Tartaric Acid Mediated Cr Hyperaccumulation and Biochemical alterations in seedlings of Hordeum vulgare L.

  • Manik Sharma
  • Vinod KumarEmail author
  • Renu Bhardwaj
  • Ashwani Kumar Thukral


The present study was done to evaluate the effects of tartaric acid (TA) amendment on physiology of Hordeum vulgare L. seedlings under Cr(VI) stress. Cr(VI) at higher concentrations decreased the shoot and root dry weights of seedlings. However, amendment of Cr(VI) media with TA enhanced the root and shoot dry weights, activities of antioxidative enzymes (ascorbate peroxidase, guaiacol peroxidase, superoxide dismutase, catalase, and glutathione reductase), and contents of pigments (total chlorophyll and carotenoids). Cr(VI) also increased the malondialdehyde content, an indicator of lipid peroxidation and stress. However, application of TA in combination with higher concentrations of Cr(VI) reduced the malondialdehyde content. Amendment of 0.5 mM Cr(VI) with 1.5 mM TA increased the Cr uptake content in the roots of the seedlings by 208.7% with respect to treatment with 0.5 mM Cr(VI) only. The shoot and root bio-concentration factors (BCF) were enhanced with the application of TA to culture media. Further, the results were statistically analyzed by employing various multivariate techniques such as analysis of variance, multiple regression analysis, beta regression analysis, correlation analysis, principal component analysis, factor analysis, non-metric multidimensional scaling, canonical correspondence analysis, and two-block partial least squares. The present study confirmed that TA acts as an antagonist to Cr(VI) in the seedlings of H. vulgare by increasing their antioxidative potential and enhancing their capability of chromium accumulation. The present study also suggests that multivariate techniques, which are mainly applied for data analysis in the field of ecology, can also be applied for experimental biology studies.


Barley Cr(VI) Antioxidants Pigments Multivariate analysis 



The authors are thankful to the University Grants Commission, GOI for providing financial assistance in the form of the major research project and UGC-BSR fellowship to MS.

Compliance with Ethical Standards

Conflict of interest

The authors have declared no conflict of interest.

Supplementary material

344_2019_9959_MOESM1_ESM.docx (208 kb)
Supplementary material 1 (DOCX 207 KB)


