Plant Biotechnology Reports (2022) 16:519–528 Online ISSN 1863-5474

https://doi.org/10.1007/s11816-022-00784-8 Print ISSN 1863-5466

ORIGINAL ARTICLE

Comparison of pollution tolerance in sunflowers as a case

study to establish risk assessment criteria for transgenic plants
for environmental remediation
Sung Min Han1 · Kyong‑Hee Nam1

Received: 24 May 2022 / Revised: 26 June 2022 / Accepted: 25 July 2022 / Published online: 10 August 2022
© Korean Society for Plant Biotechnology 2022

Abstract
Transgenic crops for environmental remediation can help in improving polluted areas by increasing plant absorption capac-
ity or resistance to pollutant toxicity. Risk assessment of these crops requires comparative analysis vis-à-vis non-transgenic
counterparts in the possible receiving environments. In this study, we first established a risk assessment environment for
transgenic crops by simulating a heavy metal contamination site in pots using soil from an abandoned mine. Second, we
investigated how traits and heavy metal concentrations changed in plants grown in contaminated and uncontaminated soils
using sunflower as a study system. We found that the concentration of heavy metals in the contaminated soil increased by
3.5–1543.7 times compared with that in the uncontaminated soil. Increased levels of heavy metals in the soil shortened the
flowering period of sunflower plants and caused changes in phenotypic performance despite cultivar differences. Addition-
ally, heavy metal accumulation was significantly increased in the roots and leaves of sunflowers grown on contaminated
soil. Based on these results, we confirmed that the polluted environment was appropriately set to evaluate its effect on the
remediation ability and environmental risk of transgenic crops. Our findings might contribute to developing standards for
risk assessment and safety management of transgenic crops for environmental remediation in the future.

Keywords Bioaccumulation factor · Environmental remediation · Heavy metal · Phytoremediation · Risk assessment ·
Transgenic crop

Abbreviations Introduction
LMO Living modified organisms
MOE Ministry of Environment Environmental pollution is a widespread concern that signif-
ICP-OES Inductively coupled plasma optical emission icantly impacts not only human health but also animals and
spectroscopy plants. As a result, a number of strategies involving chemi-
EC Electrical conductivity cal, physical, and biological methods have been developed to
CEC Cation exchange capacity remediate and recover polluted environments (Macek et al.
FW Fresh weight 2000; Khin et al. 2012; Kemp et al. 2013). Phytoremediation
DW Dry weight is one of them, and it is a rapidly evolving element in envi-
BAF Bioaccumulation factor ronment-friendly and cost-effective technology (Salt et al.
TF Translocation factor 1995; Suresh and Ravishankar 2004) Moreover, the use of
ANOVA Analysis of variance modern biotechnology has provided significant insights into
GLM General linear model the potential of phytoremediation (Pilon-Smits and Pilon
HSD Honestly significant difference 2002; Cherian and Oliberia 2005; Van Aken 2008; Abhilash
et al. 2009).
The development of living modified organisms (LMOs)
* Kyong‑Hee Nam for environmental remediation may contribute to improving
khnam@nie.re.kr polluted environments worldwide because transgenic plants
1 can substantially increase the efficiency of phytoremediation
Division of Ecological Safety, National Institute of Ecology,
1210 Geumgang‑ro, Maeseo‑myeon, Seocheon 33657, to accumulate, transport, and tolerate various types of pollut-
Republic of Korea ants (Flocco et al. 2004; Zhang et al. 2013; Das et al. 2016).

