Environmental Science and Pollution Research

, Volume 26, Issue 10, pp 10343–10353 | Cite as

Effect of ethylenediaminetetraacetic acid and biochar on Cu accumulation and subcellular partitioning in Amaranthus retroflexus L.

  • Na Liu
  • Jiulan Dai
  • Haoqi Tian
  • Huan He
  • Yuen ZhuEmail author
Research Article


Phytoremediation combined with amendments and stabilization technologies are two crucial methods to deal with soil contaminated with heavy metals. Copper (Cu) contamination in soil near Cu mines poses a serious threat to ecosystems and human health. This study investigated the effect of ethylenediaminetetraacetic acid (EDTA) and biochar (BC) on the accumulation and subcellular distribution of Cu in Amaranthus retroflexus L. to demonstrate the remediation mechanism of EDTA and BC at the cellular level. The role of calcium (Ca) in response to Cu stress in A. retroflexus was also elucidated. We designed a pot experiment with a randomized block of four Cu levels (0, 100, 200, 400 mg kg−1) and three treatments (control, amendment with EDTA, and amendment with BC). The subcellular components were divided into three parts (cell walls, organelles, and soluble fraction) by differential centrifugation. The results showed that EDTA amendment significantly increased (p < 0.05) the concentrations of Cu in root cell walls and all subcellular components of stems and leaves (cell walls, organelles, and the soluble fraction). EDTA amendment significantly increased (p < 0.05) the proportion of exchangeable fraction and carbonate fraction in the soil. While BC amendment significantly decreased (p < 0.05) the concentrations of Cu in root cell walls and the root soluble fraction, it had no significant effects on Cu concentrations in the subcellular components of stems and leaves. The results revealed that EDTA mainly promoted the transfer of Cu to aboveground parts and accumulation in subcellular components of stems and leaves, while BC mainly limited Cu accumulation in root cell walls and the root soluble fraction. Ca concentrations in cell walls of roots, stems, and leaves increased as the Cu stress increased in all treatment groups, indicating that Ca plays an important role in relieving Cu toxicity in Amaranthus retroflexus L.


Cu Ca Ethylenediaminetetraacetic acid (EDTA) Biochar Subcellular distribution Amaranthus retroflexus L. 


Funding information

This work was supported by the National Key R&D Program of China (2017YFD0801300); Key R&D Program of Shanxi Province of China (201703D211014); Open Foundation of Key Laboratory of Industrial Ecology and Environmental Engineering, MOE (KLIEEE-16-03); Shandong Provincial Key Research and Development Program (2016CYJS05A02); and Key Research and Development Program of Shandong (2018GSF117024)

Supplementary material

11356_2019_4448_MOESM1_ESM.doc (422 kb)
ESM 1 (DOC 421 kb)


