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Evaluation of growth and biochemical indicators of Salvinia natans exposed to

zinc oxide nanoparticles and zinc accumulation in plants

Article in Environmental Science and Pollution Research · July 2013

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Environ Sci Pollut Res (2014) 21:732–739
DOI 10.1007/s11356-013-1970-9
RESEARCH ARTICLE
Evaluation of growth and biochemical indicators of Salvinia
natans exposed to zinc oxide nanoparticles and zinc
accumulation in plants
Changwei Hu & Xu Liu & Xiuling Li & Yongjun Zhao
Received: 13 March 2013 / Accepted: 28 June 2013 / Published online: 17 July 2013
# Springer-Verlag Berlin Heidelberg 2013
Abstract The adverse effects of zinc oxide nanoparticles
(ZnO NPs) with an average diameter of 25 nm on the aquatic
plant Salvinia natans (L.) All. were determined. Growth,
superoxide dismutase (SOD) activity, catalase (CAT) activity,
peroxidase activity, and chlorophyll content of the plants were
measured after 7 days of exposure to different concentrations
of ZnO NPs (1 to 50 mg L−1). The particle distribution in the
culture medium (without plants) during the first 24 h was
determined using a Nanotrac 250 particle analyzer. We also
investigated the zinc accumulation in leaves and roots of the
plant after 7 days of exposure. Exposure to 50 mg L−1 ZnO
NPs significantly increased SOD and CAT activities (P<0.05)
and significantly depressed photosynthetic pigments
(P<0.05). However, plant growth was not significantly affect-
ed (P>0.05). NPs completely precipitated at the bottom of the
container at 8 h except for the portions of dissolution and
aggregation on the roots. ZnO NPs at a concentration of
50 mg L−1 can adversely affect S. natans, and their stress is
affected by their aggregation and dissolution.
Keywords ZnO nanoparticles . Salvinia natans .
Antioxidant enzymes . Aggregation . Accumulation
Responsible editor: Elena Maestri
C. Hu (*) : X. Li
Shandong Provincial Key Laboratory of Water and Soil
Conservation & Environmental Protection, Linyi University,
Linyi 276000, China
e-mail: changwei.hu@163.com
X. Liu
School of Life Science, Linyi University, Linyi 276000, China
Y. Zhao
College of Biological, Chemical Science and Engineering,
Jiaxing University, Jiaxing 314001, China
Introduction
Nanoparticles (NPs) are materials with at least two dimen-
sions under 100 nm (Masciangioli and Zhang 2003). Their
unique physicochemical properties, such as small size,
chemical composition, surface structure, solubility, shape,
and aggregation, are attractive for a wide range of novel
applications in electronics, healthcare, cosmetics, technolo-
gies, and engineering industries (Lecoanet et al. 2004;
Lecoanet and Wiesner 2004; Sun et al. 2005; Nel et al.
2006). Based on a recent estimate, the number of consumer
products on the market containing NPs or nanofibers ex-
ceeds 1,300 and is still rapidly increasing (Love et al. 2012).
According to “The Nanotechnology Consumer Products
Inventory” (Maynard and Michelson 2006), the most com-
mon metal oxide NP material mentioned in the product de-
scriptions is titanium dioxide (TiO2), followed by zinc oxide
(ZnO), which are widely used in pigments, semiconductors,
sunscreens, and food additives (Handy et al. 2008). These
NPs may be released to the environment, threatening the
ecosystem during the life cycle of these commercial prod-
ucts, although little is known about their actual concentration
in the environment. Gottschalk et al. (2009) reported that
modeled concentrations of ZnO NPs were approximately
0.01 μg L−1 in surface water and 0.432 μg L−1 in sewage
treatment plant effluents in Europe. Boxall et al. (2007) gave
an estimate concentration (less than 100 μg L−1) of ZnO NPs
in water in the UK.
The extensive use of ZnO NPs will undoubtedly increase
the potential risk for human and environmental exposures.
To date, investigations on the ecotoxicology of ZnO NPs are
emerging, which aim to assess their harmful effects to the
ecosystem. Many studies focused on the toxicity of the NPs
on aquatic organisms because the water environment is
regarded as the ultimate destination of NPs released in the
environment (Nel et al. 2006; Kahru and Dubourguier 2010;
Peralta-Videa et al. 2011).
Environ Sci Pollut Res (2014) 21:732–739
The ecotoxicological effects of ZnO NPs on various aquat-
ic organisms are reported (Ma et al. 2013). Meanwhile, the
actual mechanisms of the toxicity need to be better under-
stood. Microalgae are an important model organism for aquat-
ic toxicity assay. Microalga Pseudokirchneriella subcapitata
was sensitive to ZnO NP stress, and 72 IC50 was found to be
less than 70 μg Zn L−1 (Aruoja et al. 2009; Franklin et al.
