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Impact of copper oxide nanoparticles exposure on Arabidopsis thaliana growth, root system development, root lignificaion, and molecular level changes

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Abstract

The effect of copper oxide nanoparticles (CuONPs) on physiological and molecular level responses were studied in Arabidopsis thaliana. The seedlings were exposed to different concentrations of CuONPs (0, 0.5, 1, 2, 5, 10, 20, 50, and 100 mg/L) for 21 days in half strength Murashige and Skoog medium. The plant biomass significantly reduced under different concentrations (2, 5, 10, 20, 50, and 100 mg/L) of CuONPs stress. Exposure to 2, 5, 10, 20, 50, and 100 mg/L of CuONPs has resulted in significant reduction of total chlorophyll content. The anthocyanin content significantly increased upon exposure to 10, 20, 50, and 100 mg/L of CuONPs. Increased lipid peroxidation was observed upon exposure to 5, 10, and 20 mg/L of CuONPs and amino acid proline content was significantly high in plants exposed to 10 and 20 mg/L of CuONPs. Significant reduction in root elongation was observed upon exposure to 0.5–100 mg/L of CuONPs for 21 days. Exposure to CuONPs has resulted in retardation of primary root growth, enhanced lateral root formation, and also resulted in loss of root gravitropism. Staining with phloroglucionol detected the deposition of lignin in CuONPs-treated roots. Histochemical staining of leaves and roots of CuONPs-exposed plants with nitroblue tetrazolium and 3′3′-diaminobenzidine showed a concentration-dependant increase in superoxide and hydrogen peroxide formation in leaves and roots of CuONPs-exposed plants. Cytotoxicity was observed in root tips of CuONPs-exposed plants as evidenced by increased propidium iodide staining. Real-time PCR analysis showed significant induction of genes related to oxidative stress responses, sulfur assimilation, glutathione, and proline biosynthesis under CuONPs stress.

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Acknowledgments

This paper was supported by the SMART-Research Professor Program of Konkuk University, Seoul, South Korea to Dr. Prakash M. Gopalakrishnan Nair. This work was supported by a grant from the Next-Generation BioGreen 21 Program (Plant Molecular Center No. PJ009053), Rural Development Administration, Republic of Korea.

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Authors and Affiliations

  1. Department of Applied Biosciences, College of Life and Environmental Sciences, Konkuk University, Seoul, South Korea

    Prakash M. Gopalakrishnan Nair & Ill Min Chung

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  1. Prakash M. Gopalakrishnan Nair
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  2. Ill Min Chung
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Correspondence to Ill Min Chung.

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Responsible editor: Elena Maestri

Highlights

• Studied the physiological and molecular level responses of CuONPs stress in A. thaliana.

• CuONPs stress significantly reduced plant biomass and root growth.

• CuONPs stress caused root growth modifications and enhanced lateral root formation.

• Histochemical staining revealed excess ROS generation.

• Propidium iodide staining indicated cell death in root apex.

• Induced antioxidant, sulfur assimilation, GSH biosynthesis genes.

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Supplementary Figure 1

Characterization of copper oxide nanoparticles using transmission electron microscopy (A, B) Morphology of CuONPs in deionized water, (C) Graph showing size distribution of copper oxide nanoparticles in deionized water based on dynamic light scattering analysis and (D, E) morphology of copper oxide nanoparticles in ½ MS medium. (GIF 735 kb)

High resolution image (TIFF 2227 kb)

Supplementary Figure 2

Phenotypes of A. thaliana plants grown in the presence of Cu2+ ions for 21 days (left to right: Control, 0.1 and 0.2 mg/L of Cu2+ ions) (B, C) Plants showing changes in root morphology and (D, E) loss of root gravitropism after exposure to 10 and 20 mg/L of copper oxide nanoparticles for 21 d. (GIF 1215 kb)

High resolution image (TIFF 3021 kb)

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Nair, P.M.G., Chung, I.M. Impact of copper oxide nanoparticles exposure on Arabidopsis thaliana growth, root system development, root lignificaion, and molecular level changes. Environ Sci Pollut Res 21, 12709–12722 (2014). https://doi.org/10.1007/s11356-014-3210-3

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  • DOI: https://doi.org/10.1007/s11356-014-3210-3

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