  1. Aguilar R, Hormazábal C, Gaete H, Neaman A (2011) Spatial distribution of copper, organic matter and pH in agricultural soils affected by mining activities. J Soil Sci Plant Nut 11:125–145Google Scholar
  2. Ahmad M, Lee SS, Yang JE, Ro H, Lee YH, OK YS (2012) Effects of soil dilution and amendments (mussel shell, cow bone, and biochar) on Pb availability and phytotoxicity in military shooting range soil. Ecotox Environ Safe 79:225–231CrossRefGoogle Scholar
  3. Beesley L, Marmiroli M (2011) The immobilisation and retention of soluble arsenic, cadmium and zinc by biochar. Environ Pollut 159:474–480CrossRefGoogle Scholar
  4. Bell PF, McLaughlin MJ, Cozens G, Stevens DP, Owens G, South H (2003) Plant uptake of 14C-EDTA, 14C-citrate and 14C-histidine from chelator-buffered and conventional hydroponic solutions. Plant Soil 253:311–319CrossRefGoogle Scholar
  5. Bhatia NP, Walsh KB, Baker AJM (2005) Detection and quantification of ligands involved in nickel detoxification in a herbaceous Ni hyperaccumulator Stackhousia tryonii Bailey. J Exp Bot 56:1343–1349CrossRefGoogle Scholar
  6. Blokhina O, Virolainen E, Fagerstedt KV (2003) Antioxidants, oxidative damage and oxygen deprivation stress: a review. Ann Bot-London 91:179–194CrossRefGoogle Scholar
  7. Bolan N, Kunhikrishnan A, Thangarajan R, Kumpiene J, Park J, Makino T, Kirkham MB, Scheckel K (2014) Remediation of heavy metal (loid)s contaminated soils-to mobilize or to immobilize? J Hazard Mater 266:141–166CrossRefGoogle Scholar
  8. Brune A, Dietz K (1995) A comparative analysis of element composition of roots and leaves of barley seedlings grown in the presence of toxic cadmium, molybdenum, nickel, and zinc concentrations. J Plant Nutr 18:853–868CrossRefGoogle Scholar
  9. Buss W, Kammann C, Koyro H (2012) Biochar reduces copper toxicity in Chenopodium quinoa Willd. in a sandy soil. J Environ Qual 41:1157–1165CrossRefGoogle Scholar
  10. Cao XD, Dermatas D, Xu XF, Shen G (2008) Immobilization of lead in shooting range soils by means of cement, quicklime, and phosphate amendments. Environ Sci Pollut Res 15:120–127CrossRefGoogle Scholar
  11. Cao XD, Ma LQ, Gao B (2009) Dairy-manure derived biochar effectively sorbs lead and atrazine. Environ Sci Technol 43:3285–3291CrossRefGoogle Scholar
  12. Cataldo S, Gianguzza A, Pettignano A, Pettignano A, Sammartano S (2012) Complex formation of copper(II) and cadmium(II) with pectin and polygalacturonic acid in aqueous solution. An ISE-H+ and ISE-Me2+ electrochemical study. Int J Electrochem Sci 7:6722–6737Google Scholar
  13. Cay S, Uyanik A, Engin MS, Kutbay HG (2015) Effect of EDTA and tannic acid on the removal of cd, Ni, Pb and cu from artificially contaminated soil by Althaea rosea Cavan. Int J Phytoremediat 17:568–574CrossRefGoogle Scholar
  14. Chen Y, Li X, Shen Z (2004) Leaching and uptake of heavy metals by ten different species of plants during an EDTA-assisted phytoextraction process. Chemosphere 57:187–196CrossRefGoogle Scholar
  15. Cobbett C (2003) Heavy metals and plants-model systems and hyperaccumulators. New Phytol 159:289–293CrossRefGoogle Scholar
  16. Del Río M, Font R, Almela C, Vélez D, Montoro R, Bailón ADH (2002) Heavy metals and arsenic uptake by wild vegetation in the Guadiamar river area after the toxic spill of the Aznalcóllar mine. J Biotechnol 98:125–137CrossRefGoogle Scholar
  17. Di Palma L, Ferrantelli P, Merli C, Biancifiori F (2003) Recovery of EDTA and metal precipitation from soil flushing solutions. J Hazard Mater 103:153–168CrossRefGoogle Scholar
  18. Escrig I, Morell I (1998) Effect of calcium on the soil adsorption of cadmium and zinc in some Spanish sandy soils. Water Air Soil Pollut 105:507–520CrossRefGoogle Scholar