  1. Aebi H (1984) Catalase in vitro. Methods Enzymol Acad Press 105:12–21Google Scholar
  2. Afshan S, Ali S, Bharwana SA, Rizwan M, Farid M, Abbas F, Abbasi GH (2015) Citric acid enhances the phytoextraction of chromium, plant growth, and photosynthesis by alleviating the oxidative damages in Brassica napus L. Environ Sci Pollut Res 22(15):11679–11689Google Scholar
  3. Ali S, Bharwana SA, Rizwan M, Farid M, Kanwal S, Ali Q, Khan MD (2015) Fulvic acid mediates chromium (Cr) tolerance in wheat (Triticum aestivum L.) through lowering of Cr uptake and improved antioxidant defense system. Environ Sci Pollut Res 22(14):10601–10609Google Scholar
  4. Ali J, Mahmood T, Hayat K, Afridi MS, Ali F, Chaudhary HJ (2018) Phytoextraction of Cr by maize (Zea mays L.). The role of plant growth promoting endophyte and citric acid under polluted soil. Arc Environ Prot 44:73–82Google Scholar
  5. Allen SE, Grimshaw HM, Parkinson JA, Quarmby C, Roberts JD (1976) Chemical Analysis. In: Chapman SB (ed) Methods in plant ecology. Blackwell Scientific Publications, Oxford, pp 424–426Google Scholar
  6. Arnon DI (1949) Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol 24(1):1–15Google Scholar
  7. ATSDR (Agency for Toxic Substances and Disease Registry) (1998) Toxicological profile for chromium. U.S. Public Health Service, U.S. Department of Health and Human Services, AtlantaGoogle Scholar
  8. Bala R, Thukral AK (2011) Phytoremediation of Cr (VI) by Spirodela polyrrhiza L. Schleiden employing reducing and chelating agents. Int J Phytoremediation 13:465–491Google Scholar
  9. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72(1–2):248–254Google Scholar
  10. Carlberg I, Mannervik B (1975) Purification and characterization of the flavoenzyme glutathione reductase from rat liver. J Biol Chem 250(14):5475–5480Google Scholar
  11. Choudhury S, Panda SK (2005) Toxic effects, oxidative stress and ultrastructural changes in moss Taxithelium nepalense (Schwaegr.) Broth. under chromium and lead phytotoxicity. Water Air Soil Poll 167:73–90Google Scholar
  12. Dakora FD, Phillips DA (2002) Root exudates as mediators of mineral acquisition in low-nutrient environments. Plant Soil 245(1):35–47Google Scholar
  13. Daud MK, Mei L, Variath MT, Ali S, Li C, Rafiq MT, Zhu SJ (2014) Chromium (VI) uptake and tolerance potential in cotton cultivars: effect on their root physiology, ultramorphology, and oxidative metabolism. BioMed Res Int. Google Scholar
  14. Fan X, Wen X, Huang F, Cai Y, Cai K (2016) Effects of silicon on morphology, ultrastructure and exudates of rice root under heavy metal stress. Acta Physiol Plant 38(8):197. Google Scholar
  15. Farid M, Ali S, Rizwan M, Ali Q, Abbas F, Bukhari SAH, Saeed R, Wu L (2017) Citric acid assisted phytoextraction of chromium by sunflower; morpho-physiological and biochemical alterations in plants. Ecotoxicol Environ Safe 145:90–102Google Scholar
  16. Farid M, Ali S, Rizwan M, Ali Q, Saeed R, Nasir T, Ahmad T (2018) Phyto-management of chromium contaminated soils through sunflower under exogenously applied 5-aminolevulinic acid. Ecotoxicol Environ Safe 151:255–265Google Scholar
  17. Gautam M, Agrawal M (2017) Phytoremediation of metals using vetiver (Chrysopogon zizanioides (L.) Roberty) grown under different levels of red mud sludge amended soil. J Geochem Explor 182:218–227Google Scholar
  18. Gill RA, Zang L, Ali B, Farooq MA, Cui P, Yang S, Zhou W (2015) Chromium-induced physio-chemical and ultrastructural changes in four cultivars of Brassica napus L. Chemosphere 120:154–164Google Scholar
  19. Gomes-Junior RA, Moldes CA, Delite FS, Pompeu GB, Gratao PL, Mazzafera P, Lea PG, Azevedo RA (2006) Antioxidant metabolism of coffee cell suspension cultures in response to cadmium. Chemosphere 65:1330–1337Google Scholar