2007). Miller et al. (2010) reported that ZnO NPs (20 to
30 nm) at 1 mg L −1 inhibited growth of four marine
microalgae including Skeletonema marinoi, Thalassiosira
pseudonana, Dunaliella tertiolecta, and Isochrysis galbana.
All the three aforementioned studies demonstrated that the
toxicity was attributed to solubilized zinc ions. Brayner et al.
(2010) investigated the adverse effects of ZnO NPs to
cyanobacteria Anabaena flos-aquae and found that the pho-
tosynthetic activity of microalgae was inhibited.
Aquatic invertebrates and vertebrates, such as Daphnia
magna and zebrafish Danio rerio, were also selected as test
organisms to evaluate the toxicity of ZnO NPs. Heinlaan
et al. (2008) reported acute toxicity of ZnO NPs on D.
magna. The results showed that the value of 48-h LC50
was 3.2 mg L−1 and the toxicity was due to solubilized Zn
ions. A recent report also indicated that dissolved Zn2+
released from ZnO NP suspension contributed to the toxicity,
and then, aggregation and sedimentation of NPs reduced
their toxicity (Yu et al. 2011). Wiench et al. (2009) found
that the toxicity of ZnO on D. magna was independent of
particle size, coating of particles, aggregation of particles,
and type of medium or the applied pretreatment of the test
dispersions. Zhu et al. (2008) reported that 96 h LC50 of
ZnO NPs to zebrafish was 1.79 mg L−1 and the dissolved
zinc ion was partially responsible for the toxicity. Micro-
sized ZnO NP aggregates in culture medium caused dose-
dependent effects to zebrafish embryo hatching (Zhu et al.
2009). The toxicity was likely due to combined effects of
dissolved zinc ions and particle aggregates.
Although a number of studies on the toxicity of ZnO NPs
were reported, knowledge on their toxicity to aquatic plants
remains lacking. For other NPs, such as silver, copper oxide,
and TiO2, duckweeds (Lemna minor, Spirodela polyrhiza, and
Landoltia punctata) were used for the evaluation of their
toxicity (Gubbins et al. 2011; Kim et al. 2011; Shi et al.
2011; Jiang et al. 2012). However, no reports on other aquatic
plants could be found as yet. Salvinia natans is a free-floating,
aquatic heterosporous fern that reproduces vegetatively and
rapidly forms expanding mats of foliage on still water surfaces
in tropical and subtropical regions. This plant was previously
selected as a test organism to study the toxicity of heavy
metals (Dhir et al. 2009). In addition, environmental behaviors
of NPs such as aggregation and dissolution are regarded as
important aspects in understanding their toxicity (Sharma
2009; Petosa et al. 2010) and need further investigation in
toxicity assay of ZnO NPs to aquatic plants.
733
The present study aimed to evaluate the responses of
growth, photosynthetic pigments, and antioxidant enzyme
activities of S. natans to ZnO NPs. The influence of aggre-
gation on the adverse effects of these NPs and bioaccumu-
lation of zinc in the plants were also investigated.
Materials and methods
Characterization of ZnO NPs
ZnO NPs without coating were purchased from Aipurui
Co., Ltd. (Nanjing, China). The surface area of NPs was
determined using the multipoint Brunauer–Emmett–Teller
(BET) method (Brunauer et al. 1938). The morphology
of the NPs was examined under a transmission electron
microscope (H-7500, Hitachi, Japan).
Incubation and growth of S. natans
Samples of S. natans (L.) All. were collected from the Beng
River (Linyi, China). According to the guidelines of the
Organization for Economic Cooperation and Development
(OECD 2006), the nutrient solution used for growth was
prepared using the following contents (in milligrams per liter):
NaNO3, 85; KH2PO4, 13.4; MgSO4·7H2O, 75; CaCl2·2H2O,
36; Na 2 CO 3 , 20; H 3 BO 3 , 1.0; MnCl 2 ·4H 2 O, 0.20;
Na2MoO4·2H2O, 0.010; ZnSO4·7H2O, 0.005; CuSO4·5H2O,
0.0050; Co(NO3)2·6H2O, 0.010; FeCl3·6H2O, 0.84; and
Na2-EDTA·2H2O, 1.4. pH was adjusted to 6.5±0.2 by adding
0.1 N NaOH.
The plants were rinsed with sterile water and allowed to
acclimatize to the medium for 7 days prior to NP exposure.
