Assessment of Effects of the Long-Term Exposure of Agricultural

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Assessment of effects of the long-term exposure of agricultural crops to carbon nanotubes Mohamed Hassen Lahiani, Zeid A Nima, Hector Villagarcia, Alexandru S. Biris, and Mariya Khodakovskaya J. Agric. Food Chem., Just Accepted Manuscript • DOI: 10.1021/acs.jafc.7b01863 • Publication Date (Web): 14 Aug 2017 Downloaded from http://pubs.acs.org on August 15, 2017

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Journal of Agricultural and Food Chemistry

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Assessment of Effects of the Long-term Exposure of Agricultural

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Crops to Carbon Nanotubes

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Mohamed H. Lahiani1, Zeid Nima2, Hector Villagarcia1, Alexandru S. Biris2, Mariya V.

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Khodakovskaya1*

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-Department of Biology, University of Arkansas at Little Rock, Little Rock, Arkansas, 72205.

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Center for Integrative Nanotechnology Science, University of Arkansas at Little Rock, Little

Rock , Arkansas, 72205. *Author for correspondence: [email protected]

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ABSTRACT

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Carbon-based nanomaterials (CBNs) were described as nanomaterials possessing abilities of

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plant growth regulators. Here, we investigated the effects of long-term exposure of multi-walled

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carbon nanotubes (MWCNTs) on the growth of three important crops (barley, soybean, corn).

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The cultivation of all tested species was carried in hydroponics supplemented with 50µg/ml of

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MWCNTs. After 20 weeks of continuous exposure to the nanomaterials, no significant toxic

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signs on plant development were observed. Several positive phenotypical changes were recorded

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in addition to the enhancement of photosynthesis in MWCNTs-exposed crops. Raman

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spectroscopy with point-by-point mapping proved that MWCNTs added to hydroponics solution

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moved into all tested species and were distributed in analyzed organs (leaves, stems, roots,

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seeds).

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agriculture. However, documented presence of MWCNTs in different organs of all exposed

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crops again highlighted the importance of detail risk assessment of nano-contaminated plants

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moving into the food chain.

Our results confirmed the significant potentials of CBN’s applications in plant

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KEYWORDS

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CBNs uptake, carbon nanotubes, plant growth, detection of carbon nanotubes in plants,

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hydroponics system

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INTRODUCTION

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A wide range of CBNs (carbon nanotubes, carbon nanohorns, graphene) may stimulate seed

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germination, plant growth 1-4, production of flowers/fruits 5 and activate plant cell division 1, 6. It

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was shown that CBNs are capable of penetrating plant cell walls as well as thick seed coat

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Positive effects of CBNs in planta are reproducible for different crop species

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achievable by a number of delivery methods including spray, introduction in soil/growth medium

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7, 10

1, 3, 7, 10-13

3, 7-9

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and

. Reproducibility and consistency of effects of CBNs are solid foundations for the

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development of new plant growth regulators based in “nano-formula”. However, in order to

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develop the technology that will be suitable for food crops and approved by Federal Agencies,

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many questions have to be answered. Firstly, it has to be proven that long-term exposure of

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plants to CBNs will not lead to toxic effects in planta. Previously, it had been demonstrated that

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positive effects of CBNs can be observed when CBNs were used in relatively low doses (10-100

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ug/ml) 7, 10, 14. However, long-term contact with plants with CBNs used as fertilizers may modify

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the response of exposed plants. Secondly, a comprehensive risk assessment of food crops

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exposed to CBNs to humans and animals should be performed at several levels including the

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confirmation of contamination of exposed plants with nanomaterials, toxicity tests using in vitro

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and in vivo experiments. The first group of risk assessment experiments (detection of

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nanomaterials on exposed plants) will rely on the development of sensitive and relatively simple

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analytical techniques. The detection of carbon nanomaterials in complex biological systems is

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rather difficult, given their carbon chemical structure, which cannot be easily discriminated from

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that of the actual biological system. The existing methods for detection and visualization of

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CBNs in organic tissues have many challenges. Using microscopic methods such as confocal

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laser scanning microscopy (CLSM)15, transmission electron microscopy (TEM) 16, and scanning 3 ACS Paragon Plus Environment


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electron microscopy (SEM)17,

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reason, these methods should be combined with other spectroscopic methods for confirmation of

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the nanoparticle presence. Two of the most promising spectroscopic techniques for the

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carbonaceous nanomaterials are Raman spectroscopy (based on light scattering) and

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photoacoustic microscopy (PAM) (based on light absorption). Raman spectroscopy uses the

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specific peaks characteristic to the CBNs and their intensity to detect and actually map the

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presence of these nanomaterials in the biological systems1, 7, 13, 19, 20. Therefore, the combination

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of CBNs exposure to plant systems and the Raman spectroscopy analysis of their presence and

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possibly distribution could elucidate some of the underlying phenomena at the interface between

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nanoscale materials and agriculturally valuable species.