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However, there are concerns that pollutants accumulated in
transgenic plants with enhanced phytoremediation capacity
may have a negative effect by entering into higher levels of
the food chain through those plants (Abhilash et al. 2009;
Kim et al., 2021). Furthermore, pollutant-tolerant transgenic
crops, similar to drought-tolerant transgenic crops, are more
likely to be grown in limited conditions (e.g., contaminated
sites) where wild plants are less likely to survive, allowing
them to invade new habitats and negatively impact biodiver-
sity (Liang 2016). Concerns have also been raised regarding
the disposal of transgenic plants that have accumulated pol-
lutants (Abhilash et al. 2009). Therefore, before commer-
cializing LMOs for environmental remediation, a systematic
evaluation of their risks and a management system for their
safe use should be developed.
The Ministry of Environment (MOE) in South Korea
oversees the development, production, import, export,
distribution, and use of LMOs for environmental remedia-
tion (KBCH 2022; LESC 2022). The National Institute of
Ecology (NIE), under the MOE, has been carrying out risk
assessment studies to develop risk review standards and
safety management system of LMOs for environmental
remediation by operating an institute of LMO risk assess-
ment (Nam and Han 2020; Kim et al. 2021). This risk assess-
ment institute evaluates genetic characteristics of LMOs,
changes in trait characteristics, environmental risks, pro-
duction of hazardous substances, and effects on ecosystem
residues.
The evaluation of LMOs for environmental remediation
requires the establishment of a polluted environment. How-
ever, since most studies on the use of LMOs for environ-
mental remediation are currently conducted at the labora-
tory level, there are only a few reports on the evaluation
of their remediation ability at the actual contaminated site
and their effect on environmental risks (Shim et al., 2013;
Legault et al. 2017). Because demonstrating the remedia-
tion efficiency and risk to the natural environment in the
field is an essential step in the commercial use of LMOs for
environmental remediation, it is important to properly estab-
lish the conditions of a polluted environment and to set up
evaluation criteria for these stress conditions. In this study,
we simulated a heavy metal-contaminated environment in
a pot using soil from an abandoned mine, and examined
the changes in trait characteristics of plants grown in both
contaminated and non-contaminated soils. The findings in
the current study will provide a foundation for large-scale
field experiments and for the development of risk assessment
guidelines for LMOs used in environmental remediation in
the future.
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Plant Biotechnology Reports (2022) 16:519–528
Materials and methods
Plant materials
In this study, sunflowers (Helianthus annuus L.) were used
as test materials prior to using transgenic crops because
they are potential candidates for plant-based remediation
owing to their effective heavy metal hyperaccumulation
(January et al. 2008; Nam and Han 2020). The following
four sunflower cultivars, with different seed properties,
plant height, and germination rates, were selected: Jaer-
aejongja (standard type, height: 1.6–1.8 m), Jaeraejong
(semi-dwarf type, height: 1.0–1.6 m), Jaeraejong1 (dwarf
type, height: 0.5–1.0 m), and Jaeraejong2 (extreme dwarf
type, height: < 0.5 m) (Nam and Han 2020). Seeds were
sown in 40-cell trays filled with commercially available
potting soil (Baroker, Seoul Bio Co., Eumseong, Republic
of Korea) and grown in a confined field (36°01′45.4′′N,
126°43′20.4′′E; elevation: 20 m). Four-weeks-old seed-
lings were transplanted into pots (44 cm diameter × 40 cm
length, 55 L) filled with contaminated or non-contami-
nated soils. In each pot, four sunflower cultivars were
planted sequentially, and all holes in the pot were closed to
prevent the loss of heavy metals. During the experimental
period, the average daily air temperature was 26.9 ± 1.5 °C
and long-day conditions (14 h light/10 h dark) prevailed
(KMA 2022).
Soil sampling and characterization
To establish a heavy metal-contaminated environment, soil
was collected from an abandoned mine area located in
Cheongyang (126°52′47.27″E, 36°31′9.56″N), Republic
of Korea. Commercial potting soil (Baroker, Seoul Bio
Co., Eumseong, Republic of Korea) was used as uncon-
taminated control soil. Air-dried soil was crushed with a
hammer and screened through a 2 mm sieve after removing
stones and plant leaves. The concentrations of As, Cd, Cu,
Ni, Pb, and Zn in powdered soil samples were measured
following digestion with a nitric acid and hydrochloric
acid mixture (3:1, v/v) using inductively coupled plasma
optical emission spectroscopy (ICP-OES; ICPE-9000, Shi-
madzu, Japan) according to the method of the Soil Pollu-
tion Process Test Standards (NIER 2018). The physical
and chemical properties of the soils were analyzed using
the Methods of Soil Chemical Analysis (NIAS 2010).
Briefly, moisture content was determined from the loss in
weight of the samples after oven drying them at 105 °C
until they attained a constant weight. The pH value and
electrical conductivity (EC) of 1:5 soil:water extracts were
measured with an S220 digital pH-meter (Mettler Toledo,
Plant Biotechnology Reports (2022) 16:519–528
