Biochemical Adaptations in Zea mays Roots to Short-Term Pb2+ Exposure: ROS Generation and Metabolism

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


The present study investigated the effect of lead (0, 16, 40 and 80 mg L−1 Pb2+) exposure for 3, 12 and 24 h on root biochemistry in hydroponically grown Zea mays (maize). Pb2+ exposure (80 mg L−1) enhanced malondialdehyde content (239 %–427 %), reactive carbonyl groups (425 %–512 %) and H2O2 (129 %–294 %) accumulation during 3–24 h of treatment, thereby indicating cellular peroxidation and oxidative damage. The quantitative estimations were in accordance with in situ detection of ROS generation (using 2′,7′-dichlorodihydrofluorescein diacetate dye) and H2O2 accumulation. Pb2+ treatment significantly reduced ascorbate and glutathione content during 3–24 h of exposure. On the contrary, levels of non-protein thiols were enhanced by 3–11.8 time over control in response to 16–80 mg L−1 Pb2+ treatment, after 24 h. A dose-dependent induction in ascorbate peroxidase and lipoxygenase enzyme activity was observed in Z. mays roots. The activities of ascorbate-recycling enzymes (dehydroascorbate reductase and monodehydroascorbate reductase) were significantly increased in relation to concentration and duration of Pb2+ treatment. The study concludes that Pb2+-exposure induces ROS-mediated oxidative damage during early period of exposure despite the upregulation of enzymes of ascorbate–glutathione cycle.


Cellular peroxidation ROS accumulation Cell death Defense mechanisms 



GK is thankful to University Grants Commission (New Delhi, India) for research fellowship.

