data Citing articles: 3 View citing articles

Full Terms & Conditions of access and use can be found at

https://www.tandfonline.com/action/journalInformation?journalCode=bijp20
INTERNATIONAL JOURNAL OF PHYTOREMEDIATION
2022, VOL. 24, NO. 4, 364–372
https://doi.org/10.1080/15226514.2021.1949263

Role of magnesium oxide nanoparticles in the mitigation of lead-induced stress

in Daucus carota: modulation in polyamines and antioxidant enzymes
Samia Faiza, Nasim Ahmad Yasinb , Waheed Ullah Khanc, Anis Ali Shahd, Waheed Akrame , Aqeel Ahmade,
Aamir Alia, Naima Huma Naveeda, and Luqman Riazf
a
Department of Botany, University of Sargodha, Sargodha, Pakistan; bSenior Superintendent Garden, RO-II Office, University of the Punjab,
Lahore, Pakistan; cDepartment of Environmental Sciences, Islamia University, Bahawalpur, Pakistan; dDepartment of Botany, University of
Narowal, Narowal, Pakistan; eGuangdong Key Laboratory for New Technology Research of Vegetables, Vegetable Research Institute,
Guangdong Academy of Agricultural Sciences, Guangzhou, China; fCollege of Life Sciences, Henan Normal University, Xinxiang, China

ABSTRACT KEYWORDS
During the current study, the effects of magnesium oxide nanoparticles (5 mmol/L) were observed Antioxidant; carrot;
on the growth and mineral nutrients of Daucus carota under lead (Pb) stress. The results demon- nanoparticles; stress
strated that Pb stress decreased the growth and photosynthetic rate of D. carota plants.
Furthermore, Pb stressed plants showed decreased uptake of mineral nutrients including Zn, Na, Fe,
K, Ca, Mg, K, and Cu. Similarly, Pb stressed plants showed enhanced electrolyte leakage (EL) and
malondialdehyde (MDA) content. However, magnesium oxide nanoparticles detoxified ROS to miti-
gate Pb stress and improved the growth of plants. Magnesium oxide nanoparticles also escalated the
activity of antioxidant enzymes including superoxide dismutase (SOD) and Catalase (CAT). A higher
amount of Pb content was observed in the roots as compared to the shoot of plants. Lead toxicity
reduced manganese accumulation in D. carota plants. The increased concentration of iron, manga-
nese, copper, and zinc advocates stress the ameliorative role of Pb stress in plants.

Novelty statement
The role of MgONPs in the alleviation of Pb-toxicity in Daucus carota has never been exploited. In
addition, the potential of MgONPs to enhance nutritional content in D. carota via modulation in
antioxidant system and polyamines have never been reported.

Introduction carbohydrate formation in plants (Hermans et al. 2004;

At present, heavy metal contamination is a major challenge
for the development of sustainable programs. Lead (Pb) is
considered the second most injurious metal for all living
organisms. Pb-contamination due to automobile exhaust
besides the use of Pb-contaminated fertilizers has become a
protuberant issue throughout the world. The unremitting
application of industrial effluents polluted drain water for
irrigation leads to Pb contamination in agricultural sites.
Most of the plants exhibit toxicity symptoms when Pb con-
centration exceeds 30 mg/kg soil (Yasseen and Al-Thani
2013). Furthermore, the consumption of crops growing on
metal-contaminated sites causes metal toxicity in plants and
animals (Khan et al. 2019). Metal stress restricts the uptake
and translocation of mineral cations including copper (Cu),
manganese (Mn), zinc (Zn), and iron (Fe) (Ferrol Gonz
alez
et al. 2016). Whereas, higher bioavailability of these plant
nutrients will decrease the uptake of metal contaminants
(Choppala et al. 2014).
Magnesium (Mg) plays a vital role in the normal growth
of plants and animals. The deficiency of this essential nutri-
ent negatively affects photosynthetic activity and
Nejia et al. 2016). About 35% of foliage Mg is present in
leaf chloroplast (Cakmak and Yazici 2010). The grana devel-
opment in the chloroplast is not possible in absence of Mg.
Correspondingly, this nutrient exerts a negative surface
charge on the thylakoid membranes resulting in Mg deposits
(Puthiyaveetil et al. 2017). It is also a prerequisite for
chlorophyll synthesis (Masuda 2008). Equally, Mg activates
Rubisco through establishing a complex with carbamate
group of Rubisco and Rubisco activase (Portis 2003). Plants
deficient in Mg exhibit chlorotic symptoms and yield loss
due to reduction in transport of carbohydrates from source
to sink tissues (Cakmak and Yazici 2010; Gransee and F€ uhrs
2013). Magnesium fertilizers are applied for the growth and
yield enhancement of Mg deficient plants (Nedim and Daml
2015). Extensive study material is accessible describing the
significance of this vital element in physiochemical progres-
sions of plants (Cakmak 2013; Ceylan et al. 2016).
Nevertheless, it is essential to recognize the potential of Mg
nanoparticles on the growth and metal stress alleviation of
carrots. Up till now, no research work has been carried out
to thoroughly investigate the influences of Mg nanoparticles
on plant growth under Pb stress. The current study was

