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EFFECTS OF NANO OXIDES ON HELIANTHUS ANNUUS IN A

CONTAMINATED SOIL
Domingo Martínez-Fernández, M. Vítková, M. Komárek
Czech University of Life Sciences Prague, Faculty of Environmental Sciences, Department of Environmental Geosciences, Kamycka 129, 165 21, Prague 6, Czech Republic

Email: domingo.marfer@gmail.com; martinez-fernandez@fzp.czu.cz

HIGHLIGHTS
• Nanomaghemite (NM) promoted the growth of H. annuus in a contaminated soil, mainly due to the immobilization of available Zn in the soil, with the
consequent reduction of its uptake and toxicity to the roots and aerial parts.
• During drought, NM looks not to interfere in plant physiological responses since no differences were detected in water-stress indicators.
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1 INTRODUCTION
The iron oxide nanomaterials can decrease trace elements (TEs) bioavailability in
contaminated soil by their huge reactivity and relatively-large specific surface area and
excellent sorption properties [1].
The scarce knowledge about mechanisms by which nanoparticles penetrate plants is
hindering their full application. We do not have a good understanding of their uptake, and
more studies need to be done at the root–soil interface.
This experiment studies the effects the NM on the plant physiology by the changes in
the availability of TEs for plants, focusing in plant–water relations during drought.

2 EXPERIMENT DESIGN
• H. annuus in pots with contaminated soil from the mining area of La Unión-Cartagena (Murcia, Spain) [2].
• Treatments: (i) control without any treatment; (ii) application of compost; and (iii) combined treatment with compost+NM.
• Dose of compost to increase a 1% the OM. Dose of 1% of NM mixed with the soil for optimal removal rate of TEs [1].
• Nanomaghemite (NM) as Fe (III) (γ-Fe2O3; 20-100 nm of particle size; Sigma Aldrich (Germany)).
• Soil maintained at 80% WHC during 42 days reduced to 30% WHC in order to impose water stress to the plants.
• Soil pore water samples were obtained using rhizon-samplers and analysed by ICP-OES.
• When the plants manifested symptoms caused by the drought (50d), they were harvested for biomass and TEs analysis.
• Plant water status evaluated by leaf relative water content (RWC) and Proline and total amino acids concentrations, as
indicators of stress in the plants during the drought-NM interaction [3, 4].

pH

Cd
µg g-1

Cu
µg g-1

Fe
mg g-1

Soil La Unión

7.49

31

142

153

5.15

Compost

8.80

-

48

1.6

Cd

Cu

Fe

µg L-1

µg L-1

µg L-1

3 ADSORPTION OF TEs BY NM
Although the use of compost is a suitable way to
improve soil structure and provide nutrients and OM, it
can produce an initial mobilization of TEs, as observed here
for Mg, Mn and also for Na in the pore water. However, the
combined use of compost+NM did not cause a significant
mobilization of these elements probably by their
adsorption onto the NM.
The immobilization of Mn, Mg and Na by NM may have
been helped by the supply of K+ by the compost, since
increasing concentrations of K+ can increase the adsorption
of Mg, Mn and Na onto NM in the soil solution [5].
A significant decrease of the available Zn in the pore
water was only observed when the compost was combined
with NM, presumably due to the sum of their adsorption
capacities.
Although some nanoparticles can be found in the plant’s
cells and tissues [6], no uptake or toxicity have been
reported for nano oxides, may be by the adherence of the
nanoparticles to the soil particles, and since the cell wall
pore sizes vary from 2–20 nm [6].

