, Volume 23, Issue 7, pp 1292–1304 | Cite as

Pb-inhibited mitotic activity in onion roots involves DNA damage and disruption of oxidative metabolism

  • Gurpreet Kaur
  • Harminder Pal Singh
  • Daizy Rani Batish
  • Ravinder Kumar Kohli


Plant responses to abiotic stress significantly affect the development of cells, tissues and organs. However, no studies correlating Pb-induced mitotic inhibition and DNA damage and the alterations in redox homeostasis during root division per se were found in the literature. Therefore, an experiment was conducted to evaluate the impact of Pb on mitotic activity and the associated changes in the oxidative metabolism in onion roots. The cytotoxic effect of Pb on cell division was assessed in the root meristems of Allium cepa (onion). The mitotic index (MI) was calculated and chromosomal abnormalities were sought. Pb-treatment induced a dose-dependent decrease in MI in the onion root tips and caused mitotic abnormalities such as distorted metaphase, fragments, sticky chromosomes, laggards, vagrant chromosomes and bridges. Single Cell Gel Electrophoresis was also performed to evaluate Pb induced genotoxicity. It was accompanied by altered oxidative metabolism in the onion root tips suggesting the interference of Pb with the redox homeostasis during cell division. There was a higher accumulation of malondialdehyde, conjugated dienes and hydrogen peroxide, and a significant increase in the activities of superoxide dismutases, ascorbate peroxidases, guaiacol peroxidases and glutathione reductases in Pb-treated onion roots, whereas catalases activity exhibited a decreasing pattern upon Pb exposure. The study concludes that Pb-induced cytotoxicity and genotoxicity in the onion roots is mediated through ROS and is also tightly linked to the cell cycle. The exposure to higher concentrations arrested cell cycle leading to cell death, whereas different repair responses are generated at lower concentrations, thereby allowing the cell to complete the cell cycle.


Pb toxicity Chromosomal abnormalities Mitotic index (MI) Genotoxicity DNA damage Redox homeostasis 



Gurpreet Kaur is thankful to University Grants Commission, New Delhi, India, for financial assistance in the form of Post-doctoral Fellowship.