Three plants with uniform appearance were incubated in a
500-mL beaker containing 350 mL of the culture medium.
The beakers were placed in a thermostated chamber at
24±2 °C with 16 h light (180 μmol m−2 s−1) and 8 h dark
photoperiods. A stock suspension of ZnO at 100 mg L−1,
obtained by adding ZnO NPs to the culture medium, was
used to prepare ZnO suspension at different concentrations.
This stock suspension was treated by a sonicator (Vibra-
CellTM, USA; 50 Hz, 10 s pulse, and 5 s interval) for
10 min prior to use. Culture media with different ZnO
concentrations were prepared by adding the stock suspension
and then sonicated for 20 s. The plants were exposed to
different concentrations of ZnO NPs (0 (negative control),
1, 10, 20, and 50 mg L−1) for 7 days. A positive control,
which was exposed to ZnSO 4 at a concentration of
44 mg L−1 (corresponding to 10 mg L−1 Zn2+), was also
investigated. Each treatment was replicated five times.
After 7 days of exposure, the plants were harvested and
rinsed with distilled water. The surface water of the plants
was removed with filter papers, and their fresh weights
734
(FWs) were measured. The relative growth rate (RGR/day)
in each treatment was calculated using the formula:
RGR=(lnW2 −lnW1)/t. W1 and W2 are the initial and final
FW (gram), respectively, and t is the incubation time (day).
The dry weight (DW) of the plants was obtained after oven
drying at 80 °C for 48 h to constant weight.
Pigment assay
The contents of chl a, chl b, and carotenoid in the leaves of
the plants were analyzed according to Lichtenthaler (1987).
Freeze-dried leaves were cut into small pieces, and subsam-
ples of approximately 5 mg were extracted with 8 mL 96 %
ethanol in the dark at room temperature. After 24 h, the
pigment extracts were centrifuged at 5,000×g for 3 to
5 min after vigorous stirring. Then, absorbance was
measured at 665, 648, and 470 nm using a VIS-7220
spectrophotometer (Rayleigh Analytical Instrument
Corp., China).
Antioxidative enzyme assay
Superoxide dismutase (SOD) activity was determined using
the ferric cytochrome c method with xanthine/xanthine oxi-
dase as the source of superoxide radicals (McCord and
Fridovich 1969). Catalase (CAT) activity was assayed using
a U-3000 spectrophotometer (Hitachi, Tokyo, Japan) by
measuring the decrease in absorbance at 240 nm because of
H2O2 decomposition (Rao et al. 1996). Peroxidase (POD)
activity was determined using the methods described by Li
et al. (2000).
Aggregation of NPs and zinc concentration assay
Prior to the test, ZnO NPs were added to a 350-mL of culture
medium in a 500-mL beaker to obtain a final concentration
of 10 mg L−1. The suspension was treated by a sonicator as
previously described, and then, the beakers were placed in a
thermostated chamber at 24 ± 2 °C without disturbance.
Water samples (10 mL) were carefully obtained from the
upper layer of the suspension to avoid disturbance at each
hour of the initial 24 h to test for particle distribution. The
particle size of the ZnO NPs was determined using a
Nanotrac 250 particle analyzer (Microtrac Inc., USA).
The concentrations of zinc in the culture media and plants
of all treatments at 7 days were detected by inductively
coupled plasma-optical emission spectrometry (VISTA-
MPX, USA). The detailed methods for digestion were
according to our previous report (Hu et al. 2013). Briefly,
the plants were separated into leaves and roots, oven dried at
80 °C for 24 h, and then microwave digested first in 10 mL of
16 mM HNO3, followed by addition of 1 mL of H2O2. The
samples were diluted with 1 % (v/v) HNO3 to a final volume
Environ Sci Pollut Res (2014) 21:732–739
of 50 mL. The solution samples were filtered through
0.45 μm cellulose nitrate ultrafiltration membrane filters
(Whatman) and then acidified with HNO3 to a pH of
approximately 2.0.
Statistical analysis
All samples for growth assessment were replicated five
times, whereas other samples were replicated three times.
Three independent experiments were run. The results
presented are the arithmetic means with their corresponding
standard deviations. Differences between treatments were
tested for significance by ANOVA using Origin 7.0.
According to Tukey's multiple comparisons tests, *P<0.05
and **P<0.01 were considered significant and highly signifi-
cant, respectively.
Results
Characteristics of ZnO NPs
The results showed that the crystalline phase of ZnO
NPs was monocrystalline and that the mean size of the
single particle was approximately 25 nm (Fig. 1). BET
results showed that the surface area was approximately
90 m2 g−1.
