eJournal of Applied Forest Ecology (eJAFE), Vol 9, No.

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ISSN: 2347-4009
Vol-09 Issue-01

International Journal of Applied

Forest Ecological Research
This Journal is online
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Assessment of Cu accumulation in Lepidium sativum L.

Snejana Dineva1*, Mariyana Lyubenova2, IrinaKaradjova3
Faculty of Techniques and Technology, Trakia University - Stara Zagora;
1

Yambol 8602, "Gr. Ignatiev" str. №38, Bulgaria

Faculty of Biology, University of Sofia, 8 D. TzankovBlv. Bulgaria
2

Faculty of Chemistry, Sofia University “St.st. KlimentOhridski”

3

Received on March 22,2021;Accepted on April,12,2021.

Abstract
The aim of the current study is to evaluate Cu++accumulation levels in Lepidium sativum L. growing in different media, as
distilled water (DW)with +125 and +250 mg.l-1 added copper ions, and purified industrial wastewater (PIWW), with the same
concentrations of copper ions in addition. It was registered that the accumulation of Cu2+ in Lepidium sativum L. depended from
the concentration of ions in the water media, as double increase of Cu2+ reflected to approximately three times more accumulation
in the tissues of control samples, while in IPWW the effect was only double bioaccumulation. The excess of Cu2+ions in media
did not influenced on the accumulation of Mn ions. In media from purified wastewater, the addition of copper ions diminished the
uptake of Fe ions, and increased Zn bioaccumulation.
Keywords:Lepidium sativum L., Cu++ accumulation, phytoremediation.

Introduction andhigh efficiency, it is anenvironmentally and eco-

friendly resolution based on solar energy and the use of
Heavy metals are widespread pollutants, causing
plant species(Mojiri et al., 2012; Ali et al., 2013;
harm on the ecosystems and human health (Briffa et al.,
S u m i a h a d i a n d A c a r, 2 0 1 8 ; C i o i c a , e t a l . ,
2020), tending to increase their concentration in an
2019).According to Freeman et al. (2004), nowadays,
organism over the time (Verma and Dwivedi, 2013).
more than 400 plant species are well known as good
High levels of heavy metals are released from
hyper accumulators of various metals (Cd, Co, Cu, Mn,
manufacturing sectors (Ayturan et al., 2018), mining
Ni, and Zn), metalloids (As) and non-metals (Se).Thus,
activities, and smelters, dispersed into nearby
many plants have been introduced for phytoremediation
agricultural soils and streams, taken up by food crops
of heavy metal pollution (Mojiri, 2011;Cruz-Landero et
(P´erez-Sirvent et al., 2007).
al., 20100;Jadia and Fulekar, 2009; Chehregani et al.,
The common heavy metals mentioned as pollutants in
2009;Zhang et al.,2009;Xiao et al., 2008;Subroto et al.,
the environment are As, Cd, Cr, Cu, Hg, Pb and Ni
2007;Al-Farraj and Al-Wabel, 2007;Ok and Kim,
(Raskin et al., 1997; Lasat, 2000; Sharma, 2013;
2007;Cho-Ruk et al., 2006;Lombi et al., 2001).
Masindi and Muedi, 2018; Briffa et al., 2020). Some of
The aim of research is to evaluate the levels of Cu ++
these metals are micronutrients essential for plant
accumulation in Lepidium sativum L. growing in
growth and development, such as Zn, Cu, Mn, Ni, and
different media, distilled water (DW) and purified
Co, while others did not have any biological function,
industrial wastewater (PIWW), with addition of two
such as Cd, Pb, and Hg (Gaur and Adholeya, 2004).
different concentrations of copper ions 125 ppm and
Especially, three heavy metals Cu, Pb and Zn are
250 ppm.
accepted as very problematic pollutants(Ayturan et al.,
2018). Lepidium sativum L.
Restoration of water and soils polluted with
Lepidium sativum L. is a fast-growing annual herb
potentially toxic metals and metalloids is of a major
belonging to the Brassicaceae family, native to Egypt
global concern (Shelmerdine et al., 2009), because that
and west Asia, widely cultivated in temperate climates
kind of contamination is stable and not destroyed by
all over the world (Neidi et al., 2012). Garden cress can
degradation (Sumiahadi and Acar, 2018).The promising
grow in any type of climate and soil condition (Wadhwa
and noveltechnology for restoration of polluted water
et al., 2012; Nehdi et al., 2012). According to
and lands byremoval the heavy metalsis a
Smolinska and Szczodrowska (2017), with the short
phytoremediation or bioremediation (Mojiri, et al.,
vegetation period and the ability to accumulate heavy
2012; Dixit, et al., 2015). Phytoremediation is a
metals makes Lepidium sativum L. suitable for use in
preferable green technology, because of its low cost