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Achary VMM, Panda BB (2010) Aluminium-induced DNA damage and adaptive response to genotoxic stress in plant cells are mediated through reactive oxygen intermediates. Mutagenesis 25:201–209CrossRefGoogle Scholar
  2. Achary VMM, Jena S, Panda KK, Panda BB (2008) Aluminium induced oxidative stress and DNA damage in root cells of Allium cepa L. Ecotoxicol Environ Safe 70:300–310CrossRefGoogle Scholar
  3. Awashthi SK (2000) Prevention of Food Adulteration Act no 37 of 1954. Central and State Rules as Amended for 1999. Ashoka Law House, New Delhi, IndiaGoogle Scholar
  4. Băra CI, Capraru G, Băra II, Cîmpeanu MM, Buburuzan L, Axente FM (2004) The effects of treatment with lead acetate on mitosis at Larix decidua L and Picea abies L. Ann Univ Alexandru Ioan Cuza Iaşi (N S). Genet Mol Biol Tom V: 180–187Google Scholar
  5. Chaney RL, Sterrett SB, Mielke HW (1984) The potential for heavy metal exposure from urban gardens and soils. In: Preer JR (ed) Proceedings of the symposium on heavy metals in urban gardens, Agricultural Experiment Station. University of the District of Columbia, Washington, pp 37–84Google Scholar
  6. Chang XM, Wang WC (1991) DNA amplifications, chromatin variations and polytene chromosomes in differentiating cell of common bread wheat in vitro and root of regenerated plants in vivo. Genome 34:799–809CrossRefGoogle Scholar
  7. Darlington CD, Mc Leish L (1951) Action of maleic hydrazide on the cell. Nature 167:407–408CrossRefGoogle Scholar
  8. Duez P, Dehon G, Kumps A, Dubois J (2003) Statistics of the Comet assay: a key to discriminate between genotoxic effects. Mutagenesis 18:159–166CrossRefGoogle Scholar
  9. Erturk FA, Ay H, Nardemir G, Agar G (2013) Molecular determination of genotoxic effects of cobalt and nickel on maize (Zea mays L.) by RAPD and protein analyses. Toxicol Ind Health 29:662–671CrossRefGoogle Scholar
  10. Eun SO, Youn HS, Lee Y (2000) Lead disturbs microtubule organization in the root meristem of Zea mays. Physiol Plant 110:357–365CrossRefGoogle Scholar
  11. European Union (2002) Heavy metals in wastes. European commission on environment
  12. Ezaki B, Suzuki M, Motoda H, Kawamura M, Nakashima S, Matsumoto H (2004) Mechanism of gene expression of Arabidopsis Glutathione S-Transferase, AtGST1, and AtGST11 in response to aluminum stress. Plant Physiol 134:1672–1682CrossRefGoogle Scholar
  13. Fiskesjö G (1985) The Allium test as a standard in environmental monitoring. Hereditas 102:99–112CrossRefGoogle Scholar
  14. Fiskesjö G (1997) Allium test for screening chemicals: evaluation of cytologic parameters. In: Wang W, Gorsuch JW, Hughes JS (eds) Plants for environmental studies. CRC Publishers, Boca Raton, pp 308–333Google Scholar
  15. Fusconi A, Repetto O, Bona E, Massa N, Gallo C, Dumas-Gaudot E, Berta G (2006) Effects of Cadmium on meristem activity and nucleus ploidy in roots of Pisum sativum L. cv. Frisson seedlings. Environ Exp Bot 58:253–260CrossRefGoogle Scholar
  16. Gajewska E, Słaba M, Andrzejewska R, Skłodowska M (2006) Nickel-induced inhibition of wheat root growth is related to H2O2 production but not to lipid peroxidation. Plant Growth Regul 49:95–103Google Scholar
  17. Gichner T, Patková Z, Száková J, Demnerová K (2006) Toxicity and DNA damage in tobacco and potato plants growing on soil polluted with heavy metals. Ecotoxicol Environ Safe 65(3):420–426CrossRefGoogle Scholar
  18. Gupta DK, Nicoloso FT, Schetinger MRC, Rossato LV, Pereira LB, Castro GY, Srivastava S, Tripathi RD (2009) Antioxidant defense mechanism in hydroponically grown Zea mays seedlings under moderate lead stress. J Hazard Mater 172:479–484CrossRefGoogle Scholar
  19. Hartwig A (1994) Role of DNA repair inhibition in Lead- and Cadmium-induced genotoxicity: a review. Environ Health Persp 102:45–50CrossRefGoogle Scholar
  20. Hepler PK (1992) Calcium and mitosis. Int Rev Cytol 138:239–267CrossRefGoogle Scholar
  21. Holmgren G, Meyer M, Chaney R, Daniels R (1993) Cadmium, lead, zinc, copper, and nickel in agricultural soils of the United States of America. J Environ Qual 22:335–348CrossRefGoogle Scholar
  22. Kasai H, Yamaizumi Z, Berger M, Cadet J (1992) Photosensitized formation of 7,8-dihydro-8-oxo-2′-deoxyguanosine (8-hydroxy-2′-deoxyguanosine) in DNA by riboflavin: a non singlet oxygen-mediated reaction. J Am Chem Soc 114:9642–9644Google Scholar
  23. Kaur G, Singh HP, Batish DR, Kohli RK (2012) Growth, photosynthetic activity and oxidative stress in wheat (Triticum aestivum) after exposure of lead to soil. J Environ Biol 33:265–269Google Scholar