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–399CrossRefGoogle Scholar
  2. Baker CJ, Mock NM (1994) An improved method for monitoring cell death in cell suspension and leaf disc assays using Evans blue. Plant Cell Tissue Organ Cult 39:7–12CrossRefGoogle Scholar
  3. Biteur N, Aoues A, Kharoubi O, Slimani M (2011) Oxidative stress induction by lead in leaves of radish (Raphanus sativus) seedlings. Not Sci Biol 3(4):93–99Google Scholar
  4. Blokhina OB, Virolainen E, Fagerstedt KV (2003) Antioxidants, oxidative damage and oxygen deprivation stress: a review. Ann Bot 91:179–194CrossRefGoogle Scholar
  5. Boscolo PRS, Menossi M, Jorge RA (2003) Aluminum-induced oxidative stress in maize. Phytochemistry 62:181–189CrossRefGoogle Scholar
  6. Chowhan N, Singh HP, Batish DR, Kaur S, Ahuja N, Kohli RK (2013) β-Pinene inhibited germination and early growth involves membrane peroxidation. Protoplasma 250:691–700CrossRefGoogle Scholar
  7. Ellman GL (1959) Tissue sulfhydryl groups. Arch Biochem Biophys 82:70–77CrossRefGoogle Scholar
  8. Ezaki B, Gardner RC, Ezaki Y, Matsumoto H (2000) Expression of aluminum-induced genes in transgenic Arabidopsis plants can ameliorate aluminum stress and/or oxidative stress. Plant Physiol 122:657–665CrossRefGoogle Scholar
  9. Foyer CH, Halliwell B (1976) The presence of glutathione and glutathione reductase in chloroplasts: a proposed role in ascorbic acid metabolism. Planta 133:21–25CrossRefGoogle Scholar
  10. Geebelen W, Vangronsveld J, Adriano DC, Van Poucke LC, Clijsters H (2002) Effects of Pb-EDTA and EDTA on oxidative stress reactions and mineral uptake in Phaseolus vulgaris. Physiol Plant 115(3):377–384CrossRefGoogle Scholar
  11. Griffith OW (1980) Determination of glutathione and glutathione disulfide using glutathione reductase and 2-vinylpyridine. Anal Biochem 106(1):207–212CrossRefGoogle Scholar
  12. 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
  13. Heath RL, Packer L (1968) Photoperoxidation in isolated chloroplasts. I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys 125:189–198CrossRefGoogle Scholar
  14. Hossain MA, Asada K (1984) Purification of dehydroascorbate reductase from spinach and its characterization as a thiol enzyme. Plant Cell Physiol 25:85–92Google Scholar
  15. Hossain MA, Nakano Y, Asada K (1984) Monodehydroascorbate reductase in spinach chloroplasts and its participation in regeneration of ascorbate for scavenging hydrogen peroxide. Plant Cell Physiol 25:385–395Google Scholar
  16. Huang H, Gupta DK, Tian S, Yang X, Li T (2012) Lead tolerance and physiological adaptation mechanism in roots of accumulating and non-accumulating ecotypes of Sedum alfredii. Environ Sci Pollut Res 19:1640–1651CrossRefGoogle Scholar
  17. Kaur G, Singh HP, Batish DR, Kohli RK (2012) A time-course assessment of changes in reactive oxygen species generation and antioxidant defense in hydroponically grown wheat in response to lead ions (Pb2+). Protoplasma 249:1091–1100CrossRefGoogle Scholar
  18. Kaur G, Singh HP, Batish DR, Kohli RK (2014) Pb-inhibited mitotic activity in onion roots involves DNA damage and disruption of oxidative metabolism. Ecotoxicology 23:1292–1304CrossRefGoogle Scholar
  19. Komárek M, Tlustos P, Száková J, Chrastný V, Ettler V (2007) The use of maize and poplar in chelant-enhanced phytoextraction of lead from contaminated agricultural soils. Chemosphere 67:640–651CrossRefGoogle Scholar
  20. Krzesłowska M, Lenartowska M, Samardakiewicz S, Bilski H, Wόzny A (2010) Lead deposited in the cell wall of Funaria hygrometrica protonemata is not stable—a remobilization can occur. Environ Pollut 158(1):325–338CrossRefGoogle Scholar
  21. Kumar A, Prasad MNV, Achary VMM, Panda BB (2013) Elucidation of lead-induced oxidative stress in Talinum triangulare roots by analysis of antioxidant responses and DNA damage at cellular level. Environ Sci Pollut Res 20:4551–4561CrossRefGoogle Scholar
  22. Law MY, Charles SA, Halliwell B (1983) Glutathione and ascorbic acid in spinach (Spinacia oleracea) chloroplasts. The effect of hydrogen peroxide and of Paraquat. Biochem J 210(3):899–903Google Scholar
  23. Levine RL, Williams JA, Stadtman ER, Shacter E (1994) Carbonyl assay for determination of oxidatively modified proteins. Methods Enzymol 233:346–357CrossRefGoogle Scholar
  24. Logani MK, Davies RE (1980) Lipid oxidation: biologic effects and antioxidants—a review. Lipids 15:485–495CrossRefGoogle Scholar