* Corresponding author : Snejana Dineva

email-sbdineva@abv.bg
17
repeated phytoextraction, with harvesting several times
throughout the year. Lepidium sativum L. showed the
capacity to be used in biological soil reclamation
processes (Kathi and Khan, 2011; Gunduz et al., 2012;
Kiayee et al., 2012; Mojiri et al., 2012; Pures and Ihlam,
2014; Vakili and Aboutorab, 2013; Smolinska and
Szczodrowska, 2017; Smolinska, 2019). Lepidium
sativum L. growing well on polluted soil and showed
ability for Ni (Mojiri et al. 2012), Cd, Se and Hg
accumulation (Gill and Tuteja, 2010; Elguera et al.,
2013; Smolinska, 2015; Smolinska and Szczodrowska,
2017).
Garden cress is mentioned as a hyperaccumulating
plant suitable for phytoremediation of soils
contaminated with As, Cd, Pb (Gunduz et al., 2012). In
the study for examination of metals accumulation in
Lepidium sativum L. from irrigated with wastewater
soil, the enrichment factor of the heavy metals in the
edible parts of L. sativum L. followed the order: Cu > Pb
> Cd > Ni > Cr, with a high transfer factor (TF > 1) in the
edible parts of plant for Cd (Keser, 2013).
Materials and Methods
Purified industrial wastewater-The purified
wastewater was collected from Manufacturing Units
that produce wide range of rolled and extruded products
made from copper, copper alloy and zinc. The concern
operates in accordance with rigorous management
systems for environmental protection and occupational
health - ISO 9001; ISO 14001; and OHSAS 18001.
Biotest with Lepidium sativum L. (Garden Cress)-
The accumulation of Cu2+ by Lepidium sativum L. from
different media was accessed. The tests were carried out
on a solid substrate in pots, on floating hydrocracker
stands and in Petri dishes with soaked filter paper
eJournal of Applied Forest Ecology (eJAFE)
Snejana Dineva, Mariyana Lyubenova, IrinaKaradjova
(Lyubenova et al., 2019).
The preliminary preparation of the tests included
preparing the variants: control distilled water (DW);
DW with +125 and + 250 Cu+2ppm; purified industrial
wastewater (PIWW) with +125 and + 250 Cu +2
ppm.The filter paper on the lid of the plate was soaked in
5ml water from different samples. The seeds were kept
in a refrigerator for 24 hours, and then they were soaked
in the distilled water for 24 hours, before starting the
experiment (Lyubenova and Kalchev, 2000).
Experimental Analysis
Flame AAS measurements The determination of
chemical elements was carried out using a flame atomic
absorption spectrometry using Perkin Elmer 400 in
air/acetyleneflame, under optimal instrumental
parameters ensuring maximumsignal-to-noise ratio.
Plants sample digestion
High-pressure laboratory microwave oven (Milestone
Ethos 1600 MicrowaveLabstation, Sorisole, Italy)
operating at a frequency of 2450 Hz, withan energy
output of 900 W was used for microwave digestion of
plantsamples.
Accurately weighted samples around 0.3-0.4 g were
transferredin 50 ml teflon vessels of MW system,
mixture of 3 ml concentratednitric acid and 1 ml
hydrogen peroxide were added, and samples were
leftfor 24 h at room temperature. Wet digestion was
performed in three successive steps:
1. Temperature 90˚ C at maximum power 750 W for 10
min;
2. Cooling and second step at temperature180 at
maximum power 1000 W for 15min;
3. After the digestion procedure is ended and
Assessment of Cu accumulation in Lepidium sativum L.
subsequent cooling, thedigested samples were
transferred in volumetric flask and diluted to afinal
volume of 25.0 mL with water.
Two blank samples were prepared with each run.
Statistical approach
One-way ANOVA was performed using Statistic 7.0
software (Stat soft; www.statsoft.com) to compare the
means of different treatments in each of the tests. The
significance level was set at 0.05.
Results and Discussion
Copper (Cu), zinc (Zn) iron (Fe) and manganese (Mn)
are essential micronutrients to the growth of plants. The
accumulation of those ions from purified industrial
water are presented on figure1 and table 1, there are can
see the enhancement of Fe ions bioaccumulation in
plant tissues and diminishing Cu uptake from the
PIWW medium.
According to Bloodnick (2021a), the ideal range of
Table 1. Accumulation of metal ions in Lepidium sativum L.
Bioaccumulation of Cu2+ in Lepidium sativum Lis
strongly dependantfrom the concentration of ions in the
media, as the positive correlation was found for both
media. It was registered that with the double increase of
Cu2+ in the medium prepared from distillate water (DW)
+125 ppm to DW+250 ppm, the copper accumulation in
the tissues rise up approximately 3 times, from 1157±34
to 3846±113 ppm respectively. If compare the