Switzerland) and an EC meter (S230, Mettler Toledo),
respectively. The organic matter content was determined
using the Walkley–Black procedure, and total nitrogen and
available phosphorus were determined through the Kjel-
dahl and Lancaster methods, respectively. Cation exchange
capacity (CEC) and exchangeable cations ­(K+, ­Na+, ­Ca2+,
and ­M g 2+) were analyzed using ICP-OES (ICPE-9000,
Shimadzu, Japan) after extraction with ammonium acetate
(1.0 mol/L) at pH 7.0.
Investigation of phenotypic traits in plants
Changes in trait characteristics of plants grown in con-
taminated and non-contaminated soils were measured from
seven days after transplanting. Plant height was measured
from the point where the stem protruded above the soil
to the plant’s tallest point. Stem diameter was measured
with a Vernier caliper at the stem in contact with the soil
surface. Days to flowering were counted from the date of
transplanting to the date when the first signs of flowering
were visible to the naked eye. After four weeks of trans-
planting, plants were gently uprooted from the pots, and
impurities on the roots were removed and washed with
deionized water. The plant samples were then separated
into roots and shoots, and their fresh weight (FW) and
length of both were recorded. The dry weight (DW) of
shoots and roots was determined after oven drying the
samples at 65 °C for five days. The shoot/root ratio was
calculated by dividing the shoot weight of each sample by
its root weight.
Analysis of heavy metal content in plants
For the determination of heavy metal content in plants, dried
samples were divided into roots, stems, and leaves. Each
plant material component was ground, and 0.5 − 1 g portions
of sieved plant matter were digested with nitric acid. The
levels of As, Cd, Cu, and Pb in the digests were determined
using ICP-OES (ICPE-9000, Shimadzu) in accordance with
the procedure stipulated by the Korean Food Code (MFDS
2013).
Bioaccumulation and translocation factors
Bioaccumulation factor (BAF) of As, Cd, Cu, and Pb from
soil to roots was calculated by multiplying the total heavy
metal content of the roots with the total target metal content
of the soil (Alaboudi et al. 2018). Translocation factor (TF),
defined as the ratio of heavy metal concentration in shoots to
that in roots, was measured according to Gupta et al. (2008).
521
Statistical analysis
All results were statistically analyzed using the SAS pack-
age (version 9.1; SAS Institute Inc., Cary, NC, USA) and
STATISTICA (version 8.0; StatSoft Inc., Tulsa, OK, USA).
Student’s t tests were used to evaluate the significance of
differences (p < 0.05) in heavy metal concentrations and
physicochemical properties between non-contaminated and
contaminated soils. Analysis of variance (ANOVA) was used
with a general linear model (GLM) module to compare the
effects of soil, cultivar, and their interactions on phenotypic
characteristics and heavy metal concentrations in plants. Sta-
tistically significant differences between means (p < 0.05)
were identified using the Tukey’s honestly significant differ-
ence (HSD) test. The graphs were plotted using the Sigma-
plot program (version 12.5; Systat Software Inc., San Jose,
CA, USA). Differences in accumulation and translocation
of heavy metals in four sunflower cultivars were examined
via one-way ANOVA, followed by Duncan’s multiple range
tests (p < 0.05).
Results
Soil characteristics and heavy metal concentrations
To compare the phenotypic performance and remediation
ability of plants in a polluted environment, we used heavy-
metal contaminated soil from an abandoned mine and non-
contaminated control soil. The overall levels of heavy metal
differed significantly between non-contaminated and con-
taminated soils, with the As level showing the largest differ-
ence (Table 1). The concentrations of As in the contaminated
soil were variable, ranging from 3,044.1 to 5,841.7 mg ­kg−1,
with mean values 154,269% higher in the contaminated soil
than in the non-contaminated soil. The concentrations of Pb
and Cd were 40,506% and 16,800% higher, respectively, in
the contaminated soil than in the non-contaminated soil. The
Cu, Zn, and Ni levels were increased by 3,524%, 2,124%,
and 252% in the contaminated soil compared with that in the
non-contaminated soil.
Physical and chemical characteristics of soils are listed
in Table 2. Except for total nitrogen and exchangeable C ­ a2+,
the majority of the components differed statistically between
non-contaminated and contaminated soils. Non-contam-
inated soil had significantly higher soil moisture, organic
matter content, and available phosphorus (p < 0.05) than
contaminated soil. The non-contaminated soil was weakly
acidic with a pH of 5.42 ± 0.38, whereas contaminated soil
was weakly alkaline with a pH of 8.52 ± 0.16. Levels of EC,
CEC, and exchangeable K ­ +, ­Na+, and M
­ g2+ were also sig-
nificantly greater (p < 0.05) in non-contaminated soil than
in contaminated soil.
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Table 1  Heavy metal Component Non-contami- Contaminated soil t value Degrees of p value
concentrations (mg ­kg−1) nated soil freedom
in non-contaminated and
contaminated soils As 3.2 ± 2.7 4939.8 ± 986.1 − 12.26 10 < 0.001
Cd 0.1 ± 0.1 16.9 ± 0.8 − 52.35 10 < 0.001
Cu 5.8 ± 0.9 210.2 ± 59.5 − 8.42 10 < 0.001
Ni 11.1 ± 1.2 39.1 ± 3.5 − 18.49 10 < 0.001
Pb 3.3 ± 1.3 1340 ± 346.4 − 9.45 10 < 0.001
Zn 29.3 ± 6.7 651.7 ± 57.2 − 26.46 10 < 0.001