Plant growth
Plant growth was evaluated as RGR and presented in
Fig. 2. No significant differences were observed
(P > 0.05) between all treatments with ZnO NPs and
the negative control. By contrast, the growth of S.
natans in the positive control (treated with ZnSO4)
was markedly inhibited (P<0.05).
Chlorophyll and carotenoid contents
Both chlorophyll and carotenoid were influenced by ZnO
NPs and ZnSO4 (Table 1). The contents of chl a and chl b
Fig. 1 Transmission electron microscopic image of ZnO nanoparticles
Environ Sci Pollut Res (2014) 21:732–739
Fig. 2 Relative growth rate (RGR) of S. natans during the 7-day
exposure
decreased with increasing ZnO NP dosage. The chl a content
of the plants at 50 mg L−1 ZnO NPs was approximately
2.57 mg g−1 DW (60 % of the control), which was
significantly lower than that of the negative control
(P<0.05). The chl a and chl b contents of the ZnSO4
treatment were significantly lower (P<0.05) than those
of the control and all ZnO NP treatments.
Carotenoid content slightly increased after treatment with
low concentration of ZnO NPs (1 mg L−1) and then de-
creased with increasing dosage. The carotenoid contents of
plants in the treatments with the highest concentration of
ZnO NPs and the positive control were significantly lower
(P<0.05) than that of the negative control.
Responses of antioxidant enzymes
Figure 3 shows the effects of the exposure on the activities of
SOD, CAT, and POD. SOD activity increased with increas-
ing NP dosages. Only at 50 mg L−1 ZnO NPs it showed
significant differences (P<0.05) from the negative control.
The SOD activity of S. natans exposed to 10 mg L−1 Zn2+
was significantly higher (P<0.01) than that of the control.
The response of CAT was similar to that of SOD. The
CAT activity of S. natans exposed to 50 mg L−1 ZnO NPs for
Table 1 Effects of ZnO nanoparticles and ZnSO4 on chl a, chl b, and carotenoid (in milligrams per gram DW) in the leaves of Salvinia natans
Parameters Control (0 mg L−1) Exposure groups
1 10
Chl a 4.06±0.45 4.15±0.33
Chl b 1.88±0.24 1.67±0.15
Carotenoid 0.69±0.08 0.74±0.10
*P<0.05, significant difference compared to the control
735
7 days was significantly higher (P<0.05) than that of the
control. The ZnSO4 treatment followed the trend of SOD.
POD activity initially increased with increasing ZnO NP
dosage and then markedly dropped (P<0.05) at a concentra-
tion of 50 mg L−1. The POD activity of S. natans exposed to
ZnSO4 for 7 days was significantly lower (P<0.01) than that
of the negative control.
Aggregation and dissolution of ZnO NPs
Figure 4 shows the particle size distribution (mean
values and standard deviations) of ZnO NPs in the
culture medium (without plants) during the first 24 h.
Although the suspension was initially treated with son-
ication, the mean particle size in the suspension imme-
diately after preparation was approximately 130 nm.
The particle size can reach more than 1.4 μm at 1 h,
suggesting rapid aggregation. After 8 h, the particle size
in the suspension reduced to approximately 1 nm. This
result indicated that no ZnO NPs were detected in the
supernatant. After treatment with ZnO NPs for 24 h,
white aggregates were observed at the bottom of the
beakers and the roots of the plants, especially for treat-
ments 20 and 50 mg L−1.
Table 2 shows the concentrations of zinc in the
culture media at 7 days. The medium of the negative
control allowed zinc to be one of the essential nutrients
at a concentration of 11.4 μg L−1, which could maintain
the growth of S. natans. The dissolution portions were
the principal sources of zinc in treatments 1, 10, 20,
and 50 mg L−1, with mean values of 0.41, 1.87, 2.57,
and 2.93 mg L−1, respectively. The concentration of
Zn2+ in the positive control was 8.71 mg L−1. Table 2
also provides the zinc content in the plants after 7 days
of exposure. S. natans accumulated more zinc when
Zn2+ concentration was increased in the culture medium.
No significant differences in zinc content between the
leaves and roots were observed in each treatment.
However, the zinc content of the unrinsed roots was
evidently higher (P<0.01) than that of the rinsed roots
in treatments 10, 20, and 50 mg L−1, indicating that
aggregates were adsorbed on the roots.
