Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 135, Issue 2, pp 223–234 | Cite as

Toxicity impacts of chemically and biologically synthesized CuO nanoparticles on cell suspension cultures of Nicotiana tabacum

  • Sepideh Mahjouri
  • Ali Movafeghi
  • Baharak Divband
  • Morteza Kosari-Nasab
Original Article


Nanotechnology has quite a lot of applications in various fields of industrial sectors like food and agriculture. Although nanotechnology can improve the quality of life, its possible associated risks should be assessed. Here copper oxide nanoparticles (CuO NPs) were synthesized by chemical (polymer pyrolysis) and biological (green) methods with an average size of 30 and 44 nm, respectively. Afterwards, a cell biology approach was applied to evaluate the toxic effects of chemically and biologically synthesized CuO nanoparticles on tobacco cell suspension cultures. Both types of CuO nanoparticles significantly dropped the viability of the cells in a dose and time dependent manner. Accordingly, tobacco cells were found to increase the activity of antioxidant enzymes after 48 h of exposure to nanoparticles. The production of reactive oxygen species (ROS) and malondialdehyde (MDA) in a dose dependent manner was also observed. Assessment of the toxicity of CuO NPs revealed that chemically synthesized NPs were more toxic than biologically synthesized ones. It can be concluded that the organic components of the plant extract as capping agents that remain on the surface of green synthesized CuO NPs may reduce their toxicity effects.


Nicotiana tabacum Cell culture Cytotoxicity CuO nanoparticles Green synthesis 



Chemically synthesized copper oxide nanoparticles


Green synthesized copper oxide nanoparticles




Reactive Oxygen Species


Superoxide dismutase







The authors thank the University of Tabriz (Iran) and the Hayyan Biotechnology Laboratory for scientific and financial supports.