bioaccumulation with the control samples, then the
18
iron in the tissue should be 20 times higher than that of
copper for standing plant development. In the plant
tissues of Lepidium sativum L. the mean amount of Fe
was 230±9 ppm that is around 7 times higher than that of
Cu ions, 33±2 ppm. According to Owen and Kissel
(2021), the normal range of Cu in many plants varied
from 5 to 20 ppm, with critical values between 3 to 10
ppm. When the Cu concentration in plants is below than
3 ppm, deficiencies are probable to occur, when Cu
levels exceed 20 ppm, toxicities may arise. In most plant
tissues, the range for copper varies between 3-10 ppm
(Bloodnick, 2021a).
The normal range of Zn in most plants is between 20
to 100 ppm, and zinc deficiencies occur when the leaf
level drops below 15 ppm (Owen and Kissel, 2021).
Zinc toxicity can occur when its levels in the tissue
exceed 200 ppm (Bloodnick, 2021b). From table 1, and
figure1, can see that the uptake of Zn ions are not
affected from the different media in both control
samples, staying stable between 175±8 and 182±14
ppm.
increasing of copper in tissues were 31 times more at
concentration +125 ppm, and more than 103 times,
when was added 250 ppm Cu in the media of DW (table
1). When the media was prepared from the PIWW, the
registered effects were a double bioaccumulation of Cu
between different copper concentrations and likening
with the control samples 26 times more at +125 ppm,
and 53 times higher at +250 ppm Cu2+(table 1; figure
2b).
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19
Figure 2. Bioaccumulation of Cu2+ from two different media.
(a) Distilled water (DW) and purified industrial waste
water (PIWW) with 125 ppm Cu++;(b)Different media
and Cu-ion concentrations
The similar trend of strongly positive correlation of
bioaccumulation in plant tissues of Lepidium sativum
Land concentrations of metal ions in media werefound
from Robinson et al.(2003), for As (r=0.96, P<0.001)
and for Pb from soil, as the accumulation increases with
increasing lead concentration to 300 ppm (Pures and
Ihlam, 2014).
According to Bloodnick (2021a), too much copper in
media can lead up to a deficiency of another
micronutrient, as a copper can compete with plant
uptake of iron and sometimes molybdenum or zinc. At
+250 ppm in purified wastewater the low uptake of all
controlled ions occur (table 1), for Zn the registered
bioaccumulation was with mean 117±9 vs 182±14 ppm
and for Mn 29±2 ppm vs 36±2 ppm. The excess of
Cu2+ions in media did not have strongly influence only
on the uptake of Mn ions (table 1, figure 2).
When the media was from PIWW, additional copper
ions diminish the uptake of Fe ions, from 230±9 ppm to
94±4 ppm regardless of the excess concentration (table
1, figure 2). Moreover, one exception was registered,
when to the PIWW have been added +125 ppm Cu2+
then the increasing of Zn concentration in plant tissues
rise up to 226±10 vs 182±14 ppm. Synergetic effect of
Cu and Zn uptake from treated wastewater in Triticum
aestivum L. was reported from Tani and Barrington
(2005).
The amount of Cu ions accumulated in plant tissues
growing in different media strongly correlates with the
increase in concentration (figure2). The PIWW
mitigate growth inhibitory effect of high copper ions
concentrations on Lepidium sativum L. seeds that was
observed earlier from our previous study Lyubenova et
al. (2019), perhaps by lessening its amount involving
free Cu+2 in complexes with other waste products. The
PIWW led to reducing toxicity, and respectively,
eJournal of Applied Forest Ecology (eJAFE)
Snejana Dineva, Mariyana Lyubenova, IrinaKaradjova
decreasing the inhibitory effect of high copper ions
concentration resulted in stimulation of roots and stem's
growth (Lyubenova et al., 2019).
The results from conducted experiment confirm the
ability of watercress to accumulate high levels of Cu++
ions from the medium and enable the plant to be utilized
for phytoremediation.
Conclusions
The study revealed high capacity of watercress to
bioaccumulate copper. It was registered that the
accumulation of Cu2+ in Lepidium sativum L. depended
from the concentrations of ions in media, as double
increase of Cu2+ reflected to approximately three times
more accumulation in the tissues, while in treated water
the effect was only double bioaccumulation. That
indicates the potential of plant to be used for
phytoremediation of contaminated soils or water.
In addition, from the results it can be concluded that
the treated water from the manufacture can be utilize for
irrigation of agriculture land.
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