Oxidative injury and antioxidant enzymes regulation in arsenic-exposed seedlings of four Brassica napus L. cultivars

Abstract

Environmental contamination due to arsenic (As) has become a major risk throughout the world; this affects plant growth and productivity. Its accumulation in food chain may pose a severe threat to organisms. The present study was carried out to observe the toxic effects of As (0, 50, 100, and 200 μM) on physiological and biochemical changes in four Brassica napus cultivars (ZS 758, Zheda 619, ZY 50, and Zheda 622). Results showed that As toxicity provoked a significant inhibition in growth parameters of B. napus cultivars and this reduction was more obvious in cultivar Zheda 622. The highest concentration of MDA, H2O2, and O2 contents in both leaf and root tissues were observed at 200 μM As level, and a gradual decrease was observed at lower concentrations. Increasing As concentration gradually decreased chlorophyll and carotenoids contents. Activity of antioxidant enzymes such as SOD, CAT, APX, GR, and GSH was positively correlated with As treatments in all cultivars. The microscopic study of leaves and roots at 200 μM As level showed the disorganization in cell organelles. Disturbance in the morphology of chloroplast, broken cell wall, increase in size, and number of starch grains and immature nucleus were found in leaf ultrastructures under higher concentration of As. Moreover, damaged nucleus, diffused cell wall, enlarged vacuoles, and a number of mitochondria were observed in root tip cells at 200 μM As level. These results suggest that B. napus cultivars have efficient mechanism to tolerate As toxicity, as evidenced by an increased level of antioxidant enzymes.

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References

  1. Aebi H (1984) Catalase in vitro. Methods Enzymol 10:121–126

    Article  Google Scholar 

  2. Ali B, Huang CR, Qi ZY, Ali S, Daud MK, Geng XX, Liu HB, Zhou WJ (2013) 5 Aminolevulinic acid ameliorates cadmium-induced morphological, biochemical and ultrastructural changes in seedlings of oilseed rape. Environ Sci Pollut Res 20:7256–7267

    CAS  Article  Google Scholar 

  3. Ali B, Qian P, Jin R, Ali S, Khan M, Aziz R, Tian T, Zhou WJ (2014a) Physiological and ultra-structural changes in Brassica napus seedlings induced by cadmium stress. Biol Plant 58:131–138

    CAS  Article  Google Scholar 

  4. Ali B, Qian P, Sun R, Farooq MA, Gill RA, Wang J, Azam M, Zhou WJ (2014b) Hydrogen sulfide alleviates the aluminum-induced changes in Brassica napus as revealed by physio-chemical and ultrastructural study of plant. Environ Sci Pollut Res DOI:. doi:10.1007/s11356-014-3551-y

    Google Scholar 

  5. Ansari MKA, Shao HB, Umar S, Ahmad A, Ansari HS, Iqbal M, Owens G (2013) Screening Indian mustard genotypes for phytoremediating arsenic-contaminated soils. Clean–Soil Air Water 41:195–201

    CAS  Article  Google Scholar 

  6. Baker CJ, Mock NM (1994) An improved method for monitoring cell death in suspension and leaf disc assays using Evans blue. Plant Cell Tiss Org 39:7–12

    Article  Google Scholar 

  7. Bhattacharya S, Gupta K, Debnath S, Ghosh UC, Chattopadhyay DJ, Mukhopadhyay A (2012) Arsenic bioaccumulation in rice and edible plants and subsequent transmission through food chain in Bengal basin: a review of the perspectives for environmental health. Toxicol Environ Chem 94:429–441

    CAS  Article  Google Scholar 

  8. Britt AB (1999) Molecular genetics of DNA repair in higher plants. Trends Plant Sci 4:20–25

    Article  Google Scholar 

  9. Cao H, Jiang Y, Chen J, Zhang H, Huang W, Li L, Zhang W (2009) Arsenic accumulation in Scutellaria Baicalensis Georgi and its effects on plant growth and pharmaceutical components. J Hazard Mater 171:508–513

    CAS  Article  Google Scholar 

  10. Chakrabarty D, Trivedi PK, Misra P, Tiwari M, Shri M, Shukla D, Kumar S, Rai A, Pandey A, Nigam D, Tripathi RD, Tuli R (2009) Comparative transcriptome analysis of arsenate and arsenite stresses in rice seedlings. Chemosphere 74:688–702

    CAS  Article  Google Scholar 

  11. Choudhury B, Chowdhury S, Biswas AK (2011) Regulation of growth and metabolism in rice (Oryza sativa L.) by arsenic and its possible reversal by phosphate. J Plant Inter 6:15–24