CONTACT Nasim Ahmad Yasin nasimhort@gmail.com Senior Superintendent Garden, RO-II Office, University of the Punjab, Lahore, Pakistan.
ß 2021 Taylor & Francis Group, LLC

designed to observe the potential of Mg nanoparticles in the
improvement of growth and Pb stress mitigation in carrots.
Carrot (Daucus carota subsp. sativus L.) which belongs to
Apiaceae family is one of the commonly cultivated vegeta-
bles of the world. The yield of carrots estimated during 2017
was 42.8 million tons (FAO Stats 2017). Carrot is necessary
for eyesight because it contains carotenoids which are pre-
cursors of vitamin A (Kopsell and Kopsell 2006). It also
contains vitamins B, C, K besides protein and dietary fibers
(Arscott and Tanumihardjo 2010; Sharma et al. 2012).
Carrot is used to prepare salads, soups, and culinary (Atkins
1999; Mateljan 2007). Carrot requires relatively low fertilizer
requirements; hence farmers having low financial resources
prefer to cultivate this valuable crop (Allemann and Young
2002). The growth, yield, and quality of carrot cultivars are
dependent on fertilization and environmental characteristics
(Singh et al. 2012). The progressive farmers apply enormous
fertilizers to obtain appropriate produce of carrot.
Conversely, these synthetic fertilizers are causing health and
environmental issues (Arisha and Bradisi 1999; Agbede et al.
2017). Furthermore, several crops growing in Pb polluted
soil regimes exhibit decreased mineral nutrition, growth,
and quality. Recently, the applications of nanoparticles have
exhibited gigantic potential in agriculture. Yet, there is no
study describing the influence of Mg nanoparticles on
nutrients homeostasis in carrots subjected to Pb stress. It is
essential to understand the capability of Mg nanoparticles to
assuage Pb stress in crops. Hence, the carrot was used as a
model crop to discover the Pb stress alleviating role of Mg
nanoparticles. Furthermore, the role of Mg nanoparticles on
uptake and accumulation of Pb as well as plant nutrients
was also studied during the current study.
Materials and methods
Plant materials and growth conditions
The present research work was carried out in the
Department of Botany, University of Sargodha. Soil samples
were analyzed using stainless steel auger from 0 to 20 cm
depth. Soil samples contain organic content 2.87 g kg
1,
potassium (K) 2.38 g kg
1, nitrogen (N) 1.23 g kg
1, 0.03 g
kg
1 Pb. For Pb contamination, 500 mg kg
1 Pb (NO3)2 salt
was mixed in the soil of each pot and placed in the shade
for 15 days to attain equilibrium. Magnesium oxide nanopar-
ticles (5 mmol/L) were purchased from Sigma Aldrich.
Magnesium oxide nanoparticles are distributed in deionized
water and poured into potted water. Plants were harvested
after 40 days and growth parameters were analyzed.
Estimation of photosynthetic rate
The amount of net rate of photosynthesis (Pn) was analyzed
at 9 am from the second upper fully opened leaf by using
an infrared gas analyzer (LiCor, Lincoln, NE, USA). The
relative humidity during the observation was 65 ± 4%,
photosynthetically active radiation was adjusted at
INTERNATIONAL JOURNAL OF PHYTOREMEDIATION 365
1,000 ± 6 lmol m
2 s
1, the temperature was 25 ± 3  C and
atmospheric CO2 was kept at 355 lmol mol
1.
Estimation of electrolyte leakage
The prewashed leaves were divided into 0.5 cm2 segments.
These leaf segments were placed in flasks containing 10 mL
of demineralized water and electrolyte leakage of this solu-
tion was observed after 22 h at 25  C with the help of a con-
ductometer. Afterward, flasks were placed in the oven at
90  C for 120 min to acquire total conductivity.
Estimation of hydrogen peroxide
Foliage samples (100 mg) were mixed in 0.4 g activated char-
coal along with 12 mL of 5% TCA in an ice-chilled mortar.
The mixture was centrifuged at 13,000 rpm for 15 min at 4  C.
The 0.5 mL supernatant was dissolved in 1 mL of 1 M potas-
sium iodide in presence of 0.5 mL of 5 mM KH2PO4 buffer at
pH 7. The amount of H2O2 was measured by taking absorb-
ance at 390 nm.
Evaluation of malondialdehyde level and
antioxidative activities
The 1 g freshly harvested plant sample was vortexed with
20 mL of sodium phosphate buffer at pH 7.8 by using an ice-
cold mortar. This solution was centrifuged for 10 min at
10,000  g. The upper layer was separated and used for the
estimation of peroxidase activity (POD) and catalase activity
(CAT). For assessment of POD activity, the absorbance of
6 mL of a reaction mixture comprising enzyme extract, H2O2,
PBS, and guaiacol was estimated at 470 nm. For estimation of
CAT activity, the absorbance of reaction solution containing
enzyme extract, deionized water, and H2O2 and was analyzed
at 240 nm.
The MDA level was estimated according to Heath and
Packer (1968). For this purpose, 1 g plant sample was homo-
genized with 5% trichloroacetic acid (TCA) and subjected to
15 min centrifugation at 10,000  g. The reaction mixture
included plant extract, PBS, and TBA solution. The solution
was placed at 98  C for 20 min, followed by centrifugation,
and then MDA level was observed at 600, 532, and 450 nm.
Estimation of PAs
The PAs were quantified by performing Liquid
Chromatography-Mass Spectrometry (LC-MS) with the help
of a Shimadzu HP series 31. The PAs were separated by using
C18 Column (Phenomenox). The gradient developed among
acetonitrile and water, acidified by using 0.l% formic acid was
employed as a mobile phase to separate compounds.
Estimation of soluble sugar content
Plant samples (5 gm) were shaken filtered with the help of
filter paper. Sugar content was estimated using a
refractometer.
366 S. FAIZ ET AL.
Analysis of terpenoid
Root samples were obtained for the measurement of terpen-
oid content. Plant samples (50 g) were blended using water
for 2 min, 1 mL of headspace gas was injected into a gas
chromatograph (PerkinElmer 8700, Norwalk, CT, USA)
equipped with a flame ionization detector. Monoterpenoid
content of myrcene, b-pinene, a-pinene, c-terpenine, limon-
ene, terpinolene were measured and added to quantify total
terpenoid content. Following injection of gas samples, slurry
samples (2 and 5 g) were collected and placed into the plas-
tic tube. Afterward, the obtained samples were kept at