Pore water

pH

K
Mg
mg g-1 µg g-1

Mn
µg g-1

P
µg g-1

Pb
µg g-1

Zn
µg g-1

16567

4.07

696

6130

12506

43.4

7954

0.09

4950

36

141

K

Mg

Mn

Na

Pb

Zn

µg L-1

mg L-1

µg L-1

µg L-1

mg L-1 mg L-1

Control

7.44

1.4

77.4

9.0

9b

97 b

712 b

405 b

4.6

470 a

Compost

7.46

8.6

89.6

4.5

309 a

121 a

873 a

644 a

13.0

429 ab

Compost+NM

7.48

0.8

78.2

3.6

263 a 108 ab 780 ab 542 ab

6.6

366 b

ns

ns

ns

ns

***

*

*

**

ns

*

Cd

Cu

Fe

K

Mg

Mn

Na

ANOVA
Tissue

(µg L-1)

Pb

Zn

(µg L-1) (µg L-1) (µg L-1) (µg L-1) (µg L-1) (µg L-1) (µg L-1) (µg L-1)

AAs

(µmol g-1) (µmol g-1)

Shoot 1.92 ab 4.72 ab

50

6.68 b

0.87 a

0.32

202 a

67.3

0.381

8.39

Compost

Shoot 2.04 a

5.33 a

48

10.53 a 66 ab 2158 b 0.76 ab

0.80

178 a

61.4

0.407

9.55

Compost+NM

Shoot 1.60 b

4.66 b

52

9.44 a

49 b

0.65

127 b

61.1

0.59

9.08

*

*

ns

***

*

ns

***

ns

ns

ns

7038

10.15 a 444 a 1158 a

-

0.61

10.46

8.65 a 2822 b 213 ab 7.62 b 297 ab 894 ab

-

0.47

9.75

-

0.61

10.25

ns

ns

Control

Root

9.85

20.40 a

Compost

Root

8.87

16.29 ab 3988

Compost+NM

Root

6.89

14.94 b

ns

*

ANOVA

3587 a

(%)

Proline

Control

ANOVA

73 a

RWC

2013 b 0.62 b
***

3.83 b 4622 a 334 a

4813 7.48 ab 2540 b 181 b
ns

***

***

*

**

6.99 b

247 b

702 b

**

*

**

4 NM AS AMENDMENT DURING PHYTOREMEDIATION



The supply of OM by the compost could explain a stimulation of growth, but the fact that only the treatment with compost+NM
was able to produce a higher biomass suggests that this stimulation was caused by the NM and their role in the immobilization of
TEs in the soil fraction.
Proline accumulation is a common metabolic response in higher plants during water deficit, used as index for
stress [7]. The presence of NM in the soil did not impact on the response of H. annuus to the water stress, since
plants with compost+NM had the same RWC, proline and total AAs content as the control and compost ones.
This results highlight the applicability of NM during phytostabilization in contaminated soils, by the
immobilization of the TEs in the soil and allowing the growth of plants where the contaminants are less available.
Further research is needed to clarify whether such root functionality is affected by NPs in other types of soils, as
well as in hydroponic cultures where their direct effect on the surface of roots can be investigated.

REFERENCES: [1] Komárek et al., 2013. Chemical stabilization of metals and arsenic in contaminated soils using oxides - A review. Environ. Pollut. 172: 9-22. [2] Martínez-Fernández et al., 2014. Comparison of compost and humic
fertiliser effects on growth and trace elements accumulation of native plant species in a mine soil phytoremediation experiment. Ecological Engineering 73: 588-597. [3] Bates et al., 1973. Rapid determination of free proline for water
stress studies. Plant Soil 39: 205-207. [4] Lee and Takahashi 1966. An improved colorimetric determination of amino acids with the use of ninhydrin. Analytical Biochemistry 14: 71-77. [5] Martínez-Fernández et al., 2014. Trace
elements and nutrients adsorption onto nano-maghemite in a contaminated-soil solution: A geochemical/statistical approach. Journal of Hazardous Material 276: 271-277. [6] Rico et al., 2011. Interaction of nanoparticles with edible
plants and their possible implications in the food chain. Journal of Agricultural and Food Chemistry 59: 3485-3498. [7] Verbruggen and Hermans. 2008. Proline accumulation in plants: a review. Amino Acids 35: 753-759.
ACKNOWLEDGEMENTS
The authors thank Sylva Číhalová and the PhytoRec Team of CEBAS-CSIC their assistance with analyses., and to the European project Postdok ČZU (ESF/MŠMT CZ.1.07/2.3.00/30.0040) and the Czech Science Foundation (GAČR 503/11/0840) for financial support .