20 50 ZnSO4
3.98±0.28 3.75±0.41 2.57 ±0.36* 2.36±0.21*
1.74±0.23 1.62±0.25 1.45±0.22 1.16±0.16*
0.58±0.07 0.53±0.09 0.39±0.08* 0.45±0.11*
736
c decreased in the leaves of S. natans in this treatment,
Fig. 3 Activities of a SOD, b CAT, and c POD of S. natans after 7 days
of exposure to ZnO NPs and ZnSO4 with different concentrations
Discussion
The results in this study indicated that 50 mg L−1 ZnO NPs
can cause oxidative stresses to S. natans and depress the
photosynthetic pigments. However, plant growth was
not significantly affected. The adsorption of ZnO NPs
on the roots and the accumulation of Zn in the plants
were simultaneously noted.
FW was selected for the evaluation of plant growth
in this study. Using the mean value of several plants
for DW evaluation is difficult because of the large
Environ Sci Pollut Res (2014) 21:732–739
Fig. 4 Variation in particle sizes (mean±SD) of ZnO in the culture
medium (without plants) during the first 24 h
difference of the individual weight of S. natans. In
addition, measuring the DW of the same plant twice
(before and after exposure) is impractical. Therefore,
we merely tested the FW. The results showed that the
plants suffered no remarkable inhibition after treatment
with 50 mg L−1 ZnO NPs. However, SOD and CAT
activities increased and photosynthetic pigment contents
suggesting a stress from ZnO NPs. The results failed to
meet the growth response. The underlying reasons for
the failure may be attributed to the following. First, a
relative shorter duration was used in this study (7 days)
to evaluate the stress of ZnO NPs. Therefore, the bio-
chemical variation was not reflected in such a short
exposure period. Second, the individual plant of S.
natans is larger than organisms, such as microalgae,
duckweed, and Daphnia, which are generally used in
ecotoxicological testing, and then, growth was only
considered in limited reports. For instance, Mandal
et al. (2013) reported that the growth of S. natans that
was exposed to aluminum (240 μM) for 7 days was not
influenced, whereas MDA content significantly in-
creased (P <0.05). Sen and Bhattacharyya (1994) found
that both FW and DW of S. natans decreased with time
at 10, 20, and 60 μg mL−1 Ni(II) in the culture medi-
um after 1, 3, and 6 days of contact. In this study, ZnO
NPs with a concentration of 50 mg L−1 did not induce
the growth response of this plant. Therefore, antioxi-
dant enzymes and photosynthetic pigments are accept-
able indicators for S. natans and selected to evaluate
environmental stresses.
Antioxidant enzymes of living organisms can protect
cells against adverse effects of reactive oxygen species
(ROS). The enzymes SOD, CAT, and POD can control
cellular levels of ROS (Weckx and Clijsters 1996). The
Environ Sci Pollut Res (2014) 21:732–739
Table 2 Concentrations of Zn2+ in the culture media and Salvinia natans after 7 days of exposure
Exposure groups Culture medium (mg L−1) Leaves (mg g−1 DW)
0 0.011±0.002 0.015±0.006
1 0.41±0.08 0.45±0.13
10 1.87±0.32 2.61±0.72
20 2.57±0.19 3.17±0.56
50 2.93±0.29 3.65±0.81
ZnSO4 8.71±0.56 4.28±0.83
activities of these antioxidant enzymes maintain the
steady-state levels of ROS in cells. POD has a vital
function in scavenging these ROS. SOD and CAT func-
tion together to convert O2· and H2O2 into harmless
H2O and O2, reduce the formation of hydroxyl free
radical ·OH, and lower the overall free radical content
of cells. In the present study, SOD and CAT activities
increased in plants treated with 50 mg L−1 ZnO NPs
(Fig. 3a, b), indicating better ROS scavenging in the
system. Zinc was previously reported (Cakmak 2000;
Upadhyay and Panda 2010) to have a potential to in-
crease the biosynthesis of antioxidant enzymes in the
duckweed Spirodela polyrhiza. The activity of POD was
significantly inhibited by 50 mg L−1 ZnO NPs (Fig. 3c).
This result might be attributed to the stress from NPs or
Zn2+ that was beyond the protective ability of the enzymes.
Colloid is usually referred to particles in the 1-nm to
1-μm size range in aquatic systems. Therefore, ZnO
NPs initially existed in the suspension as colloid, aggre-
gated to large particles, and deposited. Colloidal fate
and behavior are dominated by aggregation (Buffle and
Leppard 1995; Gustafsson and Gschwend 1997), and
colloids ultimately aggregate to particles (>1 μm) and
deposit. In the present study, the aggregation of ZnO
NPs was significant in the culture medium, and substan-
tially, no ZnO NPs were observed in the supernatants
over 8 h (Fig. 4). This finding was basically consistent
with the previous reports (Adams et al. 2006; Franklin
et al. 2007), which evaluated the aggregation of ZnO
NPs in a freshwater system. We obtained samples from
the upper layer of the suspension because S. natans is a
free-floating fern and the variation of ZnO NPs in the
upper layer may affect the growth of the plants more
than those of other factors.