  24. Kaur G, Singh HP, Batish DR, Kohli RK (2013) Lead (Pb)-induced biochemical and ultrastructural changes in wheat (Triticum aestivum) roots. Protoplasma 250:53–62CrossRefGoogle Scholar
  25. Końca K, Lankoff A, Banasik A, Lisowska H, Kuszewski T, Góźdź S, Koza Z, Wojcik A (2003) A cross platform public domain PC image analysis program for the comet assay. Mutat Res 534:15–20CrossRefGoogle Scholar
  26. Kozhevnikova AD, Seregin IV, Bystrova EI, Belyaeva AI, Kataeva MN, Ivanov VB (2009) The effects of Lead, Nickel, and Strontium nitrates on cell division and elongation in maize roots. Russ J Plant Physiol 56:242–250Google Scholar
  27. Kumari M, Mukherjee A, Chandrasekaran N (2009) Genotoxicity of silver nanoparticles in Allium cepa. Sci Total Environ 407:5243–5246CrossRefGoogle Scholar
  28. Kuras M, Pilarski R, Nowakowska J, Zobel A, Brzost K, Antosiewicz J, Gulewicz K (2009) Effect of alkaloid free and alkaloid rich preparations from Uncaria tomentosa bark on mitotic activity and chromosome morphology evaluated by Allium test. J Ethnopharmacol 121:140–147CrossRefGoogle Scholar
  29. Li H, Wang Q, Cui Y, Dong Y, Christie P (2005) Slow release chelate enhancement of lead phytoextraction by corn (Zea mays L.) from contaminated soil a preliminary study. Sci Total Environ 339:179–187CrossRefGoogle Scholar
  30. Liu D, Zou J, Meng Q, Zou J, Jiang W (2009) Uptake and accumulation and oxidative stress in garlic (Allium sativum L.) under lead phytotoxicity. Ecotoxicology 18:134–143CrossRefGoogle Scholar
  31. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein estimation with Folin-phenol reagent. J Biol Chem 193:265–278Google Scholar
  32. Ma TH (1982) Tradescantia cytogenetic tests (root tip mitosis, pollen mitosis, pollen mother meiosis). A report of the US Environmental Protection Agency Gene-Tox program. Mutat Res 99:293–302CrossRefGoogle Scholar
  33. Malecka A, Piechalak A, Tomaszewska B (2009) Reactive oxygen species production and antioxidative defense system in pea root tissues treated with lead ions: the whole roots level. Acta Physiol Plant 31:1053–1063Google Scholar
  34. Małecka A, Piechalak A, Morkunas I, Tomaszewska B (2008) Accumulation of lead in root cells of Pisum sativum. Acta Physiol Plant 30:629–637Google Scholar
  35. May MJ, Vernoux T, Leaver C, Van Montagu M, Inzé D (1998) Glutathione homeostasis in plants: implications for environmental sensing and plant development. J Exp Bot 49:649–667Google Scholar
  36. Meyers DER, Auchterlonie GJ, Webb RI, Wood B (2008) Uptake and localisation of lead in the root system of Brassica juncea. Environ Pollut 153:323–332CrossRefGoogle Scholar
  37. Nagajyoti PC, Lee KD, Sreekanth TVM (2010) Heavy metals, occurrence and toxicity for plants: a review. Environ Chem Lett 8:199–216CrossRefGoogle Scholar
  38. Nanushyan ER, Murashev VV (2003) Induction of multinuclear cells in the apical meristems of Allium cepa by geomagnetic field outrages. Russ J Plant Physiol 50:522–526Google Scholar
  39. National Research Council. 1980. Lead in the human environment. Report Number PB-82-117136. National Academy of Sciences, Washington DCGoogle Scholar
  40. Pădureanu S (2005) Cytogenetics effects induced by nitrate of lead on mitotic division at Allium cepa L. The Scientific Annals of “Al. I. Cuza” University, Genet Biol Mol Tom VI: 201–206Google Scholar
  41. Pakrashi S, Jain N, Dalai S, Jayakumar J, Chandrasekaran PT, Raichir AM, Chandrasekaran N, Mukherjee A (2014) In vivo genotoxicity assessment of titanium dioxide nanoparticles by Allium cepa root tip assay at high exposure concentrations. PLoS One 9(2):e87789CrossRefGoogle Scholar
  42. Panda BB, Panda KK (2002) Genotoxicity and mutagenicity of metals in plants. In: Prasad MNV, Strzalka K (eds) Physiology and biochemistry of metal tolerance in plants. Kluwer Academic Publishers, Amsterdam, pp 395–414CrossRefGoogle Scholar
  43. Phugare SS, Kalyani DC, Patil AV, Jadhav JP (2011) Textile dye degradation by bacterial consortium and subsequent toxicological analysis of dye and dye metabolites using cytotoxicity, genotoxicity and oxidative stress studies. J Hazard Mater 186:713–723CrossRefGoogle Scholar
  44. Piechalak A, Tomaszewska B, Baralkiewicz D, Malecka A (2002) Accumulation and detoxification of lead ions in legumes. Phytochemistry 60:153–162CrossRefGoogle Scholar
  45. Poli P, Buschini A, Restivo FM, Ficarelli A, Cassoni F, Ferrero I (1999) Comet assay application in environmental monitoring: DNA damage in human leukocytes and plant cells in comparison with bacterial and yeast tests. Mutagenesis 14:547–555CrossRefGoogle Scholar
