Professional Documents
Culture Documents
Tmp522e TMP
Tmp522e TMP
Environmental Science and Engineering PhD program, The University of Texas at El Paso, 500 West University Ave., El Paso, TX 79968, USA
Department of Chemistry, The University of Texas at El Paso, 500 West University Ave., El Paso, TX 79968, USA
University of California Center for Environmental Implications of Nanotechnology (UC CEIN), USA
d
Foundation Institute of Materials Science, IWT, University of Bremen, Badgasteiner Str. 3, 28359 Bremen, Germany
b
c
h i g h l i g h t s
g r a p h i c a l a b s t r a c t
a r t i c l e
i n f o
Article history:
Received 8 May 2014
Received in revised form 26 June 2014
Accepted 28 June 2014
Available online 8 July 2014
Keywords:
Nanoparticles
Iron doped zinc oxide nanoparticles
Phytotoxicity
Catalase
Ascorbate peroxidase
a b s t r a c t
Iron doping has shown to reduce toxicity of zinc oxide nanoparticles (ZnO NPs) in several organisms. To the
best of authors knowledge, this is the rst report on toxicological studies of Fe@ZnO NPs on terrestrial plants.
In this study, green pea plants (Pisum sativum L.) were grown for 25 days in soil treated with 10% Fe@ZnO NPs
at 0500 mg/kg. Effects were compared with our previous study where phytotoxicity of bare-ZnO NPs had
been investigated on green pea plants grown under similar environmental conditions. Different physiological and biochemical growth parameters were measured. Results showed increased Zn bioaccumulation in
roots (200%) and stems (3148%) as the exposed NP concentration increased (p 6 0.05) but Fe absorption
was not affected. At 500 mg/kg Fe@ZnO NPs treatment, chlorophyll content (27%) and H2O2 production
(50%) decreased signicantly (p 0.05) compared to control. Toxicity of doped ZnO NPs is less than that
of bare ZnO NPs as per zinc uptake, chlorophyll content, and ROS (H2O2) production are considered. Therefore, iron doping can be considered as a safer approach to reduce toxicity of ZnO NPs in terrestrial plants.
2014 Elsevier B.V. All rights reserved.
1. Introduction
Nanomaterials (NMs) possess at least one dimension less than
100 nm [1,2] and have enormous surface to volume ratio that is
Corresponding author at: Department of Chemistry, The University of Texas at El Paso, 500 West University Ave., El Paso, TX 79968, USA. Tel.: +1 915 747 5359;
fax: +1 915 747 5748.
E-mail address: jgardea@utep.edu (J.L. Gardea-Torresdey).
http://dx.doi.org/10.1016/j.cej.2014.06.112
1385-8947/ 2014 Elsevier B.V. All rights reserved.
395
396
Fig. 1. Rietveld renement of pure and Fe@ZnO for the extraction of cell parameters and crystallite sizes. The cell parameters of ZnO: wurtzite structure, a = b = 3.2477,
c = 5.2043, V = 47.54 3; Fe@ZnO: wurtzite structure, a = b = 3.2546, c = 5.2174, V = 47.86 3.
397
Fig. 2. (a) and (d): Low resolution images showing particle morphology (b) and (e) single particle HRTEM images (c) and (f) selected area diffraction patterns of pure and 10%
Fe doped ZnO NPs respectively. The results show that the doping is homogeneous and single crystalline with no detectable changes in the lattice spacings.
Table 1
Particle size evaluation of pure and 10% Fe doped ZnO NPs.
Sample
BET (dBET)
(nm)
XRD (dXRD)
(nm)
TEM (dTEM)
(nm)
Size in water
(nm)
Pure ZnO
10% Fe-ZnO
20.2
8.3
15.8
5.5
22
10
130
1780
Fig. 3. Root and stem lengths (cm) of green pea plants grown for 25 days in soil,
treated with 0(control)-500 mg/kg Fe@ZnO NPs. Bars represent mean of four
replicates standard error. Bars with the same letters show no statistically
signicant difference at p 6 0.05.
