Biochemical Responses of Duckweed (Spirodela polyrhiza) to Zinc Oxide Nanoparticles

  • Changwei Hu
  • Yimeng Liu
  • Xiuling Li
  • Mei LiEmail author


The present study focuses on the biochemical responses of the aquatic plant duckweed (Spirodela polyrhiza L.) to zinc oxide nanoparticles (ZnO NPs). Laboratory experiments were performed using a 96-h exposure to 25-nm NPs at different concentrations (0, 1, 10, and 50 mg/L). Growth, chlorophyll-to-pheophytin ratio (D665/D665a) and activities of superoxide dismutase, catalase, peroxidase (POD), and Na+, K+-ATPase were determined as indices to evaluate the toxicity of NPs in the culture medium. To understand better whether the Zn2+ released from the ZnO NP suspensions plays a key role in toxicity of the NPs, we investigated particle aggregation and dissolution in the medium. Furthermore, two exposure treatments for the group with the highest concentration (50 mg/L) were performed: (1) exposure for the full 96 h (50a treatment) and (2) the medium being replaced with culture medium without NPs after 12 h (50b treatment). Our results indicate that ZnO NPs induced adverse effects in S. polyrhiza at the concentration of 50 mg/L in the culture medium. Zn2+ released from the NPs might be the main source of its toxicity to this species.


Frond Number Spirodela Polyrhiza Duckweed Growth Consumer Product Inventory Physiological Stress Index 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



This work was supported by Shandong Outstanding Young Scientist Award Fund (Grant No. BS2010SF005), Dr. Start Fund of Linyi University (Grant No. BS201009), and a Project of Shandong Province Higher Educational Science and Technology Program (Grant No. J10LE57). The authors are grateful to Qingcai Xu for ion determination and Richard A. Manderville for assistance and helpful advice on the manuscript.


