Physiology and Molecular Biology of Plants

, Volume 20, Issue 4, pp 449–460 | Cite as

Brassinosteroid-mediated evaluation of antioxidant system and nitrogen metabolism in two contrasting cultivars of Vigna radiata under different levels of nickel

  • Mohammad Yusuf
  • Qazi FariduddinEmail author
  • Iqbal Ahmad
  • Aqil Ahmad
Research Article


The role of 28-homobrassinolide (HBL) in countering nickel-induced oxidative damage through overexpression of antioxidant enzymes and proline in Vigna radiata has been investigated. Two varieties of V. radiata, one sensitive to Ni (PDM-139) and the other tolerant to Ni (T-44), were sown in the soil fed with different levels (0, 50, 100 or 150 mg kg−1) of Ni, and at 29-day stage, foliage of plants was applied with deionized water (control), 10−8 or 10−6 M of HBL. The plants were sampled at 45-day stage of growth to assess various physiological as well as biochemical characteristics. The remaining plants were allowed to grow up to maturity to study the yield characteristics. The growth traits, leghemoglobin, nitrogen and carbohydrate content in the nodules, leaf chlorophyll content, photosynthesis efficiency, leaf water potential, activities of nitrate reductase, carbonic anhydrase and nitrogenase decreased proportionately with the increasing concentrations of nickel, whereas electrolyte leakage, various antioxidant enzymes viz. catalase, peroxidase and superoxide dismutase and accumulation of proline increased at 45-day stage. However, the exogenously applied HBL to the nickel-stressed or non-stressed plants improved growth, nodulation and photosynthesis and further enhanced the various antioxidant enzymes viz. catalase, peroxidase and superoxide dismutase and accumulation of proline. The deleterious impact of Ni on the plants was concentration dependent where HBL applied to the foliage induced overexpression of antioxidant enzyme and accumulation of proline (osmolyte) which could have conferred tolerance to Ni up to 100 mg kg−1, resulting in improved growth, nodulation, photosynthesis and yield attributes.


Brassinosteroids Nickel Nitrogen metabolism Oxidative stress Proline 



M. Yusuf gratefully acknowledges the financial assistance rendered by the Science and Engineering Research Board (SERB), Department of Science and Technology (DST), New Delhi, India, in a form of Young Scientists (SB/FT/LS-210-2012).