, CNTs can be misled for plant cellular structures. For this

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The goal of this investigation was to understand the biological response of three crop species

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(barley, corn, and soybean) after long-term exposure to a representative type of CBNs (multi-

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walled carbon nanotubes, MWCNTs). The long-term exposure to MWCNTs was achieved by

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the cultivation of all tested species in hydroponics solution supplemented with MWCNTs for 20

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weeks. The response of plants on such treatment was monitored by phenotypical studies,

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characterization of the photosynthetic ability of exposed plants and proof of translocation of

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MWCNTs in different organs (Figure 1).

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MATERIALS AND METHODS Delivery of MWCNTs to plants though addition to hydroponic systems

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The hydroponic system was obtained from Hydrofarm ® (Grand Prairie, TX). Each tray

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contained six planters and plants were supported with clay pebbles. The system was linked with

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an air pump to keep roots and nutrient solutions well aerated. A water pump was used to provide 4 ACS Paragon Plus Environment

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a continuous flow of nutrients and carbon nanotubes in the solution. The trays were filled with

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10 L of deionized water and supplemented with 0.5 ml of nutrient solution per 1 L of water. The

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solution level was kept constant by checking the solution level daily, using a view/drain tube.

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Nutrient solutions were provided weekly for the hydroponic systems. The hydroponic control

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systems consisted of water and nutrients solution only. However, the treated hydroponic systems

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included a solution of MWCNT. MWCNTs were synthesized at the Center of integrative for

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Nanotechnology Sciences at the University of Arkansas at Little Rock. The Carbon

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nanomaterials were characterized as described earlier by Lahiani et al., (2013). MWCNTs were

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dispersed using a QSonica, LLC (Newtown, CT) sonicator. MWCNTs were added to the 2-week

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old seedlings in hydroponics for three consecutive weeks as following: 100 mg in week 1 and an

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addition of 200 mg in weeks 2 and 3. Using a water level indicator, the water level was

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maintained at 10L during the whole experiment. The growth of control plants and plant exposed

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to MWCNTs was monitored by: A) counting the number of leaves, internodes, fruits (soybean

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and corn), spikelet (barley) ; B) measuring the shoot length, internode length , fruit length, lateral

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branches (soybean and corn), tillers (barley); C) determining the total fresh/dry weight of shoots,

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leaves, roots and fruits.

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Statistics for tests of germination and plant growth All assays were performed in triplicate. All figures are represented as mean values ± SE

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(standard errors). All data were analyzed using SPSS® software by performing repeated measure

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ANOVA for time-effect analysis and ANOVA and posthoc analysis using the Tukey test for

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treatment differences. Statistical significance was determined by p Vicia faba L.) seedlings under combined stress of lead and cadmium. J.

386

Hazard. Mater. 2014, 274, 404-412.

387

33.

388

(< i> Amaranthus tricolor L) and the role of ascorbic acid as an antioxidant. J. Hazard.

389

Mater. 2012, 243, 212-222.

390

34.

391

(Amaranthus tricolor L) and the role of ascorbic acid as an antioxidant. J. Hazard. Mater. 2012,

392

243, 212-222.

Khodakovskaya, M. V.; Lahiani, M. H., Role of Nanoparticles for Delivery of Genetic

Begum, P.; Ikhtiari, R.; Fugetsu, B., Potential Impact of Multi-Walled Carbon Nanotubes

Begum, P.; Ikhtiari, R.; Fugetsu, B., Graphene phytotoxicity in the seedling stage of

Jiang, Y.; Hua, Z.; Zhao, Y.; Liu, Q.; Wang, F.; Zhang, Q. In The Effect of Carbon

Wang, C.; Liu, H.; Chen, J.; Tian, Y.; Shi, J.; Li, D.; Guo, C.; Ma, Q., Carboxylated

Begum, P.; Fugetsu, B., Phytotoxicity of multi-walled carbon nanotubes on red spinach

Begum, P.; Fugetsu, B., Phytotoxicity of multi-walled carbon nanotubes on red spinach


Page 19 of 37


393

35.