20  C for analysis. Plant samples were deforested and
homogenized with extraction solvents using a homogenizer
(PT 10-35, Brinkmann, Westbury, NY, USA).
Analysis of carotenoid content
Quantification of carotenoid content was carried out as per
the method described by Yoo et al. (2012). Hexane extracts
of root and plant samples were obtained with the help of
steamed nitrogen, added 0.5 mL acetone, and injected into
HPLC (PerkinElmer 200 series) system. A spherisorb-ODS2
column (Alltech, Deerfeld, IL, USA) and a mobile phase of
ethyl acetate (35%) in water-acetonitrile containing 0.1%
triethylamine) were used at the rate of 1 mL min
1.
Detection of carotenoid content was carried out at 450 nm.
The spectral data of each carotenoid was measured between
200 and 700 nm and compared with standard spectra of the
compound (Britton 1991). For quantification of carotenoid
content, external standards from Sigma-Aldrich (St. Louis,
MO, USA) were utilized.
Analysis of anthocyanin content
Plant samples (50 g) were blended with 50 mL water with
HCL (1%) for 2.5 min. 5 g slurry samples were frozen at

20  C. Plant samples were deforested and homogenized
with HCL (100 mL 1% HCL) in water and transferred to a
cylinder, making total volume up to 100 mL. The homogen-
ous mixture was filtered through a 0.45 mm filter and the
absorbance value was determined at 530 nm to calculate
total anthocyanin content using an extinction coefficient. In
case of absorbance value over 1, the plant sample was
diluted with HCL (1%).
Table 1. Effect of magnesium oxide nanoparticles on growth attributes of D. carota under Pb-stressed soil.
Treatments Leave fresh weight (g) Root fresh weight (g)
C 15.64 ± 0.72b 17.25 ± 0.92ab
Pb 8.92 ± 0.51bc 12.83 ± 0.73b
MgONPs 19.34 ± 1.16a 20.26 ± 0.96a
Pb þ MgONPs 10.27 ± 0.52c 14.62 ± 0.83b
C: control; Pb: lead; MgONPs: 5 mmol/L magnesium oxide nanoparticles.
Values are means ± SD of five replicates. Different letters in the same column show significant differences among treatments at p  0.05.
Minerals and lead analysis
Plant samples were oven-dried and then digested using
H2SO4/HNO3 (1/5 v/v) for 24 h followed by mixing with
HNO3/HCLO4 mixture (5/1, v/v). Mineral nutrients were
quantified using atomic absorption spectrophotometer. Lead
content in digested samples was estimated with the help of
atomic absorption spectrophotometer.
Statistical analysis
The data comprising of average values of 5 replicates were
subjected to one-way ANOVA and consequently to
Duncan’s Multiple Range Test. The variations were regarded
as significant when the p-value was at least 0.05. Lower
case letters were used to emphasize significant differences
among the treatments using DSASTAT software.
Results
Influence of MgO nanoparticles on growth attributes of
D. carota under Pb stress
During the present study, lead stress exerted devastating
effects on the growth of D. carota plants. The Pb toxicity
reduced the root dry weight, leaves dry weight, root length,
and shoot length by 31, 43, 23, and 22%, respectively as
compared to control. However, magnesium oxide nanopar-
ticles application improved the root dry weight, leaves dry
weight, root length, and shoot length of D. carota by 21, 19,
15, and 18%, respectively under metal stressed regimes for
only Pb treated plants (Table 1).
Effect of MgO nanoparticles on the levels of chlorophyll,
anthocyanins, soluble sugar content (SSC), terpenoid,
and carotene content of D. carota
The results of the current study showed that Pb toxicity
remarkably modulated the levels of chlorophyll contents,
anthocyanins, soluble sugar content, terpenoid, and carotene
values in D. carota plants to control (Table 2). Nevertheless,
MgO nanoparticles supplementation enhanced the levels of
these biochemicals in D. carota plants under toxic and non-
toxic conditions. The Pb þ MgONPs treatment increased the
levels of chlorophyll, anthocyanins, soluble sugar content,
terpenoid, a-carotene, and b-carotene contents in D. carota
plants by 13, 15, 9, 11, 12, and 16%, respectively, when com-
pared with only Pb subjected plants.
Growth parameters
Leave dry weight (g) Root dry weight (%) Root length (cm) Shoot length (cm)
4.87 ± 0.28ab 6.75 ± 0.45ab 10.28 ± 0.72ab 14.27 ± 0.74b
2.78 ± 0.17c 4.63 ± 0.34b 7.96 ± 0.58c 11.18 ± 0.58c
5.68 ± 0.32a 7.28 ± 0.42a 12.53 ± 0.82a 17.83 ± 0.92a
3.42 ± 0.18b 5.86 ± 0.62b 9.27 ± 0.61b 13.49 ± 0.72c
 INTERNATIONAL JOURNAL OF PHYTOREMEDIATION 367