  1. Ahmad E, Zaidi A, Khan MS, Oves M (2012) Heavy metal toxicity to symbiotic nitrogen fixing microorganism and host legumes In: Zaidi A, Wani PA, Khan MS (eds) Toxicity of heavy metals to legumes and bioremediation. Springer-VerlagGoogle Scholar
  2. Ashraf M, Akram NA, Arteca RN, Foolad MR (2010) The physiological, biochemical and molecular roles of brassinosteroids and salicylic acid in plant processes and salt tolerance. Crit Rev Plant Sci 29:162–190CrossRefGoogle Scholar
  3. Bajguz A (2000) Effect of brassinosteroids on nucleic acid and protein content in cultured cell of Chlorella vulgaris. Plant Physiol Biochem 38:209–215CrossRefGoogle Scholar
  4. Bajguz A, Asami T (2005) Suppression of Wolffiaarrhiza growth by brassinazole, an inhibitor of brassinosteroid biosynthesis and its restoration by endogenous 24-epibrassinolide. Phytochemistry 66:1787–1796PubMedCrossRefGoogle Scholar
  5. Bajguz A, Hayat S (2009) Effects of brassinosteroids on the plant responses to environmental stresses. Plant Physiol Biochem 47:1–8PubMedCrossRefGoogle Scholar
  6. Barbafieri M, Tassi E (2011) Brassinosteroids for phytoremediation application. In: Hayat S, Ahmad A (eds) Brassinosteroids: a class of plant hormone. Springer, Dordrecht, pp 403–438Google Scholar
  7. Barker AV (2006) Nickel. In: Barker AV, Pilbeam DJ (eds) Handbook of plant nutrition. CRC PressGoogle Scholar
  8. Bates LS, Walden RT, Tearse ID (1973) Rapid determination of free proline for water stress studies. Plant Soil 39:205–207CrossRefGoogle Scholar
  9. Beauchamp LO, Fridovich I (1971) Superoxide dismutase improved assays and assay applicable to acrylamide gels. Ann Biochem 44:276–287CrossRefGoogle Scholar
  10. Carpentier R (2001) The negative action of toxic divalent cations on the photosynthetic apparatus. In: Passarakli M (ed) Handbook of plant and crop physiology. Marcel Dekker, New York, pp 763–772Google Scholar
  11. Chan PK, Gresshoff PM (2009) Roles of plant hormones in legume nodulation. In: Horst W, Doelle Edgar J, DaSilva (eds) Encyclopedia of Life Support Systems (EOLSS): biotechnology. EOLSS Publishers, OxfordGoogle Scholar
  12. Chance B, Maehly AC (1956) Assay of catalase and peroxidase. Methods Enzymol 2:764–775CrossRefGoogle Scholar
  13. Chen C, Huang D, Liu J (2009) Functions and toxicity of nickel in plants: recent advances and future prospects. Clean 37:304–313Google Scholar
  14. Del Rio LA, Corpas J, Sandalio LM, Palma JM, Barroso JB (2003) Plant peroxisomes, reactive oxygen metabolism and nitric oxide. Inter Union Biochem Mol Biol (IUBMB) Life 55:71–81CrossRefGoogle Scholar
  15. Divi UK, Rahman T, Krishna P (2010) Brassinosteroid-mediated stress tolerance in Arabidopsis shows interactions with abscisic acid, ethylene and salicylic acid pathways. BMC Plant Biol 10:151PubMedCrossRefPubMedCentralGoogle Scholar
  16. Dubois M, Gills K, Hamilton JK, Robbers PA, Smith F (1956) Chlorometric method for determination of sugars and related substances. Ann Chem 28:350–356CrossRefGoogle Scholar
  17. Dwivedi RS, Randhawa NS (1974) Evolution of a rapid test of the hidden hunger of zinc in plants. Plant Soil 40:445–451CrossRefGoogle Scholar
  18. Fariduddin Q, Ahmad A, Hayat S (2004) Responses of Vigna radiata to foliar application of 28-homobrassinolide and kinetin. Biol Planta 48:465–468CrossRefGoogle Scholar
  19. Fariduddin Q, Yusuf M, Chalkoo S, Hayat S, Ahmad A (2011) 28-homobrassinolide improves growth and photosynthesis in Cucumis sativus L. through an enhanced antioxidant system in the presence of chilling stress. Photosynthetica 49:55–64CrossRefGoogle Scholar
  20. Finkelstein R, Reeves W, Ariizumi T, Steber C (2008) Molecular aspects of seed dormancy. Annu Rev Plant Biol 59:387–415PubMedCrossRefGoogle Scholar