  1. Adams LK, Lyon DY, Alvarez PJJ (2006) Comparative ecotoxicity of nanoscale TiO2, SiO2 and ZnO water suspensions. Water Res 40:3527–3532CrossRefGoogle Scholar
  2. Aruoja V, Dubourguier HC, Kasemets K, Kahru A (2009) Toxicity of nanoparticles of CuO, ZnO and TiO2 to microalgae Pseudokirchneriella subcapitata. Sci Total Environ 407:1461–1468CrossRefGoogle Scholar
  3. Balen B, Tkalec M, Sikic S, Tolic S, Cvjetko P, Pavlica M et al (2011) Biochemical responses of Lemna minor experimentally exposed to cadmium and zinc. Ecotoxicology 20:815–826CrossRefGoogle Scholar
  4. Borm P, Klaessig FC, Landry TD, Moudgil B, Pauluhn J, Thomas K et al (2006) Research strategies for safety evaluation of nanomaterials. Part V: role of dissolution in biological fate and effects of nanoscale particles. Toxicol Sci 90:23–32CrossRefGoogle Scholar
  5. Boxall ABA, Chaudhry Q, Sinclair C, Jones A, Aitken R, Jefferson B, et al. C (2007) Current and future predicted environmental exposure to engineered nanoparticles. Central Science Laboratory, Department of the Environment and Rural Affairs, London, UKGoogle Scholar
  6. Brayner R (2008) The toxicological impact of nanoparticles. Nanotoday 3:48–55Google Scholar
  7. Brunauer S, Emmett PH, Teller E (1938) Adsorption of gases in multimolecular layers. J Am Chem Soc 60:309–319CrossRefGoogle Scholar
  8. Buffle J, Leppard GG (1995) Characterization of aquatic colloids and macromolecules. 1. Structure and behavior of colloidal material. Environ Sci Technol 29:2169–2175CrossRefGoogle Scholar
  9. Cakmak I (2000) Possible roles of zinc in protecting plant cells from damage by reactive oxygen species. New Phytol 146:185–205CrossRefGoogle Scholar
  10. Chang JS, Chang KLB, Hwang DF, Kong ZL (2007) In vitro cytotoxicity of silica nanoparticles at high concentrations strongly depends on the metabolic activity type of the cell line. Environ Sci Technol 41:2064–2068CrossRefGoogle Scholar
  11. Charpentier J, Garnier J (1985) Etude de la multiplication et de la formation des colonies de Spirodela polyrhiza [in French]. Comptes Rendus de l'Académie des Sciences 300:587–590Google Scholar
  12. Charpentier J, Garnier J, Flaugniatti R (1987) Toxicology and bioaccumulation of cadmium in experimental cultures of duckweed Lemna polyrhiza. Bull Environ Contam Toxicol 38:1055–1061CrossRefGoogle Scholar
  13. Dang Z, Lock RA, Flik G, Wendelaar Bonga SE (2000) Na+/K+-ATPase immunoreactivity in branchial chloride of Oreochromis mossambicus exposed to copper. J Exp Biol 203:379–387Google Scholar
  14. Di Toro DM, Zarba CS, Hansen DJ, Berry WJ, Swartz RC (1991) Technical basis for establishing sediment quality criteria for nonionic organic chemicals using equilibrium partitioning. Environ Toxicol Chem 10:1541–1583CrossRefGoogle Scholar
  15. Farré M, Gajda-Schrantz K, Kantiani L, Barceló D (2009) Ecotoxicity and analysis of nanomaterials in the aquatic environment. Anal Bioanal Chem 393:81–95CrossRefGoogle Scholar
  16. Franklin NM, Rogers NJ, Apte SC, Batley GE, Gadd GE, Casey PS (2007) Comparative toxicity of nanoparticulate ZnO, Bulk ZnO, and ZnCl2 to a freshwater microalga (Pseudokirchneriella subcapitata): the importance of particle solubility. Environ Sci Technol 41:8484–8490CrossRefGoogle Scholar
  17. Gubbins EJ, Batty LC, Lead JR (2011) Phytotoxicity of silver nanoparticles to Lemna minor L. Environ Pollut 159:1551–1559CrossRefGoogle Scholar
  18. Gustafsson O, Gschwemd G (1997) Aquatic colloids: concepts, definitions, and current challenges. Limnol Oceanogr 42:519–528CrossRefGoogle Scholar
  19. Heinlaan M, Ivask A, Blinova I, Dubourguier HC, Kahru A (2008) Toxicity of nanosized and bulk ZnO, CuO and TiO2 to bacteria Vibrio fischeri and crustaceans Daphnia magna and Thamnocephalus platyurus. Chemosphere 71:1308–1316CrossRefGoogle Scholar
  20. Hoagland DR, Arnon DI (1938) The water culture method for growing plants without soil. California Agriculture Experimental Station Circular 347, Berkeley, CAGoogle Scholar
  21. Hu X, Cook S, Wang P, Hwang H (2009) In vitro evaluation of cytotoxicity of engineered metal oxide nanoparticles. Sci Total Environ 407:3070–3072CrossRefGoogle Scholar
  22. Huff EA (2011) Untested nanoparticles showing up in thousands of consumer products. Available at: Accessed:
  23. Kahru A, Dubourguier HC (2010) From ecotoxicology to nanoecotoxicology. Toxicology 269:105–119CrossRefGoogle Scholar