Table 2. Effect of magnesium oxide nanoparticles on the levels of chlorophyll, anthocyanins, soluble sugar content (SSC), terpenoid, and carotene content of D.
carota under Pb-stressed soil.
Chlorophyll Anthocyanins SSC Terpenoid a-carotene (leaves) a-carotene (root) b-carotene (leaves) b-carotene (root)
Treatments mg g
1 lg g
1 FW (mg
1 DW) mg g
1 FW mg g
1 FW mg g
1 FW mg g
1 FW mg g
1 FW
C 16.52 ± 0.92b 4.69 ± 0.31ab 5.8 ± 0.29ab 14.38 ± 0.79ab 1.24 ± 0.065ab 4.18 ± 0.53ab 25.16 ± 1.29b 28.27 ± 1.79b
Pb 11.74 ± 0.63c 3.57 ± 0.18b 4.52 ± 0.25b 11.24 ± 0.62b 1.05 ± 0.052c 3.92 ± 0.28b 19.38 ± 1.08c 22.87 ± 1.16c
MgONPs 21.68 ± 1.24a 5.24 ± 0.27a 6.79 ± 0.34a 15.85 ± 0.82a 1.37 ± 0.071a 4.36 ± 0.31a 29.84 ± 1.48a 33.62 ± 2.34a
Pb þ MgONPs 13.29 ± 0.74c 4.19 ± 0.26b 4.98 ± 0.27b 12.56 ± 0.68b 1.18 ± 0.062b 3.98 ± 0.19b 23.29 ± 1.17bc 25.41 ± 1.42bc
C: control; Pb: lead; MgONPs: 5 mmol/L magnesium oxide nanoparticles.
Values are means ± SD of five replicates. Different letters in the same column show significant differences among treatments at p  0.05.

a
Net Photosynthetic rate (µ mol CO 2 m-2 S-1)

4 b

bc
3

2 c

0
C Pb MgONPs Pb + MgONPs

Figure 1. Influence of magnesium oxide nanoparticles on the photosynthetic rate of D. carota under Pb-stressed soil. C: Control; Pb: Lead; MgONPs: 5 mmol/L mag-
nesium oxide nanoparticles. Values are means ± SD of five replicates. Different letters in the same column show significant differences among treatments
at p  0.05.

Impact of MgO nanoparticles on photosynthetic rate of MDA, and H2O2, respectively, in D. carota plants subjected
D. carota plants to Pb stress in comparison with analogous controls.
Lead stress declined the net photosynthetic rate in D. carota
plants by 53% than that of untreated control. However, Implications of MgO nanoparticle on antioxidative
MgO nanoparticle supplementation augmented the net enzyme activities in D. carota plants
photosynthetic rate in D. carota plants during toxic and
The findings of the present study indicated that Pb toxicity
non-toxic conditions. The MgO nanoparticles treatment
significantly augmented the activities of antioxidant
increased the net photosynthetic rate in D. carota plants
enzymes, such as superoxide dismutase (SOD) and catalase
raised under the Pb regime by 38% to relevant Pb control (CAT) in D. carota plants by 29 and 32%, respectively when
(Figure 1). compared with corresponding untreated controls. Whereas,
MgONPs supplementation further improved the activities of
SOD and CAT in D. carota plants during noxious and non-
Influence of MgO nanoparticles on electrolyte leakage noxious situations (Figure 3). The Pb þ MgONPs treatment-
(EL), hydrogen peroxide (H2O2), and malondialdehyde induced 29 and 25% increment in the activities of SOD and
CAT in D. carota plants exposed to Pb stress in contrast
(MDA) of D. carota plants
with corresponding metal controls.
During the current investigation, D. carota plants grown in
Pb toxic regimes exhibited enhancement in the level of EL,
Effect of MgO nanoparticle over polyamines (PAs) in D.
MDA and H2O2 by 49, 28, and 43%, respectively, compared
carota plants under Pb stress
with untreated control. Nevertheless, MgO nanoparticles
treated plants demonstrated remarkable diminishment in the The current results depicted that Pb toxicity boosted the lev-
values of EL, H2O2, and MDA during stressed and non- els of polyamines in D. carota plants as compared to
stressed circumstances (Figure 2). The Pb þ MgONPs treat- untreated control. However, MgO nanoparticle supplemen-
ment showed 22, 31, and 19% reduction in the levels of EL, tation induced further enhancement in the values of
368 S. FAIZ ET AL.

60
a

Electrolyte leakage
50
b
40

30 c

20
d
10

8
a
7

6
MDA (mM g-1 FW)

b
5
bc
4
c
3

50 a
45
40
b
H2O2 (µ MOL g-1 FW)

35
30
bc
25
20
c
15
10
5
0
C Pb MgONPs Pb + MgONPs
Treatments
Figure 2. Influence of magnesium oxide nanoparticles on EL, MDA, and H2O2 content of D. carota under Pb-stressed soil. C: Control; Pb: Lead; MgONPs: 5 mmol/L
magnesium oxide nanoparticles. Values are means ± SD of five replicates. Different letters in the same column show significant differences among treatments
at p  0.05.

polyamines in D. carota plants in case of contaminated and
non-contaminated regimes. The Pb þ MgONPs treatment
revealed a 16% higher value of polyamines in D. carota
plants over the corresponding only metal amended one
(Figure 4).
regimes (Tables 3–4). The Pb þ MgONPs treatment resulted
in 16, 11, 8, 15, 12, 10, 21, 9, and 14% increment in the level
of minerals; Zn, Cu, K, P, Ca, Mg, S, N, and Mn in the
roots of D. carota plants, respectively than that of only Pb
exposed ones.