  25. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein estimation with Folin–phenol reagent. J Biol Chem 193:265–278Google Scholar
  26. Maestri E, Marmiroli M, Visioli G, Marmiroli N (2010) Metal tolerance and hyperaccumulation: costs and trade-offs between traits and environment. Environ Exp Bot 68(1):1–13CrossRefGoogle Scholar
  27. Małecka 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 31:1053–1063CrossRefGoogle Scholar
  28. Malecka A, Piechalak A, Mensinger A, Hanc A, Baralkiewicz D, Tomaszewska B (2012) Antioxidative defense system in Pisum sativum rooots exposed to heavy metals (Pb, Cu, Cd, Zn). Pol J Environ Stud 21:1721–1730  Google Scholar
  29. Møller IM, Kristensen BK (2004) Protein oxidation in plant mitochondria as a stress indicator. Photochem Photobiol Sci 3(8):730–735CrossRefGoogle Scholar
  30. Nadgórska-Socha A, Kandziora-Ciupa M, Ciepał R, Walasek K (2011) Effects of Zn, Cd, Pb on physiological response of Silene vulgaris plants from selected populations. Pol J Environ Stud 20:599–604Google Scholar
  31. Nakano Y, Asada K (1987) Purification of ascorbate peroxidase in spinach chloroplasts; its inactivation in ascorbate-depleted medium and reactivation by monodehydroascorbate radical. Plant Cell Physiol 28:131–140Google Scholar
  32. Pena LB, Zawoznik MS, Tornado ML, Gallego SM (2008) Heavy metals effects on proteolytic system in sunflower leaves. Chemosphere 72:741–746CrossRefGoogle Scholar
  33. Pirslova B, Kuna R, Libantova J, Moravcikova J, Matusikova I (2011) Biochemical and physiological comparison of heavy metal-triggered defense responses in the monocot maize and dicot soybean roots. Mol Biol Rep 38:3437–3446CrossRefGoogle Scholar
  34. Pourrut B, Shahid M, Dumat C, Winterton P, Pinelli E (2011) Lead uptake, toxicity, and detoxification in plants. Rev Environ Contam Toxicol 213:113–136Google Scholar
  35. 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
  36. Rucińska-Sobkowiak R, Pukacki PM (2006) Antioxidative defense system in lupin roots exposed to increasing concentrations of lead. Acta Physiol Plant 28:357–364CrossRefGoogle Scholar
  37. Schützendūbel A, Schwanz P, Teichmann T, Gross K, Langenfeld-Heyser R, Godbold DL, Polle A (2001) Cadmium-induced changes in antioxidative systems, hydrogen peroxide content, and differentiation in Scots pine roots. Plant Physiol 127:887–898CrossRefGoogle Scholar
  38. Sengar RS, Gautam M, Sengar RS, Garg SK, Sengar K, Chaudhary R (2008) Lead stress effects on physiobiochemical activities of higher plants. Rev Environ Contam Toxicol 196:73–93Google Scholar
  39. Seregin IV, Shpigun LK, Ivanon VB (2004) Distribution and toxic effects of cadmium and lead on maize roots. Russ J Plant Physiol 51:525–533CrossRefGoogle Scholar
  40. Seth CS, Remans T, Keunen E, Jozefczak M, Gielen H, Opdenakker K, Weyens N, Vangronsveld J, Cuypers A (2012) Phytoextraction of toxic metals: a central role for glutathione. Plant Cell Environ 35(2):334–346CrossRefGoogle Scholar
  41. Sharma P, Dubey RS (2005) Lead toxicity in plants. Braz J Plant Physiol 17(1):35–52CrossRefGoogle Scholar
  42. Singh HP, Batish DR, Kaur G, Arora K, Kohli RK (2008) Nitric oxide (as sodium nitroprusside) supplementation ameliorates Cd toxicity in hydroponically grown wheat roots. Environ Exp Bot 63:158–167CrossRefGoogle Scholar
  43. Stone JR, Yang S (2006) Hydrogen peroxide: a signaling messenger. Antioxid Redox Signal 8:243–270CrossRefGoogle Scholar
  44. Thordal-Christensen H, Zhang Z, Wei Y, Collinge DB (1997) Subcellular localization of H2O2 in plants, H2O2 accumulation in papillae and hypersensitive response during barley–powdery mildew interaction. Plant J 11:1187–1194CrossRefGoogle Scholar
  45. Uzu G, Sobanska S, Sarret G, Munoz M, Dumat C (2010) Foliar lead uptake by lettuce exposed to atmospheric fallouts. Environ Sci Technol 44:1036–1042CrossRefGoogle Scholar
  46. 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

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Gurpreet Kaur
    • 1
  • Shubhpreet Kaur
    • 2
  • Harminder Pal Singh
    • 1
  • Daizy Rani Batish
    • 2
  • Ravinder Kumar Kohli
    • 2
  • Valbha Rishi
    • 3
  1. 1.Department of Environment StudiesPanjab UniversityChandigarhIndia
  2. 2.Department of BotanyPanjab UniversityChandigarhIndia
  3. 3.Chandigarh College of Engineering and TechnologyChandigarhIndia

Personalised recommendations