  46. Pourrut B, Jean S, Silvestre J, Pinelli E (2011) Lead-induced DNA damage in Vicia faba root cells: potential involvement of oxidative stress. Mutat Res 726:123–128CrossRefGoogle Scholar
  47. Qureshi MI, Abdin MZ, Qadir S, Iqbal M (2007) Lead induced oxidative stress and metabolic alterations in Cassia angustifolia Vahl. Biol Plant 51:121–128CrossRefGoogle Scholar
  48. Radetski CM, Ferrari B, Cotelle S, Masfaraud JF, Ferard JF (2004) Evaluation of genotoxic, mutagenic and oxidative stress potentials of municipal soild waste incinerator bottom ash leachates. Sci Total Environ 333:204–221CrossRefGoogle Scholar
  49. Reddy AM, Kumar SG, Jyothsnakumari G, Thimmanaik S, Sudhakar C (2005) Lead induced changes in antioxidant metabolism of horsegram (Macrotyloma uniflorum (Lam.) Verdc.) and bengalgram (Cicer arietinum L.). Chemosphere 60:97–104CrossRefGoogle Scholar
  50. Reichheld JP, Vernoux T, Lardon F, Van Montagu M, Inzé D (1999) Specific checkpoints regulate plant cell cycle progression in response to oxidative stress. Plant J 17:647–656CrossRefGoogle Scholar
  51. Ryan J, Scheckel K, Berti W, Brown S, Casteel S, Cheney R, Hallfrisch J, Doolan M, Grevati P, Maddaloni M, Mosby D (2004) Reducing children’s risk from lead in soil. Environ Sci Technol 38:19A–24ACrossRefGoogle Scholar
  52. Santoro A, Lioi MB, Monfregola J, Salzano S, Barbieri R, Ursisni MV (2005) L-Carnitine protects mammalian cells from chromosome aberrations but not from inhibition of cell proliferation induced by hydrogen peroxide. Mutat Res 587:16–25CrossRefGoogle Scholar
  53. Seregin IV, Ivanov VB (2001) Physiological aspects of cadmium and lead toxic effects on higher plants. Russ J Plant Physiol 48:523–544Google Scholar
  54. Siddiqui S (2012) Lead induced genotoxicity in Vigna mungo var: hD-94. J Saudi Soc Agric Sci 11:107–112Google Scholar
  55. Singh HP, Batish DR, Kaur S, Setia N, Kohli RK (2005) Effects of 2-benzoxazolinone on the germination, early growth and morphogenetic response of mung bean (Phaseolus aureus). Ann App Biol 147:267–274CrossRefGoogle Scholar
  56. Singh HP, Kaur S, Batish DR, Sharma VP, Sharma N, Kohli RK (2009) Nitric oxide alleviates arsenic toxicity by reducing oxidative damage in the roots of Oryza sativa (rice). Nitric Oxide 20:289–297CrossRefGoogle Scholar
  57. Singh HP, Kaur G, Batish DR, Kohli RK (2011) Lead (Pb)-inhibited radicle emergence in Brassica campestris involves alterations in starch-metabolizing enzymes. Biol Trace Elem Res 144:1295–1301CrossRefGoogle Scholar
  58. Steinkellner H, Mun-Sik K, Helma C, Ecker S, Ma TH, Kundi M, Knasmüller S (1998) Genotoxic effects of heavy metals: comparative investigation with plant bioassays. Environ Mol Mutagen 31:183–191CrossRefGoogle Scholar
  59. Sudhakar R, Gowda N, Venu G (2001) Mitotic abnormalities induced by silk dyeing industry effluents in the cells of Allium cepa. Cytologia 66:235–239CrossRefGoogle Scholar
  60. Tice RR, Agurell E, Anderson D, Burlinson B, Hartmann A, Kobayashi H, Miyamae Y, Rojas E, Ryu JC, Sasaki YF (2000) Single cell gel/Comet Assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ Mol Mutagen 35:206–221CrossRefGoogle Scholar
  61. Ünyayar S, Çelik A, Çekiç FÖ, Gozel A (2006) Cadmium-induced genotoxicity, cytotoxicity and lipid peroxidation in Allium sativum and Vicia faba. Mutagenesis 21:77–81CrossRefGoogle Scholar
  62. Verma S, Dubey RS (2003) Lead toxicity induces lipid peroxidation and alters the activities of antioxidant enzymes in growing rice plants. Plant Sci 164:645–655CrossRefGoogle Scholar
  63. Vidakovic-Cifrek Z, Pavlica M, Regula I, Papes D (2002) Cytogenetic damage in shallot (Allium cepa) root meristems induced by oil industry “High density Brines”. Arch Environ Contam Toxicol 43:284–291CrossRefGoogle Scholar
  64. Werner DA, Edvards GE (1993) Effects of polyploidy on photosynthesis. Photosynth Res 35:135–147CrossRefGoogle Scholar
  65. Wierzbicka M (1999) The effect of lead on the cell cycle in the root meristem of Allium cepa L. Protoplasma 207:186–194CrossRefGoogle Scholar
  66. Yücel E, Hatİpoğlu A, Sözen E, Guner ST (2008) The effects of the lead (PbCl2) on mitotic cell division of Anatolian Black Pine (Pinus nigra ssp. pallasiana). BioDiCon 1(2):124–129Google Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Gurpreet Kaur
    • 1
  • Harminder Pal Singh
    • 1
  • Daizy Rani Batish
    • 2
  • Ravinder Kumar Kohli
    • 2
  1. 1.Department of Environment StudiesPanjab UniversityChandigarhIndia
  2. 2.Department of BotanyPanjab UniversityChandigarhIndia

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