Data are means (n = 6) ± standard deviations

p values are from Student’s t-tests

Table 2  Physical and chemical Component Non-contaminated soil Contaminated soil t value Degrees of p value
properties in non-contaminated freedom
and contaminated soils
Moisture (%) 40.42 ± 12.90 7.59 ± 0.43 5.09 6 0.002
pH 5.42 ± 0.38 8.52 ± 0.16 − 15.03 6 < 0.001
EC (ds ­m−1) 9.29 ± 1.83 0.69 ± 0.29 9.28 6 < 0.001
OC (%) 62.01 ± 0.90 0.01 ± 0.00 138.01 6 < 0.001
TN (%) 0.02 ± 0.01 0.02 ± 0.01 0.78 6 0.467
AP (mg ­kg−1) 253.91 ± 18.21 76.29 ± 14.08 15.43 6 < 0.001
Ca (cmol ­kg−1) 14.51 ± 2.37 11.48 ± 0.97 2.37 6 0.056
K (cmol ­kg−1) 8.63 ± 1.57 0.17 ± 0.02 10.76 6 < 0.001
Mg (cmol k­ g−1) 8.93 ± 1.31 0.55 ± 0.11 12.80 6 < 0.001
Na (cmol k­ g−1) 7.60 ± 2.40 0.09 ± 0.03 6.25 6 0.001
CEC (cmol k­ g−1) 39.67 ± 4.59 12.29 ± 0.96 11.68 6 < 0.001

Data are means (n = 4) ± standard deviations

p values are from Student’s t-tests
EC electrical conductivity, OC organic matter content, TN total nitrogen, AP available phosphorus, CEC
cation exchange capacity

Phenotypic performance of plants grown
under contaminated and non‑contaminated soils
An increase in the content of heavy metals in the soil
delayed the flowering time of sunflower plants (Fig. 1a). In
particular, the Jaeraejongja plants grown in contaminated
soil flowered 7.4 days earlier on average than those grown
in non-contaminated soil; the Jaeraejong plants flowered
11 days earlier than those grown in non-contaminated soil.
In addition, flowers of sunflower plants grown in contami-
nated soil showed a typical stress response owing to mor-
phological changes as shown in Figs. 1b, c.
The effects of heavy metal contamination on pheno-
typic characteristics of different sunflower cultivars were
evaluated by measuring plant height, stem diameter, plant
weight, and plant length (Table 3 and Fig. 2). The final
plant height (12.1%), ratio of the plant height to its initial
value (14.4%), ratio of the stem diameter to its initial value
(11.7%), shoot length (11.6%), shoot FW (23.8%), and the
ratio between shoot and root FW (31.1%) and DW (22.8%)
were significantly lower for plants grown in contaminated
soil than for those grown in non-contaminated soil. The
ratio of plant height to its initial value (< 0.001), final
stem diameter (0.002), shoot length (< 0.001), shoot DW
(< 0.001), root DW (0.002), and ratio of shoot and root FW
(< 0.001) in different cultivars showed statistically sig-
nificant differences. In particular, the final stem diameter
of Jaeraejongja was 39.2% and 35.9% more than that of
Jaeraejong1 and Jaeraejong2, respectively, in the contami-
nated soils. Shoot length of Jaeraejongja and Jaeraejong
was significantly greater than that of Jaeraejong1 and Jaer-
aejong2 both in the non-contaminated and contaminated
soils. Shoot FW of Jaeraejong was 86.9% higher than that
of Jaeraejong1 in the non-contaminated soil. In contrast,
shoot FW of Jaeraejongja and Jaeraejong was more than
that of Jaeraejong1 by 298.3% and 174.8%, respectively.
Shoot DW of Jaeraejongja was significantly higher than
that of the other three cultivars and root DW of Jaerae-
jongja was considerably greater than that of Jaeraejong1 in
the contaminated soils. A significant interaction between

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Fig. 1  Floral characteristics of

different sunflower cultivars
grown in non-contaminated
and contaminated soils. a days
to flowering, b flower of plant
grown in non-contaminated soil
and c flowers of plant grown in
contaminated soils

soil and cultivar was observed for the final shoot diameter
(0.002) and root FW (0.037).