Effect of magnesium oxide nanoparticles on mineral
uptake of D. carota under Pb-stress
During the present investigation, it was noted that Pb tox-
icity interfered with the mineral uptake capability of root
and above-ground parts of D. carota plants. The Pb stressed
D. carota plants exhibited noticeably reduced uptake of Zn,
Cu, K, Ca, Mg, S, and Mn in root and leave tissues as com-
pared to control. However, MgO nanoparticles supple-
mented D. carota plants demonstrated perceptible
enhancement in the uptake of Zn, Cu, K, P, Ca, Mg, S, N,
and Mn in root and leave tissues during toxic and non-toxic
Effect of magnesium oxide nanoparticles on Pb uptake
in root and leaves/shoot, translocation factor (TF), bio-
concentration factor (BCF), and tolerance index (TI) of
D. carota under Pb-stress
The root tissues of D. carota plants showed relatively higher
Pb accumulation as compared to leaves/shoot tissues. The
MgO nanoparticles supplementation lowered Pb uptake in
root and above-ground parts of D. carota plants. The MgO
nanoparticles treated D. carota plants grown under Pb stress
exhibited reduced values of TF and BCF as compared to
only Pb subjected ones (Table 5). However, MgO
 INTERNATIONAL JOURNAL OF PHYTOREMEDIATION 369

0.4
a
0.35

SOD activity (U/ mg protein)

0.3
bc
0.25
c
0.2

0.15

0.1

0.05

7 a

6
CAT activity (µ mol min-1 mg-1 protein )

b
5
bc

4
c

0
C Pb MgONPs Pb + MgONPs

Treatments

Figure 3. Influence of magnesium oxide nanoparticles on SOD and CAT activity of D. carota under Pb-stressed soil. C: Control; Pb: Lead; MgONPs: 5 mmol/L magne-
sium oxide nanoparticles. Values are means ± SD of five replicates. Different letters in the same column show significant differences among treatments at p  0.05.

1200

a
Total polyamine content Fluor. Intensity (a.u)

1000 b

800
bc
c
600

400

200

0
C Pb MgONPs Pb + MgONPs

Treatments
Figure 4. Influence of magnesium oxide nanoparticles on total polyamine content of D. carota under Pb-stressed soil. C: Control; Pb: Lead; MgONPs: 5 mmol/L mag-
nesium oxide nanoparticles. Values are means ± SD of five replicates. Different letters in the same column show significant differences among treatments
at p  0.05.
370 S. FAIZ ET AL.

Table 3. Effect of magnesium oxide nanoparticles on mineral uptake in the root of D. carota under Pb-stressed soil.
Root Mineral uptake (mmol kg
1 DW)
Treatments Zn Cu K P Ca Mg S N Mn
C 82 ± 6b 5.36 ± 0.29b 310 ± 15b 65 ± 4c 75 ± 5b 37 ± 2.5ab 19 ± 1.54b 590 ± 27c 39 ± 2.84b
Pb 56 ± 5c 4.43 ± 0.28c 265 ± 13d 82 ± 6ab 64 ± 4c 29 ± 1.9c 15 ± 0.92c 635 ± 31b 28 ± 1.93d
MgONPs 92 ± 7a 6.72 ± 0.38a 353 ± 16a 78 ± 5b 86 ± 6a 45 ± 3.2a 26 ± 1.23a 628 ± 29bc 46 ± 3.27a
Pb þ MgONPs 67 ± 5bc 4.96 ± 0.31bc 294 ± 14c 96 ± 6a 72 ± 5bc 32 ± 2.6bc 19 ± 0.87bc 689 ± 28a 32 ± 1.85c
C: control; Pb: lead; MgONPs: 5 mmol/L magnesium oxide nanoparticles.
Values are means ± SD of five replicates. Different letters in the same column show significant differences among treatments at p  0.05.

Table 4. Effect of magnesium oxide nanoparticles on mineral uptake in leaves of D. carota under Pb-stressed soil.
Leaves Mineral uptake (mmol kg
1 DW)
Treatments Zn Cu K P Ca Mg S N Mn
C 105 ± 6.2b 7.24 ± 0.37b 478 ± 23bc 42 ± 2.54d 375 ± 18b 92 ± 4.58b 52 ± 3.26b 590 ± 31d 62 ± 4.12b
Pb 87 ± 4.5d 6.18 ± 0.38c 462 ± 21c 63 ± 3.72b 332 ± 16c 78 ± 3.92c 31 ± 2.14c 635 ± 34c 48 ± 3.38d
MgONPs 121 ± 7.3a 8.31 ± 0.45a 506 ± 26a 54 ± 3.25c 415 ± 21a 109 ± 5.23a 61 ± 3.48a 657 ± 29b 71 ± 5.21a
Pb þ MgONPs 94 ± 5.1c 6.95 ± 0.41bc 483 ± 24b 71 ± 4.31a 362 ± 19bc 85 ± 4.17bc 36 ± 2.06bc 672 ± 31a 54 ± 3.28c
C: control; Pb: lead; MgONPs: 5 mmol/L magnesium oxide nanoparticles.
Values are means ± SD of five replicates. Different letters in the same column show significant differences among treatments at p  0.05.