Giraldo, J. P.; Landry, M. P.; Kwak, S. Y.; Jain, R. M.; Wong, M. H.; Iverson, N. M.;

394

Ben‐Naim, M.; Strano, M. S., A Ratiometric Sensor Using Single Chirality Near‐Infrared

395

Fluorescent Carbon Nanotubes: Application to In Vivo Monitoring. Small 2015, 11, 3973–3984.

396

36. Kole, C.; Kole, P.; Randunu, K. M.; Choudhary, P.; Podila, R.; Ke, P. C.; Rao, A. M.;

397

Marcus, R. K., Nanobiotechnology can boost crop production and quality: first evidence from

398

increased plant biomass, fruit yield and phytomedicine content in bitter melon (Momordica

399

charantia). BMC biotechnology 2013, 13, 1.

400

37.

401

M.; Mizukami, H.; Bianco, A.; Baba, Y., Trafficking and subcellular localization of multiwalled

402

carbon nanotubes in plant cells. ACS nano 2011, 5, 493-499.

403

38.

404

plant cell biology: carbon nanotubes as organelle targeting nanocarriers. RSC Advances 2013, 3,

405

4856-4862.

406

39.

407

Limbach, L. K., No Evidence for Cerium Dioxide Nanoparticle Translocation in Maize Plants.

408

Environ. Sci. Technol. 2010, 44, 8718-8723.

409

40.

410

the brain of juvenile large mouth bass. Environ. Health Perspect 2004, 112, 1058-1062.

411

41.

412

Nanobiotechno 2014, 12, 16.

413

42.

414

Ono, A.; Kamata, E.; Hirose, A., No toxicological effects on acute and repeated oral gavage

Serag, M. F.; Kaji, N.; Gaillard, C.; Okamoto, Y.; Terasaka, K.; Jabasini, M.; Tokeshi,

Serag, M. F.; Kaji, N.; Habuchi, S.; Bianco, A.; Baba, Y., Nanobiotechnology meets

Birbaum, K.; Brogioli, R.; Schellenberg, M.; Martinoia, E.; Stark, W. J.; Günther, D.;

Oberdorster, E., Manufactured nanomaterials (fullerenes, C60) induce oxidative stress in

Husen, A.; Siddiqi, K. S., Carbon and fullerene nanomaterials in plant system. J

Matsumoto, M.; Serizawa, H.; Sunaga, M.; Kato, H.; Takahashi, M.; Hirata-Koizumi, M.;



415

doses of single-wall or multi-wall carbon nanotube in rats. The Journal of toxicological sciences

416

2012, 37, 463-474.

417

43.

418

damaged DNA in rats exposed by oral gavage to C60 fullerenes and single-walled carbon

419

nanotubes. Environmental health perspectives 2009, 117, 703–708.

420

44.

421

nanotubes in biological samples through microwave-induced heating. Carbon 2012, 50, 4441-

422

4449.

423

45.

424

F.; Carriere, M., Accumulation, translocation and impact of TiO2 nanoparticles in wheat

425

(Triticum aestivum spp.): Influence of diameter and crystal phase. Sci. Total Environ. 2012, 431,

426

197-208.

427

46.Long, S.P.; Zhu, X.; Naidu,S. L.;Ort, D.R.,Can improvement in photosynthesis increase crop

428

yield. Plant, Cell and Environment (2006) 29, 315-330

429

47. Evans, J.R., Improving photosynthesis. Plant Physiol. 2013, 162:1780-1793

Page 20 of 37

Folkmann, J. K.; Risom, L.; Jacobsen, N. R.; Wallin, H.; Loft, S.; Moller, P., Oxidatively

Irin, F.; Shrestha, B.; Cañas, J. E.; Saed, M. A.; Green, M. J., Detection of carbon

Larue, C.; Laurette, J.; Herlin-Boime, N.; Khodja, H.; Fayard, B.; Flank, A.-M.; Brisset,

430 431 432 433 434 435 436 437 20 ACS Paragon Plus Environment

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Figure Captions

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Figure 1. Experiment design focused of monitoring effects of long-term exposure of crop

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species (barley, corn, and soybean) to MWCNT through cultivation in hydroponics system

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supplemented with MWCNT. Experimental stages involved the preparation of hydroponics

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solution, cultivation stage (20 weeks), and the bio-effects monitoring stage.

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Figure 2. Average Photosynthetic light-response curves for MWCNTs treated and non-treated

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corn (A) and soybean (B) plants growing in hydroponic systems supplemented with MWCNT.

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These curves are the average of four plant measurement. All the measurements were conducted

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at 400 ppm CO2 and ~25°C. *, p

Assessment of Effects of the Long-Term Exposure of Agricultural

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