398
Fig. 4. Chlorophyll contents (SPAD) in the leaves of green pea plants grown for
25 days in soil, treated with 0(control)-500 mg/kg Fe@ZnO NPs. Bars represent
mean of four replicates standard error. Bars with the same letters show no
statistically signicant difference at p 6 0.05.
Fig. 5. (A) Zn and (B) Fe concentrations in roots, stems, and leaves in green peas grown for 25 days in soil, treated with 0(control)-500 mg/kg Fe@ZnO NPs. Bars represent
mean of four replicates standard error. Bars with the same letters show no statistically signicant difference at p 6 0.05. Comparisons were made between same tissues of
different treatments.
399
Table 2
Bioaccumulation factors under different treatments.
Bioaccumulation factors
125 mg/kg
250 mg/kg
500 mg/kg
Root
Stem
Leaf
Root
Stem
Leaf
Root
Stem
Leaf
1.8
0.44
0.28
0.74
0.24
0.15
0.72
0.15
0.09
Fig. 6. H2O2 concentration in roots, stems, and leaves of green pea plants grown for
25 days in soil treated with 0(control)-500 mg/kg Fe@ZnO NPs. Bars represent mean
of four replicates standard error. Bars with same letters shows statistically
signicant difference at p 6 0.05. Comparisons were made between same tissues of
different treatments.
p 6 0.05). In roots, there were 73%, 49%, and 33% decrease in APOX
activity with 125, 250, and 500 mg/kg treatments, respectively,
compared to control. In case of stem, a similar trend was observed.
The reduction was 84%, 86%, and 74% at 125, 250, and 500 mg/kg
treatments, respectively, compared to control. Mukherjee et al.
reported similar trend, where bare ZnO NP decreased APOX activity in root and leaf of green pea plants [26]. This nature of the curve
can also be correlated from a previous study performed by Hernandez-Viezcas et al. in velvet mesquite (Prosopis juliora-velutina),
where the authors reported a similar trend in APOX activity with
increasing ZnO NP concentration [41].
In roots and leaves, CAT activity was signicantly increased at
500 mg/kg treatment compared to all other treatments (Fig. 7B).
However, in stem, Fe@ZnO NPs did not affect the CAT activity.
Green pea leaves showed elevated level of CAT activity at 250
and 500 mg/kg treatments, compared to control. These results
are in accordance with the study performed by Hernandez-Viezcas
et al. with velvet mesquite (P. juliora-velutina), where increase in
CAT activity only at the highest concentration (4000 mg/kg) [41].
However, decreased CAT activity was reported in green pea leaves
with increasing bare ZnO NP doses, signifying higher ROS activity
in leaves [26]. The differential behavior of bare and doped ZnO
NPs may be attributed towards different size, surface charge, and
dissolution properties of the NPs [24]. Abiotic stress has shown
to decrease APOX activity, which lowers the H2O2 scavenging efciency inside the plant tissue [42]. This in turn may increase the
ROS level, which may surpass the ROS scavenging capacity of
CAT [43], resulting in an increase in CAT activity.
400
Acknowledgements
This material is based upon work supported by the National
Science Foundation and the Environmental Protection Agency
under Cooperative Agreement Number DBI-0830117. Any opinions, ndings, and conclusions or recommendations expressed in
this material are those of the author(s) and do not necessarily
reect the views of the National Science Foundation or the Environmental Protection Agency. This work has not been subjected
to EPA review and no ofcial endorsement should be inferred.
The authors also acknowledge the USDA Grant number 201138422-30835 and the NSF Grant # CHE-0840525. J.L. Gardea-Torresdey acknowledges the Dudley family for the Endowed Research
Professorship in Chemistry.
References
Fig. 7. (A) APOX and (B) CAT concentrations in roots, stems, and leaves of green pea
plants grown for 25 days in soil, treated with 0(control)-500 mg/kg Fe@ZnO NPs.
Bars represent mean of four replicates standard error. Bars with the same letters/
symbols show no statistically signicant difference at p 6 0.05. Comparisons were
made between same tissues of different treatments.