The dissolution rate of NPs is influenced by several
properties of NPs, including size, surface area, surface
curvature, and roughness (Borm et al. 2006). Moreover,
other factors, such as pH and ionic strength and aggre-
gation, may also significantly affect the dissolution. The
dissolution of ZnO is highly pH dependent. The pH
value of the culture medium used in this study was
6.5±0.2, and the Zn2+ concentration was 2.93 mg L−1
737
Roots (rinsed) (mg g−1 DW) Roots (unrinsed) (mg g−1 DW)
0.018±0.005 0.016±0.006
0.33±0.19 0.49±0.15
1.86±0.38 3.45±0.53
2.93±0.66 6.88±1.22
2.97±0.71 8.18±1.35
3.82±0.67 3.64±0.57
in treatment 50 mg L−1 ZnO NPs (Table 2), which was
close to a recent report (Wu et al. 2010). The soluble
form was regarded as the main bioavailable portion of
the total contaminant (Di Toro et al. 1991). Meanwhile,
Adams et al. (2006) and Franklin et al. (2007) also
reported the close relationship between the dissolved
portion and the toxicity of ZnO NPs. Both adsorption
of ZnO NPs on the roots and accumulation of zinc in
the plants were observed in our study. Significant dif-
ferences (P<0.01) in the measured concentration of zinc
were found between the rinsed and unrinsed roots.
Both aggregation and dissolution may influence the
stress of ZnO NPs to S. natans. However, which be-
havior of ZnO NPs contributed more to the stress is
not clearly elucidated. Stress of NPs depends on their
property, test organism species, and surrounding solu-
tion conditions (Lin and Xing 2008). According to
previous reports, aggregation of NPs reduced their tox-
icity on D. magna (Yu et al. 2011), and the toxicity of
ZnO NPs to zebrafish embryo might be due to the
combined effects of aggregation and dissolution (Zhu
et al. 2009). For plants, ZnO NPs with a diameter of
25 nm are hard to be directly taken up into the roots.
Asli and Neumann (2009) previously reported that
maize (Zea mays L.) roots exclude TiO2 NPs larger
than 6.6 nm after NP exposure, and inhibitory effects
of NP additions occurred on root hydraulic conductiv-
ity. The stress mechanism of ZnO NPs to S. natans
may be complex. Solubility could have an important
function. Meanwhile, the NPs covering the roots may also
influence the uptake of nutrients and water from the medium.
In summary, the present study investigated the adverse
effects of ZnO NPs on S. natans. The results showed that
50 mg L−1 NPs can adversely affect S. natans. ZnO NPs are
apt to aggregation and dissolution in the medium, and both
processes may affect their stress on the plant.
Acknowledgments This study was supported by the Shandong Out-
standing Young Scientist Award Fund (grant no. BS2010SF005), the
Open Fund from the Shandong Provincial Key Laboratory of Water and
Soil Conservation & Environmental Protection (grant no. stkf201203),
and the fund from Linyi University.
738
References
Adams LK, Lyon DY, Alvarez PJJ (2006) Comparative eco-toxicity of
nanoscale TiO2, SiO2, and ZnO water suspensions. Water Res
40:3527–3532
Aruoja V, Dubourguier HC, Kasemets K, Kahru A (2009) Toxicity
of nanoparticles of CuO, ZnO and TiO 2 to microalgae
Pseudokirchneriella subcapitata. Sci Total Environ 407:1461–1468
Asli S, Neumann PM (2009) Colloidal suspensions of clay or titanium
dioxide nanoparticles can inhibit leaf growth and transpiration via
physical effects on root water transport. Plant Cell Environ
32:577–584
Borm P, Klaessig FC, Landry TD, Moudgil B, Pauluhn J, Thomas K,
Trottier R, Wood S (2006) Research strategies for safety evalua-
tion of nanomaterials, part V: role of dissolution in biological fate
and effects of nanoscale particles. Toxicol Sci 90:23–32
Boxall A, Chaudhry Q, Sinclair C, Jones A, Aitken R, Jefferson B, Watts
C (2007) Current and future predicted environmental exposure to
engineered nanoparticles. Central Science Laboratory, York
Brayner R, Dahoumane SA, Yéprémian C, Djediat C, Ml M, Couté A,
Fiévet F (2010) ZnO nanoparticles: synthesis, characterization,
and ecotoxicological studies. Langmuir 26:6522–6528
Brunauer S, Emmett PH, Teller E (1938) Adsorption of gases in mul-
timolecular layers. J Am Chem Soc 60:309–319
Buffle J, Leppard GG (1995) Characterization of aquatic colloids and
macromolecules. 1. Structure and behavior of colloidal material.