    CAS  Google Scholar 

  12. Chun L, Shu F, Yun S, Li J, Yang LX, Xiao H (2007) Effects of arsenic on seed germination and physiological activities of wheat seedlings. J Environ Sci 19:725–732

    Article  Google Scholar 

  13. Czerpak R, Piotrowska A, Szleska K (2006) Jasmonic acid affects changes in the growth and some components content in alga Chlorella vulgaris. Acta Physiol Plant 28:195–203

    CAS  Article  Google Scholar 

  14. Drazkiewicz M, Tukendorf A, Baszynski T (2003) Age-dependent response of maize leaf segments to cadmium treatment: Effect on chlorophyll fluorescence and phytochelatin accumulation. J Plant Physiol 160:247–254

    CAS  Article  Google Scholar 

  15. Fatima A, Ahmad M (2004) Certain antioxidant enzymes of Allium cepa as biomarkers for the detection of toxic heavy metals in wastewater. Sci Total Environ 346:256–273

    Article  Google Scholar 

  16. Foyer CH, Noctor G (2005) Oxidant and antioxidant signaling in plants: a reevaluation of the concept of oxidative stress in a physiological context. Plant Cell Environ 28:1066–1071

    Article  Google Scholar 

  17. Fridovich I (1995) Superoxide radical and superoxide dismutases. Ann Review Bioche 64:97–112

    CAS  Article  Google Scholar 

  18. Gallego SM, Benavides MP, Tomaro ML (1996) Effect of heavy metal ion excess on sunflower leaves: evidence for involvement of oxidative stress. Plant Sci 121:151–159

    CAS  Article  Google Scholar 

  19. Geng CN, Zhu YG, Tong YP, Smith SE, Smith FA (2006) Arsenate (As) uptake by and distribution in two cultivars of winter wheat (Triticum aestivum L.). Chemosphere 62:608–615

    CAS  Article  Google Scholar 

  20. Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem 48:909–930

    CAS  Article  Google Scholar 

  21. Gill RA, Hu XQ, Ali B, Yang C, Shou JY, Wu YY, Zhou WJ (2014) Genotypic variation of the responses to chromium toxicity in four oilseed rape cultivars. Biol Plant 58:539–550

    CAS  Article  Google Scholar 

  22. Gill RA, Zang L, Ali B, Farooq MA, Cui P, Yang S, Ali S, Zhou W (2015) Chromium-induced physio-chemical and ultrastructural changes in four cultivars of Brassica napus L. Chemosphere 120:154–164

    CAS  Article  Google Scholar 

  23. Gisbert C, Clemente R, Aviño JN, Baixauli C, Giner A, Serrano R, Walker DJ, Bernal MP (2006) Tolerance and accumulation of heavy metals by Brassicaceae species grown in contaminated soils from Mediterranean regions of Spain. Environ Exp Bot 56:19–27

    CAS  Article  Google Scholar 

  24. Gratão PL, Polle A, Lea PJ, Azevedo RA (2005a) Making the life of heavy metal- stressed plants a little easier. Funct Plant Biol 32:481–494

    Article  Google Scholar 

  25. Gratão PL, Prasad MNV, Cardoso PF, Lea P, Azevedo RA (2005b) Phytoremediation: green technology for the cleanup of toxic metals in the environment. Braz J Plant Physiol 17:53–64

    Article  Google Scholar 

  26. Gupta M, Sharma P, Sarin NB, Sinha AK (2009) Differential response of arsenic stress in two varieties of Brassica juncea L. Chemosphere 74:1201–1208

    CAS  Article  Google Scholar 

  27. Heikens A (2006) Arsenic contamination of irrigation water, soil and crops in Bangladesh: risk implications for sustainable agriculture and food safety is Asia. FAO-RAP 2006/20. Food and Agricultural Organization (FAO), Rome, Italy

  28. Jiang M, Zhang J (2001) Effect of abscisic acid on active oxygen species, antioxidative defence system and oxidative damage in leaves of maize seedlings. Plant Cell Physiol 42:1265–1273

    CAS  Article  Google Scholar 

  29. Jiang M, Zhang J (2002) Water stress-induced abscisic acid accumulation triggers the increased generation of reactive oxygen species and up-regulates the activities of antioxidant enzymes in maize leaves. J Exp Bot 53:2401–2410

    CAS  Article  Google Scholar 

  30. 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 210:899–903

    CAS  Google Scholar 

  31. Liu WJ, Zhu YG, Smith FA (2005) Effects of iron and manganese plaques on arsenic uptake by rice seedlings (Oryza sativa L.) grown in solution culture supplied with arsenate and arsenite. Plant Soil 277:127–138