  20. Govindasamy C, Arulpriya M, Ruban P, Jenifer FL, Ilayaraja A (2011) Concentration of heavy metals in seagrasses tissue of the Palk Strait, Bay of Bengal. Int J Environ Sci 2:145–153Google Scholar
  21. Habiba U, Ali S, Farid M, Shakoor MB, Rizwan M, Ibrahim M et al (2015) EDTA enhanced plant growth, antioxidant defense system, and phytoextraction of copper by Brassica napus L. Environ Sci Pollut Res 22(2):1534–1544Google Scholar
  22. Handa N, Kohli SK, Thukral AK, Arora S, Bhardwaj R (2017) Role of Se (VI) in counteracting oxidative damage in Brassica juncea L. under Cr (VI) stress. Acta Physiol Plant 39(2):51Google Scholar
  23. Heath RL, Packer L (1968) Photoperoxidation in isolated chloroplasts: I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys 125(1):189–198Google Scholar
  24. Hegedüs A, Erdei S, Horváth G (2001) Comparative studies of H2O2 detoxifying enzymes in green and greening barley seedlings under cadmium stress. Plant Sci 160(6):1085–1093Google Scholar
  25. Jabeen N, Abbas Z, Iqbal M, Rizwan M, Jabbar A, Farid M, Ali S, Ibrahim M, Abbas F (2016) Glycinebetaine mediates chromium tolerance in mung bean through lowering of Cr uptake and improved antioxidant system. Arch Agron Soil Sci 62:648–662Google Scholar
  26. Kanwar MK, Poonam, Pal S, Bhardwaj R (2015) Involvement of Asada-Halliwell pathway during phytoremediation of chromium (VI) in Brassica juncea L. plants. Int J Phytoremediation 17(12):1237–1243Google Scholar
  27. Kaur R, Yadav P, Sharma A, Thukral AK, Kumar V, Kohli SK, Bhardwaj R (2017) Castasterone and citric acid treatment restores photosynthetic attributes in Brassica juncea L. under Cd (II) toxicity. Ecotoxicol Environ Saf 145:466–475Google Scholar
  28. Kaur R, Kaur R, Sharma A, Kumar V, Sharma M, Bhardwaj R, Thukral AK (2018) Microbial production of dicarboxylic acids from edible plants and milk using GC-MS. J Anal Sci Technol 9(1):21. Google Scholar
  29. Kono Y (1978) Generation of superoxide radical during autoxidation of hydroxylamine and an assay for superoxide dismutase. Arch Biochem Biophys 186(1):189–195Google Scholar
  30. Kotaś J, Stasicka Z (2000) Chromium occurrence in the environment and methods of its speciation. Environ Pollut 107(3):263–283Google Scholar
  31. Kumar D, Tripathi DK, Chauhan DK (2014) Phytoremediation potential and nutrient status of Barringtonia acutangula Gaerth. Tree seedlings grown under different chromium (CrVI) treatments. Biol Trace Elem Res 157:164–174Google Scholar
  32. Kwak S, Yoo JC, Moon DH, Baek K (2018) Role of clay minerals on reduction of Cr (VI). Geoderma 312:1–5Google Scholar
  33. Lichtenthaler HK (1987) Chlorophylls and carotenoids: pigments of photosynthetic biomembranes. Methods Enzymol Acad Press 148:350–382Google Scholar
  34. Linero O, Cidad M, Carrero JA, Nguyen C, de Diego A (2015) Accumulation and translocation of essential and nonessential elements by tomato plants (Solanum lycopersicum) cultivated in open-air plots under organic or conventional farming techniques. J Agric Food Chem 63(43):9461–9470Google Scholar
  35. Lu LL, Tian SK, Yang XE, Peng HY, Li TQ (2013) Improved cadmium uptake and accumulation in the hyperaccumulator Sedum alfredii: the impact of citric acid and tartaric acid. J Zhejiang Univ Sci B 14(2):106–114Google Scholar
  36. Ma J, Lv C, Xu M, Chen G, Lv C, Gao Z (2016a) Photosynthesis performance, antioxidant enzymes, and ultrastructural analyses of rice seedlings under chromium stress. Environ Sci Pollut Res 23(2):1768–1778Google Scholar
  37. Ma Q, Cao X, Wu L, Mi W, Feng Y (2016b) Light intensity affects the uptake and metabolism of glycine by pakchoi (Brassica chinensis L.). Sci Rep 6:21200. Google Scholar
  38. Morel JL (1997) Bioavailability of trace elements to terrestrial plants. - Chap. 6. In: Tarradellas J, Bitton G, Rossel D (eds) Soil ecotoxicology. Lewis Publishers, Boca Raton, pp 141–176Google Scholar