Author contributions

AM and SM was responsible for the design and overall investigation. BD helped in synthesis and analysis of nanoparticles. SM and MK-N conducted the cell culture experiments. All authors read and approved the final manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Acharyulu N, Dubey R, Swaminadham V, Kollu P, Kalyani R, Pammi S (2014) Green synthesis of CuO nanoparticles using Phyllanthus amarus leaf extract and their antibacterial activity against multidrug resistance bacteria. Int J Engin Res Technol 3:639–641Google Scholar
  2. Adhikari T, Kundu S, Biswas AK, Tarafdar JC, Rao AS (2012) Effect of copper oxide nanoparticle on seed germination of selected crops. J Agric Sci Technol 2:815–823Google Scholar
  3. Azam A, Ahmed AS, Oves M, Khan M, Memic A (2012) Size-dependent antimicrobial properties of CuO nanoparticles against Gram-positive and-negative bacterial strains. Int J Nanomedicine 7:3527–3535. CrossRefPubMedPubMedCentralGoogle Scholar
  4. Boominathan R, Doran PM (2002) Ni-induced oxidative stress in roots of the Ni hyperaccumulator, Alyssum bertolonii. New Phytol 156:205–215. CrossRefGoogle Scholar
  5. Borm PJ et al (2006) The potential risks of nanomaterials: a review carried out for ECETOC. Part Fibre Toxicol 3:1743–8977. CrossRefGoogle Scholar
  6. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. ‎Anal Biochem 72:248–254. CrossRefPubMedGoogle Scholar
  7. Chance B, Maehly A (1955) [136] Assay of catalases and peroxidases. Methods Enzymol 2:764–775. CrossRefGoogle Scholar
  8. Choochote W, Suklampoo L, Ochaikul D (2014) Evaluation of antioxidant capacities of green microalgae. J Appl Phycol 26:43–48. CrossRefGoogle Scholar
  9. Comotto M, Casazza AA, Aliakbarian B, Caratto V, Ferretti M, Perego P (2014) Influence of TiO2 nanoparticles on growth and phenolic compounds production in photosynthetic microorganisms. Sci World J 2014:9. CrossRefGoogle Scholar
  10. Dimkpa CO et al (2012) CuO and ZnO nanoparticles: phytotoxicity, metal speciation, and induction of oxidative stress in sand-grown wheat. J Nanopart Res 14:1–15. CrossRefGoogle Scholar
  11. Dimkpa CO, Latta DE, McLean JE, Britt DW, Boyanov MI, Anderson AJ (2013) Fate of CuO and ZnO nano-and microparticles in the plant environment. Environ Sci Technol 47:4734–4742. CrossRefPubMedGoogle Scholar
  12. Du W, Gardea-Torresdey JL, Ji R, Yin Y, Zhu J, Peralta-Videa JR, Guo H (2015) Physiological and biochemical changes imposed by CeO2 nanoparticles on wheat: a life cycle field study. Environ Sci Technol 49:11884–11893. CrossRefPubMedGoogle Scholar
  13. Ghanati F, Bakhtiarian S (2014) Effect of methyl jasmonate and silver nanoparticles on production of secondary metabolites by Calendula officinalis L (Asteraceae). Trop J Pharm Res 13:1783–1789. CrossRefGoogle Scholar
  14. Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem 48:909–930. CrossRefGoogle Scholar
  15. Grace SC, Logan BA (2000) Energy dissipation and radical scavenging by the plant phenylpropanoid pathway. Philos Trans R Soc Lond B 355:1499–1510. CrossRefGoogle Scholar
  16. Heo HJ, Kim YJ, Chung D, Kim D-O (2007) Antioxidant capacities of individual and combined phenolics in a model system. Food Chem 104:87–92. CrossRefGoogle Scholar
  17. Hong J, Rico CM, Zhao L, Adeleye AS, Keller AA, Peralta-Videa JR, Gardea-Torresdey JL (2015) Toxic effects of copper-based nanoparticles or compounds to lettuce (Lactuca sativa) and alfalfa (Medicago sativa). Environ Sci Proc Impacts 17:177–185. CrossRefGoogle Scholar
  18. Hou J, Wang X, Hayat T, Wang X (2017) Ecotoxicological effects and mechanism of CuO nanoparticles to individual organisms. Environ Pollut 221:209–217. CrossRefPubMedGoogle Scholar
  19. Iravani S (2011) Green synthesis of metal nanoparticles using plants. Green Chem 13:2638–2650. CrossRefGoogle Scholar
  20. Javed R, Mohamed A, Yücesan B, Gürel E, Kausar R, Zia M (2017) CuO nanoparticles significantly influence in vitro culture, steviol glycosides, and antioxidant activities of Stevia rebaudiana Bertoni. Plant Cell Tiss Organ Cult 131:611–620. CrossRefGoogle Scholar