Table 5. Effect of magnesium oxide nanoparticles on lead (Pb) uptake in root and leaves/shoot, translocation factor (TF), bio-concentration
factor (BCF) and tolerance index (TI) of D. carota under Pb-stressed soil.
Treatments Root Pb uptake Leaves þ shoot Pb uptake TF BCF TI (%)
C ND ND – – –
Pb 9.92 ± 0.51a 6.83 ± 0.31a 0.68 ± 0.027a 0.27 ± 0.013a 78 ± 5.86 cc
MgONPs 0.34 ± 0.016c 0.23 ± 0.01c 0.67 ± 0.028a 0.15 ± 0.01b 123 ± 8.52a
Pb þ MgONPs 7.28 ± 0.42b 4.32 ± 0.83b 0.59 ± 0.026b 0.172 ± 0.01ab 92 ± 6.74b
C: control; Pb: lead; MgONPs: 5 mmol/L magnesium oxide nanoparticles.
Values are means ± SD of five replicates. Different letters in the same column show significant differences among treatments at p  0.05.

nanoparticles enhanced TI. A Higher TI value was observed
in MgO nanoparticles applied to plants under stress as com-
pared to those subjected to metal toxicity.
Discussion
Roots are the first points of plants that uptake Pb and
exhibit a higher concentration of this metal compared to
foliage parts of plants (Wierzbicka et al. 2007). Lead toxicity
reduces photosynthetic pigments and uptake of nutrients
including Zn, Ca, Mn, and Fe (Eun et al. 2000; Rout and
Das 2003). Furthermore, Pb-stressed plants show a reduced
level of Mg content (H€aussling et al. 1998). Kosobrukhov
et al. (2004) observed a decline in biomass production in
plants affected by Pb toxicity. While Akinci et al. (2010)
reported a reduction in root and shoot growth of tomato
plants grown under Pb contaminated conditions. Poor nutri-
tion and reduced photosynthesis result in the decrease of
growth and biomass in Pb stressed plants (Lamhamdi et al.
2011). During the current study, Pb toxicity reduced growth
and photosynthetic rate in D. carota plants.
Lead toxicity impedes the uptake and translocation of
mineral nutrients in affected plants. The current study also
depicted a decrease in nutritional content in D. carota plants
grown under Pb-stressed conditions. The superfluous ionic
leakage from plant tissues resulted in a reduction in miner-
als uptake.
From our results, it is evident that Mg improved the rate
of photosynthesis in D. carota plants. Hermans et al. (2011)
also reported that that Mg alleviated Cd stress by
stabilization of chlorophyll structure. The higher level of
ROS synthesized in stressed plants causes photo-inhibition
by the photosynthetic electron transport chain (Critchley
1981). Yet, the possible defensive role of Mg in reducing
photo-oxidation of the photosynthetic membrane through
detoxification of ROS in Pb stressed plants need further
clarifications. The Mg may have reduced the chloroplast
destruction in stressed plants resulting in improved photo-
synthetic activity. Magnesium improves metal tolerance in
plants by reducing uptake and translocation of injurious
metals, such as Al and Cd (Kashem and Kawai 2007; Bose
et al. 2011). The data obtained during the current study also
reveals that Mg reduced Pb uptake and accumulation in
applied plants. Kashem and Kawai (2007) also verified that
Mg reduced metal contents in treated Brassica rapa plants.
Our results also suggest that Mg enhanced the roots absorp-
tion area which in return improved uptake of essential min-
erals and water, favoring plant growth and development
(Glinka 1980). Additionally, Mg may have improved the cel-
lular integrity to mitigate Pb stress. Hence, the application
of Mg is a promising technique to mitigate Pb stress in car-
rot plants through improving photosynthetic activity, anti-
oxidant activity, and growth of plants.
Lead stress modulated the activity of antioxidative
enzymes in Pisum sativum seedlings (Dias et al. 2019).
Likewise, during current research, Pb toxicity modulated the
activity of antioxidant enzymes. Sharma et al. (2019)
reported that nanoparticles enhance phyto-tolerance to oxi-
dative damage by increasing the activity of antioxidant
enzymes. The current study also showed that MgONPs