  24. Lecoanet HF, Wiesner MR (2004) Velocity effects on fullerene and oxide nanoparticle deposition in porous media. Environ Sci Technol 38:4377–4382CrossRefGoogle Scholar
  25. Lecoanet HF, Bottero JY, Wiesner MR (2004) Laboratory assessment of the mobility of nanomaterials in porous media. Environ Sci Technol 38:5164–5169CrossRefGoogle Scholar
  26. Li HS (2000) Principles and techniques of plant physiological biochemical experiment [in Chinese]. Beijing Higher Education Press, Beijing, pp 134–137Google Scholar
  27. Lin DH, Xing BS (2008) Root uptake and phytotoxicity of ZnO nanoparticles. Environ Sci Technol 42:5580–5585CrossRefGoogle Scholar
  28. Lopez J, Retuerto R, Carballeira A (1997) D665/D665a index vs. frequencies as indicators of bryophyte response to physicochemical gradients. Ecology 78:261–271Google Scholar
  29. Masciangioli T, Zhang WX (2003) Environmental technologies at the nanoscale. Environ Sci Technol 1:102–108CrossRefGoogle Scholar
  30. Maynard A, Michelson E (2006) The Nanotechnology Consumer Products Inventory. Woodrow Wilson International Center for Scholars. Available at: Accessed:
  31. McCord JM, Fridovich I (1969) Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein). J Biol Chem 244:6049–6055Google Scholar
  32. Megateli S, Semsari S, Couderchet M (2009) Toxicity and removal of heavy metals (cadmium, copper, and zinc) by Lemna gibba. Ecotoxicol Environ Safe 72:1774–1780CrossRefGoogle Scholar
  33. Nel A, Xia T, Madler L, Li N (2006) Toxic potential of materials at the nanolevel. Science 311:622–627CrossRefGoogle Scholar
  34. Peralta-Videa JR, Zhao LJ, Lopez-Moreno ML, de la Rosa G, Hong J, Gardea-Torresdey JL (2011) Nanomaterials and the environment: a review for the biennium 2008–2010. J Hazard Mater 186:1–15CrossRefGoogle Scholar
  35. Rao MV, Paliyath G, Ormrod DP (1996) Ultraviolet-B and ozone-induced biochemical changes in antioxidant enzymes of Arabidopsis thaliana. Plant Physiol 110:125–136CrossRefGoogle Scholar
  36. Shi JY, Abid AD, Kennedy IM, Hristova KR, Silk WK (2011) To duckweeds (Landoltia punctata), nanoparticulate copper oxide is more inhibitory than the soluble copper in the bulk solution. Environ Pollut 159:1277–1282CrossRefGoogle Scholar
  37. Shiosaka T, Okuda H, Fungi S (1971) Mechanisms of phosphorylation of thymidine by the culture filtrate of Clostridium perfringens and rat liver extract. Biochim Biophys Acta 246:171–183CrossRefGoogle Scholar
  38. Song Y, Zhu LS, Wang J, Wang JH, Liu W, Xie H (2008) DNA damage and effects on antioxidative enzymes in earthworm (Eisenia fetida) induced by atrazine. Soil Biol Biochem 41:905–909CrossRefGoogle Scholar
  39. Sun O, Wang Q, Jena P, Kawazoe Y (2005) Clustering of Ti on a C-60 surface and its effect on hydrogen storage. J Am Chem Soc 127:14582–14583CrossRefGoogle Scholar
  40. United States Environmental Protection Agency (2007) Final nanotechnology white paper. EPA 100/B-07/001, USEPA, Washington, DCGoogle Scholar
  41. Upadhyay R, Panda SK (2010) Zinc reduces copper toxicity induced oxidative stress by promoting antioxidant defense in freshly grown aquatic duckweed Spirodela polyrhiza L. J Hazard Mater 175:1081–1084CrossRefGoogle Scholar
  42. Weckx J, Clijsters H (1996) Oxidative damage and defense mechanisms in primary leaves of Phaseolus vulgaris as a result of root assimilation of toxic amounts of copper. Physiol Plantarum 96:506–512CrossRefGoogle Scholar
  43. Wu B, Wang Y, Lee YH, Horst A, Wang ZP, Chen DR et al (2010) Comparative eco-toxicities of nano-ZnO particles under aquatic and aerosol exposure modes. Environ Sci Technol 44:1484–1489CrossRefGoogle Scholar
  44. Zhu MT, Feng WY, Wang B, Wang TC, Gu YQ, Wang M et al (2008a) Comparative study of pulmonary responses to nano- and submicron-sized ferric oxide in rats. Toxicology 247:102–111CrossRefGoogle Scholar
  45. Zhu YG, Sun GX, Lei M, Teng M, Liu YX, Chen NC et al (2008b) High percentage inorganic arsenic content of mining impacted and nonimpacted Chinese rice. Environ Sci Technol 42:5008–5013CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2012

Authors and Affiliations

  1. 1.Shandong Provincial Key Laboratory of Water and Soil Conservation and Environmental ProtectionLinyi UniversityLinyiChina
  2. 2.School of Environmental Science and EngineeringShandong UniversityJinanChina
  3. 3.State Key Laboratory of Pollution Control and Resource ReuseSchool of the Environment, Nanjing UniversityNanjingChina

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