  21. Khamidov I, Aripova S, Karimov A, Yusupov M (1997) Berberis alkaloids. XL. An investigation of the alkaloids of Berberis thunbergii. Chem Nat Compd 33:599–599. CrossRefGoogle Scholar
  22. Khataee A, Movafeghi A, Mojaver N, Vafaei F, Tarrahi R, Dadpour MR (2017) Toxicity of copper oxide nanoparticles on Spirodela polyrrhiza: assessing physiological parameters. Res Chem Intermed 43:927–941. CrossRefGoogle Scholar
  23. Khodakovskaya MV, de Silva K, Biris AS, Dervishi E, Villagarcia H (2012) Carbon nanotubes induce growth enhancement of tobacco cells. ACS Nano 6:2128–2135. CrossRefPubMedGoogle Scholar
  24. Kim S, Lee S, Lee I (2012) Alteration of phytotoxicity and oxidant stress potential by metal oxide nanoparticles in Cucumis sativus. Water Air Soil Pollut 223:2799–2806. CrossRefGoogle Scholar
  25. Krishnaraj C, Jagan E, Ramachandran R, Abirami S, Mohan N, Kalaichelvan P (2012) Effect of biologically synthesized silver nanoparticles on Bacopa monnieri (Linn.) Wettst. plant growth metabolism. Process Biochem 47:651–658. CrossRefGoogle Scholar
  26. Li A-R, Zhu Y, Li X-N, Tian X-J (2007) Antimicrobial activity of four species of Berberidaceae. Fitoterapia 78:379–381. CrossRefPubMedGoogle Scholar
  27. Mhamdi A, Queval G, Chaouch S, Vanderauwera S, Van Breusegem F, Noctor G (2010) Catalase function in plants: a focus on Arabidopsis mutants as stress-mimic models. J Exp Bot 61:4197–4220. CrossRefGoogle Scholar
  28. Michalak A (2006) Phenolic compounds and their antioxidant activity in plants growing under heavy metal stress. Pol J Environ Stud 15:523–530Google Scholar
  29. Nasrollahzadeh M, Maham M, Sajadi SM (2015) Green synthesis of CuO nanoparticles by aqueous extract of Gundelia tournefortii and evaluation of their catalytic activity for the synthesis of N-monosubstituted ureas and reduction of 4-nitrophenol. J Colloid Interface Sci 455:245–253. CrossRefPubMedGoogle Scholar
  30. Padil VVT, Černík M (2013) Green synthesis of copper oxide nanoparticles using gum karaya as a biotemplate and their antibacterial application. Int J Nanomedicine 8:889–898. CrossRefPubMedCentralGoogle Scholar
  31. Peng C et al (2015) Translocation and biotransformation of CuO nanoparticles in rice (Oryza sativa L.) plants. Environ Pollut 197:99–107. CrossRefPubMedGoogle Scholar
  32. Pietrowska-Borek M, Chadzinikolau T, Kozlowska M (2010) Effect of urban pollution on 4-coumarate: CoA ligase and flavonoid accumulation in Berberis thunbergii. Dendrobiology 64:79–85Google Scholar
  33. Pissuwan D, Niidome T, Cortie MB (2011) The forthcoming applications of gold nanoparticles in drug and gene delivery systems. J Control Release 149:65–71. CrossRefPubMedGoogle Scholar
  34. Poborilova Z, Opatrilova R, Babula P (2013) Toxicity of aluminium oxide nanoparticles demonstrated using a BY-2 plant cell suspension culture model. Environ Exp Bot 91:1–11. CrossRefGoogle Scholar
  35. Prakash MG, Chung IM (2016) Determination of zinc oxide nanoparticles toxicity in root growth in wheat (Triticum aestivum L.) seedlings. Acta Biol Hung 67:286–296. CrossRefPubMedGoogle Scholar
  36. Quettier-Deleu C et al (2000) Phenolic compounds and antioxidant activities of buckwheat (Fagopyrum esculentum Moench) hulls and flour. J Ethnopharmacol 72:35–42. CrossRefPubMedGoogle Scholar
  37. Rai V, Vajpayee P, Singh SN, Mehrotra S (2004) Effect of chromium accumulation on photosynthetic pigments, oxidative stress defense system, nitrate reduction, proline level and eugenol content of Ocimum tenuiflorum L. Plant Sci 167:1159–1169. CrossRefGoogle Scholar
  38. Rico C, Peralta-Videa J, Gardea-Torresdey J (2015) Chemistry, biochemistry of nanoparticles, and their role in antioxidant defense system in plants. In: Siddiqui MH, Al-Whaib M (eds) Nanotechnology and plant sciences. Springer, Cham, pp 1–17Google Scholar
  39. Saif S, Tahir A, Asim T, Chen Y (2016) Plant mediated green synthesis of CuO nanoparticles: comparison of toxicity of engineered and plant mediated CuO nanoparticles towards Daphnia magna. Nanomaterials 6:1–15. CrossRefGoogle Scholar