  39. Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant Cell Physiol 22:867–880Google Scholar
  40. Noman A, Ali S, Naheed F, Ali Q, Farid M, Rizwan M, Irshad MK (2015) Foliar application of ascorbate enhances the physiological and biochemical attributes of maize (Zea mays L.) cultivars under drought stress. Arch Agron Soil Sci 61:1659–1672Google Scholar
  41. Pandey V, Dixit V, Shyam R (2005) Antioxidative responses in relation to growth of mustard (Brassica juncea cv. Pusa Jaikisan) plants exposed to hexavalent chromium. Chemosphere 61(1):40–47Google Scholar
  42. Pütter J (1974) Peroxidases. Methods Enzym Anal 2:685–690Google Scholar
  43. Rizwan M, Meunier JD, Miche H, Keller C (2012) Effect of silicon on reducing cadmium toxicity in durum wheat (Triticum turgidum L. cv. Claudio W.) grown in a soil with aged contamination. J Hazard Mater 209:326–334Google Scholar
  44. Rizwan M, Ali S, Adrees M, Rizvi H, Zia-ur-Rehman M, Hannan F, Qayyum MF, Hafeez F, Ok YS (2016a) Cadmium stress in rice: toxic effects, tolerance mechanisms, and management: a critical review. Environ Sci Pollut Res 23:17859–17879Google Scholar
  45. Rizwan M, Ali S, Rizvi H, Rinklebe J, Tsang DC, Meers E, Ok YS, Ishaque W (2016b) Phytomanagement of heavy metals in contaminated soils using sunflower: a review. Crit Rev Environ Sci Technol 46:1498–1528Google Scholar
  46. Rucinska-Sobkowiak R, Pukacki PM (2006) Antioxidative defense system in lupin roots exposed to increasing concentrations of lead. Acta Physiol Plant 28:357–364Google Scholar
  47. Salt DE, Blaylock M, Kumar PBAN, Duschenkov V, Ensley BD, Chet I, Raskin I (1995) Phytoremediation: a novel strategy for the removal of toxic metals from the environment using plants. Biotechnology 13:468–475Google Scholar
  48. Shahid MN, Abbasi NA (2011) Effect of beewax coatings on physiological changes in Fruits of sweet orange cv.“blood red”. Sarhad J Agric 27(3):385–394Google Scholar
  49. Shahid M, Dumat C, Silvestre J, Pinelli E (2012) Effect of fulvic acids on lead-induced oxidative stress to metal sensitive Vicia faba L. plant. Biol Fertil Soils 48(6):689–697Google Scholar
  50. Shahzad B, Tanveer M, Che Z, Rehman A, Cheema SA, Sharma A et al (2018a) Role of 24-epibrassinolide (EBL) in mediating heavy metal and pesticide induced oxidative stress in plants: a review. Ecotoxicol Environ Saf 147:935–944Google Scholar
  51. Shahzad B, Tanveer M, Rehman A, Cheema SA, Fahad S, Rehman S, Sharma A (2018b) Nickel; whether toxic or essential for plants and environment-a review. Plant Physiol Biochem 132:641–651Google Scholar
  52. Shakir L, Ejaz S, Ashraf M, Qureshi NA, Anjum AA, Iltaf I, Javeed A (2012) Ecotoxicological risks associated with tannery effluent wastewater. Environ Toxicol Pharmacol 34(2):180–191Google Scholar
  53. Shakoor MB, Ali S, Hameed A, Farid M, Hussain S, Yasmeen T, Abbasi GH (2014) Citric acid improves lead (Pb) phytoextraction in Brassica napus L. by mitigating Pb-induced morphological and biochemical damages. Ecotoxicol Environ Saf 109:38–47Google Scholar
  54. Shapiro SS, Wilk MB (1965) Analysis of variance test for normality (complete samples). Biometrika 52: 591-etr. Online version implemented by Simon Dittami (2009)Google Scholar
  55. Sharma A, Bhardwaj R, Kumar V, Thukral AK (2016a) GC-MS studies reveal stimulated pesticide detoxification by brassinolide application in Brassica juncea L. plants. Environ Sci Pollut Res 23(14):14518–14525Google Scholar
  56. Sharma A, Kumar V, Singh R, Thukral AK, Bhardwaj R (2016b) Effect of seed pre-soaking with 24-epibrassinolide on growth and photosynthetic parameters of Brassica juncea L. in imidacloprid soil. Ecotoxicol Environ Saf 133:195–201Google Scholar