Table 3  Statistical analyses of phenotypic characteristics from differ- Heavy metal accumulation in different plant tissues
ent sunflower cultivars grown in non-contaminated and contaminated
soils
To compare the heavy metal absorption capacity of plants,
Traits p value
the content of heavy metals was estimated in the roots,
Soil treatment Cultivar Soil × Cultivar stems, and leaves of each sunflower cultivar (Table 4 and
Fig. 3). The content of As (2,702.0%), Cd (232.1%), Cu
Plant height (cm)1 0.003 0.212 0.140
(181.5%), and Pb (964.4%) in the roots of plants grown in
Plant height (ratio)2 0.003 < 0.001 0.392
contaminated soil was significantly higher than that in non-
Stem diameter (cm)1 0.591 0.002 0.002
contaminated soil. However, in the stem, only the content
Stem diameter (ratio)2 < 0.001 0.430 0.258
of As was 274.8% higher in plants grown in contaminated
Shoot length (cm) 0.005 < 0.001 0.332
soils than that in non-contaminated soil. In addition, the
Root length (cm) 0.424 0.055 0.905
content of As, Cu, and Pb in the leaves of plants grown in
Shoot FW (g) 0.008 < 0.001 0.082
contaminated soil was significantly increased by 3,902.3%,
Root FW (g) 0.924 0.029 0.037
37.7%, and 162.5%, respectively, when compared with that
Shoot DW (g) 0.771 < 0.001 0.055
in non-contaminated soil.
Root DW (g) 0.178 0.002 0.064
Among the different cultivars, statistical difference was
Shoot/Root (FW) 0.002 0.001 0.269
observed for the content of Cu in stems (0.002), and for
Shoot/Root (DW) 0.048 0.109 0.349
the content of Cd (0.010) and Cu (0.001) in leaves. In par-
Data are means (n = 6) ± standard deviations ticular, the Cu content of stem in Jaeraejong2 was 206.3%
p values calculated using general linear model (GLM)-based method and 149.8% higher than that in Jaeraejongja and Jaeraejong,
FW fresh weight, DW dry weight respectively, in the non-contaminated soil. The Cu content
1
final value
2
ratio of final value to initial value

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Fig. 2  Phenotypic characteristics of different sunflower cultivars median. Different letters above the box indicate significant differences
grown in non-contaminated and contaminated soils. The boundary between means (p < 0.05). Ja, Jaeraejongja; Jg, Jaeraejong; J1, Jaerae-
of the box closest to zero and farthest from zero indicates the 25th jong1; J2, Jaeraejong2; Non-contam, non-contaminated soil; Contam,
and 75th percentiles, respectively, and a line within the box marks the contaminated soil