Environ Sci Technol 29:2169–2175
Cakmak I (2000) Possible roles of zinc in protecting plant cells from
damage by reactive oxygen species. New Phytol 185–205
Dhir B, Sharmila P, Pardha Saradhi P, Nasim SA (2009) Physiological
and antioxidant responses of Salvinia natans exposed to
chromium-rich wastewater. Ecotoxicol Environ Saf 72:1790–1797
Di Toro DM, Zarba CS, Hansen DJ, Berry WJ, Swartz RC, Cowan CE,
Pavlou SP, Allen HE, Thomas NA, Paquin PR (1991) Technical
basis for establishing sediment quality criteria for nonionic organic
chemicals using equilibrium partitioning. Environ Toxicol Chem
10:1541–1583
Franklin NM, Rogers NJ, Apte SC, Batley GE, Gadd GE, Casey PS
(2007) Comparative toxicity of nanoparticulate ZnO, bulk ZnO,
and ZnCl 2 to a freshwater microalga (Pseudokirchneriella
subcapitata): the importance of particle solubility. Environ Sci
Technol 41:8484–8490
Gottschalk F, Sonderer T, Scholz RW, Nowack B (2009) Modeled
environmental concentrations of engineered nanomaterials (TiO2,
ZnO, Ag, CNT, fullerenes) for different regions. Environ Sci
Technol 43:9216–9222
Gubbins EJ, Batty LC, Lead JR (2011) Phytotoxicity of silver
nanoparticles to Lemna minor L. Environ Pollut 159:1551–1559
Gustafsson Ö, Gschwend PM (1997) Aquatic colloids: concepts, defi-
nitions, and current challenges. Limnol Oceanogr 42:519–528
Handy RD, Owen R, Valsami-Jones E (2008) The ecotoxicology of
nanoparticles and nanomaterials: current status, knowledge gaps,
challenges, and future needs. Ecotoxicology 17:315–325
Heinlaan M, Ivask A, Blinova I, Dubourguier HC, Kahru A (2008)
Toxicity of nanosized and bulk ZnO, CuO and TiO2 to bacteria
Vibrio fischeri and crustaceans Daphnia magna and
Thamnocephalus platyurus. Chemosphere 71:1308–1316
Hu C, Liu Y, Li X, Li M (2013) Biochemical responses of duckweed
(Spirodela polyrhiza) to zinc oxide nanoparticles. Arch Environ
Contam Toxicol 64:643–651
Jiang HS, Li M, Chang FY, Li W, Yin LY (2012) Physiological analysis
of silver nanoparticles and AgNO3 toxicity to Spirodela polyrhiza.
Environ Toxicol Chem 31:1880–1886
Kahru A, Dubourguier H-C (2010) From ecotoxicology to
nanoecotoxicology. Toxicology 269:105–119
Environ Sci Pollut Res (2014) 21:732–739
Kim E, Kim SH, Kim HC, Lee S, Lee S, Jeong S (2011) Growth
inhibition of aquatic plant caused by silver and titanium oxide
nanoparticles. Toxicol Environ Heal Sci 3:1–6
Lecoanet HF, Wiesner MR (2004) Velocity effects on fullerene and
oxide nanoparticle deposition in porous media. Environ Sci
Technol 38:4377–4382
Lecoanet HF, Bottero J-Y, Wiesner MR (2004) Laboratory assessment
of the mobility of nanomaterials in porous media. Environ Sci
Technol 38:5164–5169
Li H (2000) Principles and techniques of plant physiological biochemical
experiment. Higher Education Press, Beijing, pp 134–137 (in Chinese)
Lichtenthaler HK (1987) Chlorophylls and carotenoids: pigments of
photosynthetic biomembranes. In: Lester Packer RD (ed) Methods
in enzymology. Academic, London, pp 350–382
Lin D, Xing B (2008) Root uptake and phytotoxicity of ZnO
nanoparticles. Environ Sci Technol 42:5580–5585
Love SA, Maurer-Jones MA, Thompson JW, Lin Y-S, Haynes CL (2012)
Assessing nanoparticle toxicity. Annu Rev Anal Chem 5:181–205
Ma H, Williams PL, Diamond SA (2013) Ecotoxicity of manufactured
ZnO nanoparticles—a review. Environ Pollut 172:76–85
Mandal C, Ghosh N, Maiti S, Das K, Gupta S, Dey N, Adak MK (2013)
Antioxidative responses of Salvinia (Salvinia natans Linn.) to
aluminium stress and it's modulation by polyamine. Physiol Mol
Biol Plants 19:91–103