  40. Sharma P, Jha AB, Dubey RS, Pessarakli M (2012) Reactive oxygen species, oxidative damage, and antioxidative defense mechanism in plants under stressful conditions. J Bot. CrossRefGoogle Scholar
  41. Shaw AK, Hossain Z (2013) Impact of nano-CuO stress on rice (Oryza sativa L.) seedlings. Chemosphere 93:906–915. CrossRefPubMedGoogle Scholar
  42. Shi J et al (2014) Phytotoxicity and accumulation of copper oxide nanoparticles to the Cu-tolerant plant Elsholtzia splendens. Nanotoxicology 8:179–188. CrossRefPubMedGoogle Scholar
  43. Simon L, Fatrai Z, Jonas D, Matkovics B (1974) Study of peroxide metabolism enzymes during the development of Phaseolus vulgaris. Biochem Physiol Pflanz 166:387–392. CrossRefGoogle Scholar
  44. Singleton VL, Orthofer R, Lamuela-Raventós RM (1999) [14] Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. Methods Enzymol 299:152–178. CrossRefGoogle Scholar
  45. Sobkowiak R, Deckert J (2003) Cadmium-induced changes in growth and cell cycle gene expression in suspension-culture cells of soybean. Plant Physiol Biochem 41:767–772. CrossRefGoogle Scholar
  46. Song U, Jun H, Waldman B, Roh J, Kim Y, Yi J, Lee EJ (2013) Functional analyses of nanoparticle toxicity: a comparative study of the effects of TiO2 and Ag on tomatoes (Lycopersicon esculentum). Ecotoxicol Environ Saf 93:60–67. CrossRefPubMedGoogle Scholar
  47. Song G et al (2016) Effects of CuO nanoparticles on Lemna minor. Bot Stud 57:3. CrossRefPubMedPubMedCentralGoogle Scholar
  48. Tarrahi R, Khataee A, Movafeghi A, Rezanejad F, Gohari G (2017) Toxicological implications of selenium nanoparticles with different coatings along with Se 4+ on Lemna minor. Chemosphere 181:655–665. CrossRefPubMedGoogle Scholar
  49. Tran TH, Nguyen VT (2014) Copper oxide nanomaterials prepared by solution methods, some properties, and potential applications: a brief review. Int Sch Res Notices. CrossRefPubMedPubMedCentralGoogle Scholar
  50. Velikova V, Yordanov I, Edreva A (2000) Oxidative stress and some antioxidant systems in acid rain-treated bean plants: protective role of exogenous polyamines. Plant Sci 151:59–66. CrossRefGoogle Scholar
  51. Villinski J, Dumas E, Chai H-B, Pezzuto J, Angerhofer C, Gafner S (2003) Antibacterial activity and alkaloid content of Berberis thunbergii, Berberis vulgaris and Hydrastis canadensis. Pharm Biol 41:551–557. CrossRefGoogle Scholar
  52. Wang Z, Xie X, Zhao J, Liu X, Feng W, White JC, Xing B (2012) Xylem-and phloem-based transport of CuO nanoparticles in maize (Zea mays L.). Environ Sci Technol 46:4434–4441. CrossRefPubMedGoogle Scholar
  53. Wang H et al (2013) Graphene-based materials: fabrication, characterization and application for the decontamination of wastewater and wastegas and hydrogen storage/generation. Adv Colloid Interface Sci 195:19–40. CrossRefPubMedGoogle Scholar
  54. Winterbourn CC, McGrath BM, Carrell RW (1976) Reactions involving superoxide and normal and unstable haemoglobins. Biochem J 155:493–502. CrossRefPubMedPubMedCentralGoogle Scholar
  55. Zafar H, Ali A, Ali JS, Haq IU, Zia M (2016) Effect of ZnO nanoparticles on Brassica nigra seedlings and stem explants: growth dynamics and antioxidative response. Front Plant Sci 7:535. CrossRefPubMedPubMedCentralGoogle Scholar
  56. Zhang Q et al (2014) CuO nanostructures: synthesis, characterization, growth mechanisms, fundamental properties, and applications. Prog Mater Sci 60:208–337. CrossRefGoogle Scholar
  57. Zhao J et al (2017) Uptake, distribution and transformation of CuO NPs in a floating plant Eichhornia crassipes and related stomatal responses. Environ Sci Technol 51:7686–7695. CrossRefPubMedGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Department of Plant Biology, Faculty of Natural SciencesUniversity of TabrizTabrizIran
  2. 2.Department of Inorganic Chemistry, Faculty of ChemistryTabriz UniversityTabrizIran
  3. 3.Drug Applied Research CenterTabriz University of Medical SciencesTabrizIran

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