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Table 4  Statistical analyses of heavy metal concentrations in the “the abandoned mine” is one of the most severely polluted
roots, stems, and leaves from different sunflower cultivars grown in areas. The contaminated soil and water from these areas
non-contaminated and contaminated soils
can cause serious problems for crops and the people who
Component p-value consume them (Jung and Jung 2006). In this study, we used
Soil treatment Cultivar Soil × Cultivar soil from an abandoned mine to recreate the polluted envi-
ronment. We found that the heavy metal concentration in
Root the soil from the mine was 3.5 to 1543.7 times higher than
As < 0.001 0.456 0.427 that in the non-contaminated control soil, depending on the
Cd < 0.001 0.885 0.556 type of heavy metal. Moreover, the As, Cd, and Pb content
Cu < 0.001 0.812 0.640 exceeded the standard for soil contamination, necessitating
Pb < 0.001 0.108 0.277 immediate countermeasures owing to the risk of interfering
Stem with human health and property or rearing of animals and
As 0.009 0.441 0.803 plants (MOE 2018). Therefore, the soil used in this study
Cd 0.120 0.444 0.661 was considered to be similar to the polluted environment
Cu 0.276 0.002 0.271 that will exist when environmental remediation LMOs are
Pb 0.791 0.494 0.077 introduced in South Korea in the future.
Leaf The soil used in this study showed a significant difference
As < 0.001 0.137 0.146 from non-contaminated control soil not only in heavy metal
Cd 0.355 0.010 0.369 content but also in physiochemical properties. Sunflower
Cu 0.001 0.001 0.242 growth and development were also affected by soil differ-
Pb 0.004 0.181 0.082 ences. Exposure to heavy metal stress results in significant
Data are means (n = 6) ± standard deviations biochemical, physiological, and morphological responses
p values calculated using general linear model (GLM)-based method in plants (Gąsecka et al. 2012; Maleci et al. 2014). Excess
heavy metals inhibit plant transpiration and photosynthesis,
interfere with carbohydrate metabolism, and cause nutrient
of Jaeraejong2 leaves was 111.4% higher than that of Jaer- and oxidative stress, all of which affect plant growth and
aejongja in the contaminated soil. Heavy metal content of development (Krämer and Clemens 2005). These changes
roots, stems, and leaves showed no significant difference for eventually result in visible symptoms such as growth inhibi-
the different soil and cultivar types. tion, chlorosis, root browning, withering, and death (Ozturk
et al. 2008; Ghori et al. 2019). In this study, rapid flower-
Accumulation and translocation of heavy metals ing and morphological changes were observed in sunflower
in sunflower cultivars plants grown in contaminated soils, confirming that the
plants experienced stress. In addition, the phenotypic char-
The BAF and TF values for As, Cd, Cu, and Pb are shown in acteristics such as plant height, stem diameter, shoot length,
Fig. 4. The transfer of Cd and Cu from soil into plant roots FW, ratio of shoot/root FW, and DW were reduced by heavy
was higher than that of As and Pb in all sunflower cultivars. metals, although there were cultivar differences. Change
However, all values were recorded to be less than 1. Also, in trait characteristics of LMOs is one of the main factors
no significant difference was observed between cultivars. evaluated in the institute of LMO risk assessment for envi-
Differences in the transfer of heavy metals from plant roots ronmental remediation in South Korea. Such an assessment
to shoots between sunflower cultivars were only observed of the expression characteristics of introduced LMOs in an
for Pb. The TF of Pb was significantly higher in Jaeraejong acceptable contaminated environment could also provide
1 than in other sunflower cultivars. relevant information for environmental risks such as their
weediness and invasiveness potential.
Biotechnological approaches for environmental remedia-
Discussion tion are associated with increasing specific plant absorp-
tion capacity or resistance to heavy metal toxicity (Ovečka
For environmental remediation, LMOs can be used to and Takáč 2014; Ghori et al. 2019). Expression of certain
improve contaminated sites worldwide. However, polluted traits (such as stress tolerance) in transgenic plants under
environments vary according to industrial status, policies, actual stress conditions can cause changes in the chemical
cultures, and the location of countries. In South Korea, composition of the plants, affecting the surrounding eco-
there are various contaminated sites including industrial system, including microorganisms and insects (Conner
complexes, waste landfills, mines, smelting areas, and et al. 2003; Nam et al. 2014, 2016). In particular, higher
transportation-related facilities (SGIS 2022). Among them, absorption of heavy metals in transgenic plants could lead

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Fig. 3  Heavy metal concentrations in different parts of sunflower cul-
tivars grown in non-contaminated and contaminated soils. The con-
centrations are given in mg ­kg−1. The boundary of the box closest to
zero and farthest from zero indicates the 25th and 75th percentile,
respectively, and a line within the box marks the median. Different
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Plant Biotechnology Reports (2022) 16:519–528
letters above the box indicate significant differences between means
(p < 0.05). Ja, Jaeraejongja; Jg, Jaeraejong; J1, Jaeraejong1; J2, Jaer-
aejong2; Non-contam, non-contaminated soil; Contam, contaminated
soil
Plant Biotechnology Reports (2022) 16:519–528 527

Fig. 4  Accumulation and trans-

location of As, Cd, Cu, and Pb
in different sunflower cultivars.
Data are presented as means
(n = 5) ± standard deviations.
p values are from one-way
analysis of variance (ANOVA).
Different letters indicate signifi-
cant differences between means
using Duncan’s multiple range
test (p < 0.05)