enhanced the activity of antioxidant enzymes in D. car-
ota plants.
Anthocyanins are natural pigments that play a pivotal
role in protection against biotic and abiotic stresses (Zhang
et al. 2019). Carotenes also protect plants from numerous
environmental stresses through modulations in membrane
physical properties (Strzałka et al. 2003). This study also
showed that MgONPs enhanced anthocyanins and caroten-
oid content in D. carota plants as compared to con-
trol conditions.
Polyamines are regarded as crucial bio-stimulants which
play an important role in the growth of plants (Chen et al.
2018). The current study showed that MgONPs enhanced
total polyamine content in D. carota plants grown in normal
and Pb-contaminated conditions. Hence, the increased syn-
thesis of polyamines besides modulated activity of antioxi-
dants alleviated Pb stress in MgONPs treated carrot plants.
Conclusions
From the results of the present study, it is concluded that
Pb-induced stress reduced the root and shoot growth of car-
rot plants. Moreover, Pb decreased Fe, Zn, and Cu contents
in stressed plants. However, exogenously applied MgONPs
(5 mmol/L) alleviated Pb stress by reducing the uptake and
translocation of Pb. Additionally, Mg applied plants exhib-
ited higher Fe, Zn, and Cu contents in observed plant tis-
sues. Our results exhibit that increased Pb content disturbs
uptake of plant nutrients which in return impede normal
physiochemical parameters of plants. Moreover, reduced
photosynthetic activity and growth were observed in plants
subjected to Pb stress.
ORCID
Nasim Ahmad Yasin http://orcid.org/0000-0002-1897-0959
Waheed Akram http://orcid.org/0000-0003-3811-6677
References
Agbede TM, Adekiya AO, Eifediyi EK. 2017. Impact of poultry manure
and NPK fertilizer on soil physical properties and growth and yield
of carrot. J Hortic Res. 25(1):81–88. doi:10.1515/johr-2017-0009.
Akinci IE, Akinci S, Yilmaz K. 2010. Response of tomato (Solanum
lycopersicum L.) to lead toxicity: growth, element uptake, chlorophyll
and water content. Afr J Agric Res. 5(6):416–423.
Allemann L, Young BW. 2002. Anintroduction to vegetable production.
Nutrition, fertilisers, organic manures and compost making. 3rd ed.
Pietermaritzburg: Department of Agriculture and Environmental
Affairs.
Arisha HM, Bradisi A. 1999. Effect of mineral fertilizers and organic
fertilizers on growth, yield and quality of potato under sandy soil
conditions. Zagazig J Agric Res. 26:391–405.
Arscott SA, Tanumihardjo SA. 2010. Carrots of many colors provide
basic nutrition and bioavailable phytochemicals acting as a func-
tional food. Compr Rev Food Sci Food Saf. 9(2):223–239. doi:10.
1111/j.1541-4337.2009.00103.x.
Atkins L. 1999. Calcium content of raw vegetables. USA: United States
Department of Agriculture. Agricultural Research Service.
Britton G. 1991. The biosynthesis of carotenoids: a progress report.
Pure Appl Chem. 63(1):101–108. doi:10.1351/pac199163010101.
INTERNATIONAL JOURNAL OF PHYTOREMEDIATION 371
Bose J, Babourina O, Rengel Z. 2011. Role of magnesium in alleviation
of aluminium toxicity in plants. J Exp Bot. 62(7):2251–2264. doi:10.
1093/jxb/erq456.
Cakmak I. 2013. Magnesium in crop production, food quality and
human health. Plant Soil. 368(1–2):1–4. doi:10.1007/s11104-013-
1781-2.
Cakmak I, Yazici AM. 2010. Magnesium: a forgotten element in crop
production. Better Crops. 94(2):23–25.
Ceylan Y, Kutman UB, Mengutay M, Cakmak I. 2016. Magnesium
applications to growth medium and foliage affect the starch distribu-
tion, increase the grain size and improve the seed germination in
wheat. Plant Soil. 406(1–2):145–156. doi:10.1007/s11104-016-2871-8.
Chen D, Shao Q, Yin L, Younis A, Zheng B. 2018. Polyamine function
in plants: metabolism, regulation on development, and roles in abi-
otic stress responses. Front Plant Sci. 9:1945. doi:10.3389/fpls.2018.
01945.
Choppala G, Saifullah Bolan N, Bibi S, Iqbal M, Rengel Z,
Kunhikrishnan A, Ashwath N, Ok YS. 2014. Cellular mechanisms in
higher plants governing tolerance to cadmium toxicity. Crit Rev
Plant Sci. 33(5):374–391.
Critchley C. 1981. Studies on the mechanism of photoinhibition in
higher plants: I. Effects of high light intensity on chloroplast activ-
ities in cucumber adapted to low light. Plant Physiol. 67(6):
1161–1165. doi:10.1104/pp.67.6.1161.
Dias MC, Mariz-Ponte N, Santos C. 2019. Lead induces oxidative stress
in Pisum sativum plants and changes the levels of phytohormones
with antioxidant role. Plant Physiol Biochem. 137:121–129. doi:10.
1016/j.plaphy.2019.02.005.
Eun SO, Shik Youn H, Lee Y. 2000. Lead disturbs microtubule organ-
ization in the root meristem of Zea mays. Physiol Plant. 110(3):
357–365. doi:10.1034/j.1399-3054.2000.1100310.x.
FAO Stats. 2017. http://www.fao.org/.
Ferrol Gonz
alez N, Tamayo E, Vargas Gallego PA. 2016. The heavy
metal paradox in arbuscular mycorrhizas: from mechanisms to bio-
technological applications.
Glinka Z. 1980. Abscisic acid promotes both volume flow and ion