    CAS  Article  Google Scholar 

  32. Lombi E, Zhao F-J, Fuhrmann M, Ma SQ, Mcgrath SP (2002) Arsenic distribution and speciation in the fronds of the hyper accumulator Pteris vittata. New Phytol 156:195–203

    CAS  Article  Google Scholar 

  33. Meng H, Hua S, Shamsi HI, Jilani G, Li Y, Jiang L (2009) Cadmium-induced stress on the seed germination and seedling growth of Brassica napus L., and its alleviation through exogenous plant growth regulators. Plant Growth Regul 58:47–59

    CAS  Article  Google Scholar 

  34. Momoh EJJ, Zhou WJ, Kristiansson B (2002) Variation in the development of secondary dormancy in oilseed rape genotypes under conditions of stress. Weed Res 42:446–455

    Article  Google Scholar 

  35. Mumthas S, Chidambaram AA, Sundaramoorthy P, Ganesh KS (2010) Effect of arsenic and manganese on root growth and cell division in root tip cells of green gram (Vigna radiata L.). J Food Agric 22:285–297

    Article  Google Scholar 

  36. Nakano Y, Asada K (1981) Hydrogen-peroxide is scavenged by ascorbate-specific peroxidase in spinach-chloroplasts. Plant Cell Physiol 22:867–880

    CAS  Google Scholar 

  37. Panda SK (2007) Chromium-mediated oxidative stress and ultrastructural changes in root cells of developing rice seedlings. J Plant Physiol 164:1419–1428

    CAS  Article  Google Scholar 

  38. Papoyan A, Pineros M, Kochian LV (2007) Plant Cd2+ and Zn2+ status effects on root and shoot heavy metal accumulation in Thlaspi caerulescens. New Phytol 175:51–58

    CAS  Article  Google Scholar 

  39. Pietrini F, Iannelli MA, Pasqualini S, Massacci A (2003) Interaction of cadmium with glutathione and photosynthesis in developing leaves and chloroplasts of Phragmites australis (Cav.) Trin. ex Steudel. Plant Physiol 133:829–837

    CAS  Article  Google Scholar 

  40. Poonam KR, Bali S, Singh R, Pati PK, Bhardwaj R (2014) Treatment of 24-EBL to Brassica juncea plants under Cu-metal stress lowers oxidative burst by activity of antioxidative enzymes. J Stress Physiol Biochem 10:315–327

    Google Scholar 

  41. Porra RJ, Thompson WA, Kriedemann PE (1989) Determination of accurate extinction coefficients and simultaneous equations for assaying chlorophylls a and b extracted with four different solvents: verification of the concentration of chlorophyll standards by atomic absorption spectroscopy. Biochim Biophys Acta 975:384–394

    CAS  Article  Google Scholar 

  42. Quaghebeur M, Rengel Z (2004) Arsenic uptake, translocation and speciation in pho1 and pho2 mutants of Arabidopsis thaliana. Physiol Plant 120:280–286

    CAS  Article  Google Scholar 

  43. Quan LJ, Zhang B, Shi WW, Li HY (2008) Hydrogen peroxide in plants: a versatile molecule of the reactive oxygen species network. J Integrat Plant Biol 50:2–18

    CAS  Article  Google Scholar 

  44. 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

    CAS  Article  Google Scholar 

  45. Romero-Puertas MC, Rodriguez-Serrano M, Corpas FJ, Gomez M, del Rio LA, Sandalio LM (2004) Cadmium-induced subcellular accumulation of O2•− and H2O2 in pea leaves. Plant Cell Environ 27:1122–1134

  46. Sandalio LM, Dalurzo HC, Gòmez M, Romero-Puertas MC, del Río LA (2001) Cadmium-induced changes in the growth and oxidative metabolism of pea plants. J Exp Bot 52:2115–2126

    CAS  Google Scholar 

  47. Shri M, Kumar S, Chakrabarty D, Trivedi PK, Mallick S, Misra P, Shukla D, Mishra S, Srivastava S, Tripathi RD, Tuli R (2009) Effect of arsenic on growth, oxidative stress, and antioxidant system in rice seedlings. Ecotox Environ Safe 72:1102–1110

    CAS  Article  Google Scholar 

  48. Singh N, Ma LQ, Srivastava M, Rathinasabapathi B (2006) Metabolic adaptations to arsenic-induced oxidative stress in Pteris vittata L. and Pteris ensiformis L. Plant Sci 170:274–282