  21. Friedlova M (2010) The influence of heavy metals on soil biological and chemical properties. Soil Water Res 5:21–27Google Scholar
  22. Gajewska E, Sklodowska M (2007) Effect of nickel on ROS content and antioxidative enzyme activities in wheat leaves. Biometals 20:27–36PubMedCrossRefGoogle Scholar
  23. Gajewska E, Sklodowska M, Slaba M, Mazur J (2006) Effect of nickel on antioxidative enzyme activities, proline and chlorophyll content in wheat shoots. Biol Planta 50:653–659CrossRefGoogle Scholar
  24. Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem 48:909–930PubMedCrossRefGoogle Scholar
  25. Gomes MMA, Ferraz TM, Netto AT, Rosa RCC, Campostrini E, Leal NR, Zullo MAT, Nunez-Vazquez M (2003) Efeitos da aplicacao de brassinosteroidesnastrocasgasosas e fluorescencia da clorofilaemmaracujazeiroamarelosubmetido a deficienciahıdrica. Braz J Plant Physiol 15:348CrossRefGoogle Scholar
  26. Gomes JRA, Moldes CA, Delite FS, Pompeu GB, Gratas PL, Mazzafera P, Lea PJ, Azevedo RA (2006) Antoxidant metabolism of coffee cell suspension cultures in response to cadmium. Chemosphere 65:1330–1337CrossRefGoogle Scholar
  27. Gudesblat GE, Russinova E (2011) Plants grow on brassinosteroids. Curr Opin Plant Biol 14:530–537PubMedCrossRefGoogle Scholar
  28. Hopkins WJ (1995) Physiology of plants under stress. In: Hopkins WJ (ed) Introduction to plant physiology. Wiley, New York, p 438Google Scholar
  29. Iwahori S, Tominaga S, Higuchi S (1990) Retardation of abscission in citrus leaf and fruitlet explants by brassinolide. Plant Growth Regul 9:119–125CrossRefGoogle Scholar
  30. Jaworski EG (1971) Nitrate reductase assay in intact plant tissue. Biochem Biophys Res Commun 43:1274–1279PubMedCrossRefGoogle Scholar
  31. Jocsak I, Vegvari GY, Droppa M (2005) Heavy metal detoxification by organic acids in barley seedlings. Acta Biol Szeged 49:99–101Google Scholar
  32. Kupper H, Kupper F, Spiller M (1998) In situ detection of heavy metal substituted chlorophylls in water plants. Photo Res 58:123–133CrossRefGoogle Scholar
  33. Lindner RC (1944) Rapid analytical method for some more common inorganic constituents of plant tissue. Plant Physiol 19:76–89PubMedCrossRefPubMedCentralGoogle Scholar
  34. Llamas A, Ullrich CI, Sanz A (2008) Ni2+ toxicity in rice: effect on membrane functionality and plant water content. Plant Physiol Biochem 46:905–910PubMedCrossRefGoogle Scholar
  35. Marschner H (2005) Mineral nutrition of higher plants mineral nutrition of higher plants. Mineral nutrition of higher plants, 6th edn. Academic, LondonGoogle Scholar
  36. Nassar NMA (2004) Polyploidy, chimera and fertility of interspecific cassava (Manihot esculenta Crantz) hybrids. Ind J Genet Plant Breed 64:132–133Google Scholar
  37. Nemhauser JL, Hong F, Chory J (2006) Different plant hormones regulate similar processes through largely non-overlapping transcriptional responses. Cell 126:467–475PubMedCrossRefGoogle Scholar
  38. Noriega GO, Balestrasse KB, Batlle A, Tomaro ML (2007) Cadmium induced oxidative stress in soybean plants also by the accumulation of δ-aminolevulinic acid. Biometals 20:841–851PubMedCrossRefGoogle Scholar
  39. Ozdemir F, Bor M, Demiral T, Turkan I (2004) Effects of 24-epibrassinolide on seed germination, seedling growth, lipid peroxidation, proline content and antioxidative system of rice (Oryza sativa L.) under salinity stress. Plant Growth Regul 42:203–211CrossRefGoogle Scholar
  40. Rahman H, Sarbreen S, Alam S, Kawai S (2005) Effect of nickel on growth and composition of metal micronutrient in barely plants grown in nutrient solution. J Plant Nutr 28:393–404CrossRefGoogle Scholar
  41. Roitsch T (1999) Source-sink regulation by sugar and stress. Curr Opin Plant Biol 2:198–206PubMedCrossRefGoogle Scholar