Masciangioli T, Zhang WX (2003) Peer reviewed: environmental tech-
nologies at the nanoscale. Environ Sci Technol 37:102A–108A
Maynard A, Michelson E (2006) The nanotechnology consumer prod-
ucts inventory. Woodrow Wilson International Center for
Scholars, Washington, DC
McCord JM, Fridovich I (1969) Superoxide dismutase: an enzymic
function for erythrocuprein (hemocuprein). J Biol Chem
244:6049–6055
Miller RJ, Lenihan HS, Muller EB, Tseng N, Hanna SK, Keller AA
(2010) Impacts of metal oxide nanoparticles on marine phyto-
plankton. Environ Sci Technol 44:7329–7334
Nel A, Xia T, Madler L, Li N (2006) Toxic potential of materials at the
nanolevel. Science 311:622–627
OECD (2006) Test No. 221: Lemna sp. growth inhabition test. OECD
guidelines for the testing of chemicals, section 2, OECD
publishing
Peralta-Videa JR, Zhao L, Lopez-Moreno ML, de la Rosa G, Hong J,
Gardea-Torresdey JL (2011) Nanomaterials and the environment:
a review for the biennium 2008–2010. J Hazard Mater 186:1–15
Petosa AR, Jaisi DP, Quevedo IR, Elimelech M, Tufenkji N (2010)
Aggregation and deposition of engineered nanomaterials in aquat-
ic environments: role of physicochemical interactions. Environ Sci
Technol 44:6532–6549
Rao MV, Paliyath G, Ormrod DP (1996) Ultraviolet-B- and ozone-
induced biochemical changes in antioxidant enzymes of
Arabidopsis thaliana. Plant Physiol 110:125–136
Sen A, Bhattacharyya M (1994) Studies of uptake and toxic effects of
NI(II) on Salvinia natans. Water Air Soil Pollut 78:141–152
Sharma VK (2009) Aggregation and toxicity of titanium dioxide
nanoparticles in aquatic environment—a review. J Environ Sci
Health A 44:1485–1495
Shi J, Abid AD, Kennedy IM, Hristova KR, Silk WK (2011) To
duckweeds (Landoltia punctata), nanoparticulate copper oxide is
more inhibitory than the soluble copper in the bulk solution.
Environ Pollut 159:1277–1282
Sun Q, Wang Q, Jena P, Kawazoe Y (2005) Clustering of Ti on a C60
surface and its effect on hydrogen storage. J Am Chem Soc
127:14582–14583
Upadhyay R, Panda SK (2010) Zinc reduces copper toxicity induced
oxidative stress by promoting antioxidant defense in freshly grown
aquatic duckweed Spirodela polyrhiza L. J Hazard Mater
175:1081–1084
Environ Sci Pollut Res (2014) 21:732–739
Weckx JEJ, Clijsters HMM (1996) Oxidative damage and defense mech-
anisms in primary leaves of Phaseolus vulgaris as a result of root
assimilation of toxic amounts of copper. Physiol Plant 96:506–512
Wiench K, Wohlleben W, Hisgen V, Radke K, Salinas E, Zok S, Landsiedel
R (2009) Acute and chronic effects of nano- and non-nano-scale TiO2
and ZnO particles on mobility and reproduction of the freshwater
invertebrate Daphnia magna. Chemosphere 76:1356–1365
Wu B, Wang Y, Lee YH, Horst A, Wang Z, Chen DR, Sureshkumar R,
Tang YJ (2010) Comparative eco-toxicities of nano-ZnO particles
under aquatic and aerosol exposure modes. Environ Sci Technol
44:1484–1489
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Yu LP, Fang T, Xiong DW, Zhu WT, Sima XF (2011) Comparative
toxicity of nano-ZnO and bulk ZnO suspensions to zebrafish and
the effects of sedimentation, ·OH production and particle dissolu-
tion in distilled water. J Environ Monitor 13:1975–1982
Zhu X, Zhu L, Duan Z, Qi R, Li Y, Lang Y (2008) Comparative toxicity
of several metal oxide nanoparticle aqueous suspensions to
zebrafish (Danio rerio) early developmental stage. J Environ Sci
Health A 43:278–284
Zhu X, Wang J, Zhang X, Chang Y, Chen Y (2009) The impact of ZnO
nanoparticle aggregates on the embryonic development of
zebrafish (Danio rerio). Nanotechnology 20:195103