to bioaccumulation at higher trophic levels in a food chain,
resulting in ecosystem changes (Kim et al. 2021). In this
study, heavy metal accumulation greatly increased in the
roots and leaves of sunflower grown in contaminated soils,
but not in the stems, except for As. In addition, most of the
BAF and TF values did not differ significantly among sun-
flower cultivars. Based on these results, it was confirmed
that a contaminated environment for the evaluation of the
remediation ability of transgenic plants and their impact on
environmental risks was successfully established.
Adoption of LMOs will be contingent on appropriate
safety assessments and public acceptance (Liang 2016).
Before introducing LMOs for environmental remediation,
it is especially important to carefully consider the risks
to the natural ecosystem because they can grow under the
restricted conditions of contaminated sites or cause translo-
cation of pollutants to the surrounding ecosystem. Therefore,
comparative analysis with non-transgenic counterparts in
the possible receiving environment to be grown, is required.
Although there is currently a harmonized approach to risk
assessment of LMOs around the world, there is little infor-
mation about risk assessment of LMOs for environmental
remediation. Here, we compared pollution tolerance in
various sunflower cultivars as a case study using contami-
nated mine soils, prior to the risk assessment of LMOs for
environmental remediation. Our results will provide impor-
tant insights for field trials to prepare risk assessment items
and standards of LMOs for environmental remediation in
South Korea.
Funding This research was supported by the National Institute of Ecol-
ogy (NIE) and funded by the Ministry of Environment (MOE) of the
Republic of Korea (NIE-A-2022–04).
Declarations
Conflict of interest The authors declare that they have no competing
interests.
References
Abhilash PC, Jamil S, Singh N (2009) Transgenic plants for enhanced
biodegradation and phytoremediation of organic xenobiotics. Bio-
technol Adv 27:474–488
Alaboudi KA, Ahmed B, Brodie G (2018) Phytoremediation o Pb and
Cd contaminated soils by using sunflower (Helianthus annuus)
plant. Ann Agric Sci 63:123–127