release to the xylem in sunflower roots. Plant Physiol. 65(3):
537–540. doi:10.1104/pp.65.3.537.
Gransee A, F€ uhrs H. 2013. Magnesium mobility in soils as a challenge
for soil and plant analysis, magnesium fertilization and root uptake
under adverse growth conditions. Plant Soil. 368(1–2):5–21. doi:10.
1007/s11104-012-1567-y.
H€aussling M, Jorns CA, Lehmbecker G, Hecht-Buchholz C, Marschner
H. 1988. Ion and water uptake in relation to root development in
Norway spruce (Picea abies (L.) Karst.). J Plant Physiol. 133(4):
486–491. doi:10.1016/S0176-1617(88)80042-7.
Heath RL, Packer L. 1968. Photoperoxidation in isolated chloroplasts.
Arch Biochem Biophys. 125(1):189–198. doi:10.1016/0003-
9861(68)90654-1.
Hermans C, Chen J, Coppens F, Inz
e D, Verbruggen N. 2011. Low
magnesium status in plants enhances tolerance to cadmium expos-
ure. New Phytol. 192(2):428–436. doi:10.1111/j.1469-8137.2011.
03814.x.
Hermans C, Johnson GN, Strasser RJ, Verbruggen N. 2004. Physiological
characterisation of magnesium deficiency in sugar beet: acclimation to
low magnesium differentially affects photosystems I and II. Planta.
220(2):344–355. doi:10.1007/s00425-004-1340-4.
Kashem MDA, Kawai S. 2007. Alleviation of cadmium phytotoxicity by
magnesium in Japanese mustard spinach. Soil Sci Plant Nutr. 53(3):
246–251. doi:10.1111/j.1747-0765.2007.00129.x.
Khan ZI, Safdar H, Ahmad K, Wajid K, Bashir H, Ugulu I, Dogan Y.
2019. Health risk assessment through determining bioaccumulation
of iron in forages grown in soil irrigated with city effluent. Environ
Sci Pollut Res Int. 26(14):14277–14286. doi:10.1007/s11356-019-
04721-1.
Kopsell DA, Kopsell DE. 2006. Accumulation and bioavailability of
dietary carotenoids in vegetable crops. Trends Plant Sci. 11(10):
499–507. doi:10.1016/j.tplants.2006.08.006.
Kosobrukhov A, Knyazeva I, Mudrik V. 2004. Plantago major plants
responses to increase content of lead in soil: growth and
372 S. FAIZ ET AL.
photosynthesis. Plant Growth Regul. 42(2):145–151. doi:10.1023/
B:GROW.0000017490.59607.6b.
Lamhamdi M, Bakrim A, Aarab A, Lafont R, Sayah F. 2011. Lead
phytotoxicity on wheat (Triticum aestivum L.) seed germination and
seedlings growth. C R Biol. 334(2):118–126. doi:10.1016/j.crvi.2010.
12.006.
Masuda T. 2008. Recent overview of the Mg branch of the tetrapyrrole
biosynthesis leading to chlorophylls. Photosynth Res. 96(2):121–143.
doi:10.1007/s11120-008-9291-4.
Mateljan G. 2007. The world’s healthiest food. Nutrient in cabbage,
shredded, boiled. Chicago (IL): George Mateljan Foundation.
Nedim O, Daml BO. 2015. Effect of Mg fertilization on some plant
nutrient interactions and nut quality properties in Turkish hazelnut
(Corylus avellana L.). Sci Res Essays. 10 (14):465–470. doi:10.5897/
SRE2014.5811.
Nejia F, Amine E, Walid Z, Abderrazak S, Chedly A, Mokded R. 2016.
Effects of magnesium deficiency on photosynthesis and carbohydrate
partitioning. Acta Physiol. Plant. 38:145.
Portis ARJ. 2003. Rubisco activase – rubisco’s catalytic chaperone.
Photosyn Res. 75:11–27. doi:10.1023/A:1022458108678.
Puthiyaveetil S, van Oort B, Kirchhoff H. 2017. Surface charge dynam-
ics in photosynthetic membranes and the structural consequences.
Nat Plants. 3:17020.
Rout GR, Das P. 2003. Effect of metal toxicity on plant growth
and metabolism. Agron Sustain Dev. 23(1):3–11. doi:10.1051/
agro:2002073.
Sharma KD, Karki S, Thakur NS, Attri S. 2012. Chemical composition,
functional properties and processing of carrot-a review. J Food Sci
Technol. 49(1):22–32. doi:10.1007/s13197-011-0310-7.
Sharma S, Singh VK, Kumar A, Mallubhotla S. 2019. Effect of nanopar-
ticles on oxidative damage and antioxidant defense system in plants.
Mol Plant Abiotic Stress. 1:315–333.
Singh DP, Beloy J, McInerney JK, Day L. 2012. Impact of boron, cal-
cium and genetic factors on vitamin C, carotenoids, phenolic acids,
anthocyanins and antioxidant capacity of carrots (Daucus carota).
Food Chem. 132(3):1161–1170. doi:10.1016/j.foodchem.2011.11.045.
Strzałka K, Kostecka-Gugała A, Latowski D. 2003. Carotenoids and
environmental stress in plants: significance of carotenoid-mediated
modulation of membrane physical properties. Russ J Plant Physiol.
50(2):168–173. doi:10.1023/A:1022960828050.
Wierzbicka MH, Przedpełska E, Ruzik R, Ouerdane L, Połe
c-Pawlak K,
Jarosz M, Szpunar J, Szakiel A. 2007. Comparison of the toxicity
and distribution of cadmium and lead in plant cells. Protoplasma.
231(1–2):99–111. doi:10.1007/s00709-006-0227-6.
Yasseen BT, Al-Thani RF. 2013. Agricultural chemistry. USA: InTech.
Yoo KS, Bang H, Lee EJ, Crosby K, Patil BS. 2012. Variation of carot-
enoid, sugar, and ascorbic acid concentrations in watermelon geno-
types and genetic analysis. Hortic Environ Biotechnol. 53(6):
552–560. doi:10.1007/s13580-012-0014-6.
Zhang Q, Zhai J, Shao L, Lin W, Peng C. 2019. Accumulation of
anthocyanins: an adaptation strategy of Mikania micrantha to low
temperature in winter. Front Plant Sci. 10:1049. doi:10.3389/fpls.
2019.01049.