  42. Sadasivum S, Manickam A (1992) Biochemical methods. New Age International Pvt. Ltd. Publishers, New DelhiGoogle Scholar
  43. Sharma P, Dubey RS (2005) Modulation of nitrate reductase activity in rice seedlings under aluminium toxicity and water stress: role of osmolytes as enzyme protectant. J Plant Physiol 162:854–864PubMedCrossRefGoogle Scholar
  44. Simon T (1999) The effect of increasing rates of nickel and arsenic on the growth of radish and soil micro-flora. Rostlinna Vysoba 45:421–430 (in Czech)Google Scholar
  45. Singh K, Pandey SN (2011) Effect of nickel stress on uptake, pigments and antioxidative responses of water lettuce, Pistia stratiotes L. J Environ Biol 32:391–394PubMedGoogle Scholar
  46. Sinha S, Pandey K (2003) Nickel induced toxic effects and bioaccumulation in the submerged plant, Hydrill averticillata (L.F.) Royle under repeated metal exposure. Bull Environ Contam Toxicol 71:1175–1183PubMedCrossRefGoogle Scholar
  47. Sresty TVS, Rao KV (1999) Ultrastuctural alterations in response to zinc and nickel stress in the root cells of pigeon pea. Environ Exp Bot 41:3–13CrossRefGoogle Scholar
  48. Sullivan CY, Ross WM (1979) Selecting the drought and heart resistance in grain sorghum. In: Mussel H, Staples RC (eds) Stress physiology in crop plants. Wiley, New York, pp 263–328Google Scholar
  49. Suzuki A, Akune M, Kogiso M, Imagama Y, Osuki K, Uchiumi T, Higashi S, Han SY, Yoshida S, Asami T, Abe M (2004) Control of nodule number by the phytohormones abscisic acid in the roots of two leguminous species. Plant Cell Physiol 45:914–922PubMedCrossRefGoogle Scholar
  50. Szabados L, Savoure A (2009) Proline: a multifunctional amino acid. Trends Plant Sci 15:89–97PubMedCrossRefGoogle Scholar
  51. Tripathy BC, Bhatia B, Mohanty P (1981) Inactivation of chloroplast photosynthetic electron-transport activity by Ni2+. Biochem Biophys Acta 638:217–224Google Scholar
  52. Xia XJ, Huang LF, Zhou YH, Mao WH, Shi K, Wu JX, Asami T, Chen Z, Yu JQ (2009) Brassinosteroids promote photosynthesis and growth by enhancing activation of Rubisco and expression of photosynthetic genes in Cucumis sativus. Planta 230:1185–1196PubMedCrossRefGoogle Scholar
  53. Xia XJ, Zhou YH, Ding J, Shi K, Asami T, Chen Z, Yu JQ (2011) Induction of systemic stress tolerance by brassinosteroid in Cucumis sativus. New Phytol 191:706–720PubMedCrossRefGoogle Scholar
  54. Yih RY, Clark HE (1965) Carbohydrate and protein content of boron deficient tomato root tips in relation to anatomy growth. Plant Physiol 40:312–315PubMedCrossRefPubMedCentralGoogle Scholar
  55. Yusuf M (2011) Effect of brassinosteroids on nickel induced changes in Vigna radiata. PhD thesis, Aligarh Muslim University, Aligarh, IndiaGoogle Scholar
  56. Yusuf M, Fariduddin Q, Hayat S, Hasan SA, Ahmad A (2011) Protective responses of 28-homobrssinolide in cultivars of Triticum aestivum with different levels of nickel. Arch Environ Contam Toxicol 60:68–76PubMedCrossRefGoogle Scholar
  57. Yusuf M, Fariduddin Q, Ahmad A (2012) 24-Epibrassinolide modulates growth, nodulation, antioxidant system, and osmolyte in tolerant and sensitive varieties of Vigna radiata under different levels of nickel: a shotgun approach. Plant Physiol Biochem 57:143–153PubMedCrossRefGoogle Scholar

Copyright information

© Prof. H.S. Srivastava Foundation for Science and Society 2014

Authors and Affiliations

  • Mohammad Yusuf
    • 2
  • Qazi Fariduddin
    • 2
    Email author
  • Iqbal Ahmad
    • 1
  • Aqil Ahmad
    • 2
  1. 1.Department of Agricultural Microbiology, Faculty of Agricultural SciencesAligarh Muslim UniversityAligarhIndia
  2. 2.Plant Physiology Section, Department of Botany, Faculty of Life SciencesAligarh Muslim UniversityAligarhIndia

Personalised recommendations