13
528
Cherian S, Oliberia MM (2005) Transgenic plants in phytoremedia-
tion: recent advances and new possibilities. Environ Sci Technol
39:9377–9390
Conner AJ, Glare TR, Nap JP (2003) The release of genetically modi-
fied crops into the environment. Plant J 33:19–46
Das N, Bhattacharya B, Maiti MK (2016) Enhanced cadmium accu-
mulation and tolerance in transgenic tobacco overexpressing rice
metal tolerance protein gene OsMTP1 is promising for phytore-
mediation. Plant Physiol Biochem 105:297–309
Flocco CG, Lindblom SD, Pilon Smits EAH (2004) Overexpression of
enzymes involved in glutathione synthesis enhances tolerance to organic
pollutants in Brassica juncea. Int J Phytoremediation 6:289–304
Gąsecka M, Mleczek M, Drzewiceka K, Magdziak Z, Rissmann I,
Chadzinikolau T, Golinski P (2012) Physiological and morpho-
logical changes in Salix viminalis L. as a result of plant exposure
to copper. J Environ Sci Health A Tox Hazard Subst Environ Eng
47:548–557
Ghori NH, Ghori T, Hayat MQ, Imadi SR, Gul A, Ozturk M (2019)
Heavy metal stress and responses in plants. Int J Environ Sci Tech-
nol 16:1807–1828
Gupta S, Nayek S, Saha RN, Satpati, (2008) Assessment of heavy metal
accumulation in macrophyte, agricultural soil, and crop plants
adjacent to discharge zone of sponge iron factory. Environ Geol
55:731–739
January MC, Cutright TJ, Van Keulen H, Wei R (2008) Hydroponic
phytoremediation of Cd, Cr, Ni, As, and Fe: Can Helianthus
annuus hyperaccumulate multiple heavy metals? Chemosphere
70:531–537
Jung MC, Jung MY (2006) Evaluation and management method of
environmental contamination from abandoned metal mines in
Korea. J Korean Soc Geosystem Eng 43:383–394
KBCH (2022) Law and System; Korea Biosafety Clearing House: Dae-
jeon, Korea. Available online: https://​www.​biosa​fety.​or.​kr/​portal/​
index.​do?​pageid=​c_​01.​Acces​sed 4 Mar 2022
Kim DE, Kim D, Ban YG, Lee M, Lee H, Jo A, Han SM, Lee JR,
Nam KH (2021) Analysis of arthropod communities in sunflower-
cultivated fields to develop risk assessment guidelines for LMO
used for environmental remediation. PNIE 2:129–138
Kemp KC, Seema H, Saleh M, Le NH, Mahesh K, Chandra V, Kim KS
(2013) Environmental applications using graphene composites:
water remediation and gas adsorption. Nanoscale 5:3149–3171
Khin MM, Nair AS, Baub VJ, Murugan R, Ramakrishna S (2012) A
review on nanomaterials for environmental remediation. Energy
Environ Sci 5:8075–8109
KMA (2022) Synoptic weather observation, statistics by condition;
Korea Meteorological Administration: Seoul, Korea. Available
online: https://​data.​kma.​go.​kr Accessed 4 Mar 2022
Krämer U, Clemens S (2005) Functions and homeostasis of zinc, cop-
per, and nickel in plants. In: Tamas M, Martinoia E (eds) Topics
in Current Genetics. Springer, Heidelberg, pp 215–271
Legault EK, James CA, Stewart K, Muiznieks I, Doty SL, Strand SE
(2017) A field trial of TCE phytoremediation by genetically modi-
fied poplars expressing cytochrome P450 2E1. Environ Sci Tech-
nol 51:6090–6099
LESC (2022) LMO Safety Management System; LMO Environmental
Safety Center: Seocheon, Korea. Available online: http://​lesc.​nie.​
re.​kr/​lesc/​conte​nts.​do?​cid=​3&​idx=9 Accessed 4 Mar 2022
Liang C (2016) Genetically modified crops with drought tolerance:
achievements, challenges, and perspectives. Drought Stress Toler-
ance in Plants vol 2, Springer, Cham, pp 531–547
Macek T, Macková M, Káš J (2000) Exploitation of plants for the
removal of organics in environmental remediation. Biotechnol
Adv 18:23–34
Maleci L, Buffa G, Wahsha M, Bini C (2014) Morphological changes
induced by heavy metals in dandelion (Taraxacum officinale
Web.) growing on mine soils. J Soils Sediments 14:731–743
13
Plant Biotechnology Reports (2022) 16:519–528
MFDS (2013) Food code. Ministry of Food and Drug Safety, Cheongju,
Korea
MOE. Soil Environment Conservation Act (Act No. 15658 of Jun. 12,
2018); Ministry of Environment: Sejong-si, Korea, 2017. Avail-
able online: https://e​ law.k​ lri.r​ e.k​ r/k​ or_s​ ervic​ e/l​ awVie​ w.d​ o?h​ seq=​
48720​&​lang=​ENG Accessed 4 Mar 2022
Nam KH, Han SM (2020) Seed germination of sunflower as a case
study for the risk assessment and management of transgenic plants
used for environmental remediation in South Korea. Sustainability
12(23):10110
Nam KH, Kim DY, Shin HJ, Nam KJ, An JH, Pact IS, Park JH, Jeong
SC, Kim HB, Kim CG (2014) Drought stress-induced composi-
tional changes in tolerant transgenic rice and its wild type. Food
Chem 153:145–150
Nam KH, Shin HJ, Pack IS, Park JH, Kim HB, Kim CG (2016) Metab-
olomic changes in grains of well-watered and drought-stressed
transgenic rice. J Sci Food Agric 96:807–814
NIAS (2010) Methods of Soil Chemical Analysis. Sam-Mi press,
National Institute of Agricultural Sciences, pp 13–290
NIER (2018) Soil pollution process test standards. National Institute
of Environmental Research, Incheon, Korea
Ovečka M, Takáč T (2014) Managing heavy metal toxicity stress in
plants: biological and biotechnological tools. Biotechnol Adv
32:73–86
Ozturk M, Yucel E, Gucel S, Sakcali S, Aksoy A (2008) Plant as bio-
monitors of trace elements pollution in soil. In: Prasad MNV (ed)
Trace elements: environmental contamination, nutritional benefits
and health implications. Wiley, New York, pp 723–744
Pilon-Smits E, Pilon M (2002) Phytoremediation of metals using trans-
genic plants. Crit Rev Plant Sci 21:439–456
Salt DE, Blaylock M, Kumar NPBA, Dushenkov V, Ensley BD, Chet
I, Raskin I (1995) Phytoremediation: a novel strategy for the
removal of toxic metals from the environment using plants. Bio-
technol 13:468–474
SGIS (2022) Soil Pollution Survey; Soil Groundwater Information Sys-
tem: Sejong, Korea. Available online: https://​sgis.​nier.​go.​kr/​web/​
soilP​ollut​ion?​pMENU_​NO=​17 Accessed 4 Mar 2022)
Shim D, Kim S, Choi YI, Song WY, Park J, Youk ES, Jeong SC, Mar-
tinoia E, Noh EW, Lee Y (2013) Transgenic poplar trees express-
ing yeast cadmium factor 1 exhibit the characteristics necessary
for the phytoremediation of mine tailing soil. Chemosphere
90:1478–1486
Suresh B, Ravishankar GA (2004) Phytoremediation−a novel and
promising approach for environmental clean-up. Crit Rev Plant
Sci 24:97–124
Van Aken B (2008) Transgenic plants for phytoremediation: helping
nature to clean up environmental pollution. Trends Biotechnol
26:225–227
Zhang Y, Liu J, Zhou Y, Gong T, Wang J, Ge Y (2013) Enhanced
phytoremediation of mixed heavy metal (mercury)−organic pol-
lutants (trichloroethylene) with transgenic alfalfa co-expressing
glutathione S-transferase and human P450 2El. J Hazard Mater
260:1100–1107
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