4. Conclusions
These ndings conrm that Fe@ZnO NPs did not produce
visible signs of toxicity including necrosis, stunting, chlorosis or
wilting in all the treatments. However, Fe@ZnO NPs affected different physiological and biochemical parameters in terms of plant
growth, chlorophyll content, ROS (H2O2) production, and antioxidative enzyme activity. Nevertheless, as per relative chlorophyll
content and ROS production are concerned, we found that
Fe@ZnO is less toxic than ZnO NPs in green peas grown under
the same conditions. Key ndings of this work support the fact
that greater hydrodynamic diameter and/or lesser dissolution
from Fe@ZnO NPs make it less available for green pea plants,
and hence lesser phytotoxicity compare to that of bare ZnO
NPs. Therefore, iron doping may be considered as an effective
alternative approach to reduce the toxicity of ZnO NPs in higher
terrestrial plants. Future studies have to be performed in wider
range of NP concentrations to determine the mechanisms of
toxicity and the effects on fruit quality.
401
[32] M.M.A. Bradford, A rapid and sensitive method for the quantitation of
microgram quantities of protein utilizing the principle of protein-dye
binding, Anal. Biochem. 72 (1976) 248254.
[33] H. Aebi, Catalases, in: H.U. Bergmeyer (Ed.), Methods of Enzymatic Analysis,
vol. 2, Academic Press, New York, 1974, pp. 673684.
[34] Y. Nakano, K. Asada, Hydrogen peroxide is scavenged by ascorbate-specic
peroxidase in spinach chloroplasts, Plant Cell Physiol. 22 (1981) 867880.
[35] J. Albertsson, S.C. Abrahams, A. Kvick, Atomic displacement, anharmonic
thermal vibration, expansivity and pyroelectric coefcient thermal
dependences in ZnO, Acta Crystallogr. Sect. B: Struct. Sci. 45 (1) (1989) 3440.
[36] S. Pokhrel, J. Birkenstock, M. Schowalter, A. Rosenauer, L. Mdler, Growth of
ultrane single crystalline WO3 nanoparticles using ame spray pyrolysis,
Cryst. Growth Des. 10 (2) (2010) 632639.
[37] D. Lin, B. Xing, Phytotoxicity of nanoparticles: inhibition of seed germination
and root growth, Environ. Pollut. 150 (2007) 243250.
[38] W.-M. Lee, Y.-J. An, H. Yoon, H.-S. Kweon, Toxicity and bioavailability of copper
nanoparticles to the terrestrial plants mung bean (Phaseolus radiatus) and
wheat (Triticum aestivum): plant agar test for water-insoluble nanoparticles,
Environ. Toxicol. Chem. 27 (9) (2008) 19151921.
[39] C.H. Foyer, S. Shigeoka, Understanding oxidative stress and antioxidant
functions to enhance photosynthesis, Plant Physiol. 155 (1) (2011) 93100.
[40] G.R. Rout, P. Das, Effect of metal toxicity on plant growth and metabolism: I.
Zinc, Agronomie 23 (2003) 311.
[41] J.A. Hernandez-Viezcas, H. Castillo-Michel, A.D. Servin, J.R. Peralta-Videa, J.L.
Gardea-Torresdey, Spectroscopic verication of zinc absorption and
distribution in the desert plant Prosopis juliora-velutina (velvet mesquite)
treated with ZnO nanoparticles, Chem. Eng. J. 170 (13) (2011) 346352.
[42] S. Komatsu, A.H.M. Kamal, T. Makino, Z. Hossain, Ultraweak photon emission
and proteomics analyses in soybean under abiotic stress, BBA-Proteins
Proteom. 1844 (7) (2014) 12081218.
[43] T. Xia, M. Kovochich, J. Brant, M. Hotze, J. Sempf, T. Oberley, C. Sioutas, J.I. Yeh,
M.R. Wiesner, A.E. Nel, Comparison of the abilities of ambient and
manufactured nanoparticles to induce cellular toxicity according to an
oxidative stress paradigm, Nano Lett. 6 (2006) 17941807.