  57. Sharma A, Kumar V, Thukral AK, Bhardwaj R (2016c) Epibrassinolide-imidacloprid interaction enhances non-enzymatic antioxidants in Brassica juncea L. Indian J Plant Physiol 21(1):70–75Google Scholar
  58. Sharma A, Thakur S, Kumar V, Kanwar MK, Kesavan AK, Thukral AK, Ahmad P (2016d) Pre-sowing seed treatment with 24-epibrassinolide ameliorates pesticide stress in Brassica juncea L. through the modulation of stress markers. Front Plant Sci 7:1569. Google Scholar
  59. Sharma A, Kumar V, Kanwar MK, Thukral AK, Bhardwaj R (2017a) Ameliorating imidacloprid induced oxidative stress by 24-epibrassinolide in Brassica juncea L. Russ J Plant Physiol 64(4):509–517Google Scholar
  60. Sharma A, Thakur S, Kumar V, Kesavan AK, Thukral AK, Bhardwaj R (2017b) 24-Epibrassinolide stimulates imidacloprid detoxification by modulating the gene expression of Brassica juncea L. BMC Plant Biol 17(1):56. Google Scholar
  61. Sharma A, Kumar V, Kumar R, Shahzad B, Thukral AK, Bhardwaj R (2018a) Brassinosteroid-mediated pesticide detoxification in plants: a mini-review. Cogent Food Agric 4(1):1436212Google Scholar
  62. Sharma A, Kumar V, Yuan H, Kanwar MK, Bhardwaj R, Thukral AK, Zheng B (2018b) Jasmonic acid seed treatment stimulates insecticide detoxification in Brassica juncea L. Front Plant Sci. Google Scholar
  63. Sharma R, Bhardwaj R, Gautam V, Bali S, Kaur R, Kaur P et al (2018c) Phytoremediation in Waste management: hyperaccumulation diversity and techniques. In: Plants under metal and metalloid stress. Springer, Singapore, pp 277–302Google Scholar
  64. Stambulska UY, Bayliak MM, Lushchak VI (2018) Chromium (VI) toxicity in legume plants: modulation effects of rhizobial symbiosis. BioMed Res Int 2018:1–13Google Scholar
  65. Tian HZ, Zhu CY, Gao JJ, Cheng K, Hao JM, Wang K, Zhou JR (2015) Quantitative assessment of atmospheric emissions of toxic heavy metals from anthropogenic sources in China: historical trend, spatial distribution, uncertainties, and control policies. Atmos Chem Phys 15(17):10127–10147Google Scholar
  66. Verma S, Dubey RS (2003) Lead toxicity induces lipid peroxidation and alters the activities of antioxidant enzymes in growing rice plants. Plant Sci 164(4):645–655Google Scholar
  67. Wang H, Zhong G (2011) Effect of organic ligands on accumulation of copper in hyperaccumulator and nonaccumulator Commelina communis. Biol Trace Elem Res 143(1):489–499Google Scholar
  68. Wiszniewska A, Hanus-Fajerska E, Muszyńska E, Ciarkowska K (2016) Natural organic amendments for improved phytoremediation of polluted soils: a review of recent progress. Pedosphere 26:1–12Google Scholar
  69. Yadav K, Singh NB (2013) Effects of benzoic acid and cadmium toxicity on wheat seedlings. Chil J Agric Res 73(2):168–174Google Scholar
  70. Yadav P, Kaur R, Kanwar MK, Sharma A, Verma V, Sirhindi G, Bhardwaj R (2018) Castasterone confers copper stress tolerance by regulating antioxidant enzyme responses, antioxidants, and amino acid balance in B. juncea seedlings. Ecotoxicol Environ Saf 147:725–734Google Scholar
  71. Yıldız M, Terzi H (2016) Proteomic analysis of chromium stress and sulfur deficiency responses in leaves of two canola (Brassica napus L.) cultivars differing in Cr (VI) tolerance. Ecotoxicol Environ Saf 124:255–266Google Scholar
  72. Zaheer IE, Ali S, Rizwan M, Farid M, Shakoor MB, Gill RA, Ahmad R (2015) Citric acid assisted phytoremediation of copper by Brassica napus L. Ecotoxicol Environ Saf 120:310–317Google Scholar
  73. Zou JH, Wang M, Jiang WS, Liu DH (2006) Effects of hexavalent chromium (VI) on root growth and cell division in root tip cells of Amaranthus viridis L. Pak J Bot 38(3):673Google Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Botanical and Environmental SciencesGuru Nanak Dev UniversityAmritsarIndia
  2. 2.Department of BotanyDAV UniversityJalandharIndia

Personalised recommendations