    CAS  Article  Google Scholar 

  49. Singh HP, Daizy BR, Kohli RK, Arora K (2007a) Arsenic-induced root growth inhibition in mung bean (Phaseolus aureus Roxb.) is due to oxidative stress resulting from enhanced lipid peroxidation. Plant Growth Regul 53:65–73

    CAS  Article  Google Scholar 

  50. Singh SK, Juwarkar AA, Kumar S, Meshram J, Fa M (2007b) Effect of amendment on Phytoextraction of arsenic by Vetiveria zizanioides from soil. Int J Environ Sci Technol 4:339–344

    CAS  Article  Google Scholar 

  51. Singh SS, Khan NA, Rahat N, Anjum NA (2008) Photosynthetic traits and activities of antioxidant enzymes in blackgram (Vigna mungo L. Hepper) under cadmium stress. Am J Plant Physiol 3:25–32

    CAS  Article  Google Scholar 

  52. Smith SE, Christophersen HM, Pope S, Smith A (2010) Arsenic uptake and toxicity in plants: integrating mycorrhizal influences. Plant Soil 327:1–21

    CAS  Article  Google Scholar 

  53. Srivastava M, Ma LQ, Singh N, Singh S (2005) Antioxidant responses of hyperaccumulator and sensitive Fern species to arsenic. Environ Exp Bot 56:1335–1342

    CAS  Article  Google Scholar 

  54. Stone JR, Yang S (2006) Hydrogen peroxide: a signaling messenger. Antioxid Redox Signal 8:243–70

    CAS  Article  Google Scholar 

  55. Sun L, Yan X, Liao X, Wen Y, Chong Z, Liang T (2011) Interactions of arsenic and phenanthrene on their uptake and antioxidative response in Pteris vittata L. Environ Pollut 159:3398–3405

    CAS  Article  Google Scholar 

  56. Thordal-Christensen H, Zhang Z, Wei YD, Collinge DB (1997) Subcellular localization of H2O2 in plants. H2O2 accumulation in papillae and hypersensitive response during the barley-powdery mildew reaction. Plant J 11:1187–1194

    CAS  Article  Google Scholar 

  57. Tripathi RD, Srivastava S, Mishra S, Singh N, Tuli R, Gupta DK, Maathuis FJM (2007) Arsenic hazards: strategies for tolerance and remediation by plants. Trends Biotech 25:158–165

    CAS  Article  Google Scholar 

  58. Velikova V, Yordanov I, Edreva A (2000) Oxidative stress and some antioxidant systems in acid rain-treated bean plants. Plant Sci 151:59–66

    CAS  Article  Google Scholar 

  59. Yadav SK (2010) Heavy metals toxicity in plants: an overview on the role of glutathione and phytochelatins in heavy metal stress tolerance of plants. S Afr J Bot 76:167–179

    CAS  Article  Google Scholar 

  60. Zhang WF, Zhang F, Raziuddin R, Gong HJ, Yang ZM, Lu L, Ye QF, Zhou WJ (2008) Effects of 5-aminolevulinic acid on oilseed rape seedling growth under herbicide toxicity stress. J Plant Growth Regul 27:159–169

    Article  Google Scholar 

  61. Zhao FJ, Wang JR, Barker JHA, Schat H, Bleeker PM, McGrath SP (2003) The role of phytochelatins in arsenic tolerance in the hyperaccumulator Pteris vittata. New Phytol 159:403–410

    CAS  Article  Google Scholar 

  62. Zhao FJ, McGrath SP, Meharg AA (2010) Arsenic as a food chain contaminant: mechanisms of plant uptake and metabolism and mitigation strategies. Ann Rev Plant Biol 61:535–559

    CAS  Article  Google Scholar 

  63. Zhou WJ, Leul M (1999) Uniconazole-induced tolerance of rape plants to heat stress in relation to changes in hormonal levels, enzyme activities and lipid peroxidation. Plant Growth Regul 27:99–104

    CAS  Article  Google Scholar 

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Correspondence to Weijun Zhou.

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Farooq, M.A., Li, L., Ali, B. et al. Oxidative injury and antioxidant enzymes regulation in arsenic-exposed seedlings of four Brassica napus L. cultivars. Environ Sci Pollut Res 22, 10699–10712 (2015). https://doi.org/10.1007/s11356-015-4269-1

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Keywords

  • Antioxidant enzyme activities
  • Arsenic stress
  • Oilseed rape
  • Microscopic study
  • Oxidative stress
  • Plasma membrane integrity