  19. He SY, Wu QL, He ZL (2013) Effect of DA-6 and EDTA alone or in combination on uptake, subcellular distribution and chemical form of Pb in Lolium perenne. Chemosphere 93:2782–2788CrossRefGoogle Scholar
  20. Hirschi KD (1999) Expression of Arabidopsis CAX1 in tobacco: altered calcium homeostasis and increased stress sensitivity. Plant Cell 11:2113–2122CrossRefGoogle Scholar
  21. Huang JH, Hsu SH, Wang SL (2011) Effects of rice straw ash amendment on cu solubility and distribution in flooded rice paddy soils. J Hazard Mater 186:1801–1807CrossRefGoogle Scholar
  22. Huang L, Zhang HQ, Song YY, Yang YR, Chen H, Tang M (2017) Subcellular compartmentalization and chemical forms of lead participate in lead tolerance of Robinia pseudoacacia L. with Funneliformis mosseae. Front Plant Sci 8:517–528Google Scholar
  23. Hulicova-Jurcakova D, Seredych M, Lu GQ, Bandosz TJ (2009) Combined effect of nitrogen- and oxygen-containing functional groups of microporous activated carbon on its electrochemical performance in supercapacitors. Adv Funct Mater 19:438–447CrossRefGoogle Scholar
  24. Janoušková M, Pavlíková D (2010) Cadmium immobilization in the rhizosphere of arbuscular mycorrhizal plants by the fungal extraradical mycelium. Plant Soil 332:511–520CrossRefGoogle Scholar
  25. Jiang XJ, Luo YM, Zhao QG (2001) Study on phytoremediation of cadmium contaminated soil and the EDTA regulation. Soil 33:197–201 (in Chinese)Google Scholar
  26. Jiang J, Xu RK, Jiang TY, Li Z (2012) Immobilization of cu(II), Pb(II) and cd(II) by the addition of rice straw derived biochar to a simulated polluted Ultisol. J Hazard Mater 229-230:145–150CrossRefGoogle Scholar
  27. Karami N, Clemente R, Moreno-Jiménez E, Lepp NW, Beesley L (2011) Efficiency of green waste compost and biochar soil amendments for reducing lead and copper mobility and uptake to ryegrass. J Hazard Mater 191:41–48CrossRefGoogle Scholar
  28. Kotrba P, Najmanova J, Macek T, Ruml T, Mackova M (2009) Genetically modified plants in phytoremediation of heavy metal and metalloid soil and sediment pollution. Biotechnol Adv 27:799–810CrossRefGoogle Scholar
  29. Lin YF, Aarts MG (2012) The molecular mechanism of zinc and cadmium stress response in plants. Cell Mol Life Sci 69:3187–3206CrossRefGoogle Scholar
  30. Liu N, Miao YJ, Zhou XX, Gan YD, Liu SW, Wang WX, Dai JL (2018) Roles of rhizospheric organic acids and microorganisms in mercury accumulation and translocation to different winter wheat cultivars. Agric Ecosyst Environ 258:104–112CrossRefGoogle Scholar
  31. Lu KP, Yang X, Shen JJ, Robinson B, Huang HG, Liu D, Bolan N, Pei JC, Wang HL (2014) Effect of bamboo and rice straw biochars on the bioavailability of cd, cu, Pb and Zn to Sedum plumbizincicola. Agric Ecosyst Environ 191:124–132CrossRefGoogle Scholar
  32. Ma JH, Xing GF, Yang WX, Ma LL, Gao M, Wang YG, Han YH (2012) Inhibitory effects of leachate from Eupatorium adenophorum on germination and growth of Amaranthus retroflexus and Chenopodium glaucum. Acta Ecol Sin 32:50–56CrossRefGoogle Scholar
  33. McGrath SP, Zhao FJ (2003) Phytoextraction of metals and metalloids from contaminated soils. Curr Opin Biotechnol 14:277–282CrossRefGoogle Scholar
  34. Meier S, Borie F, Bolan N, Cornejo P (2012) Phytoremediation of metal-polluted soils by arbuscular mycorrhizal fungi. Crit Rev Environ Sci Technol 42:741–775CrossRefGoogle Scholar
  35. Meier S, Curaqueo G, Khan N, Bolan N, Cea M (2017) Chicken-manure-derived biochar reduced bioavailability of copper in a contaminated soil. J Soils Sediments 17:741–750CrossRefGoogle Scholar
  36. Mühlbachová G (2011) Soil microbial activities and heavy metal mobility in long-term contaminated soils after addition of EDTA and EDDS. Ecol Eng 37:1064–1071CrossRefGoogle Scholar
  37. Nedjimi B (2018) Heavy metal tolerance in two Algerian saltbushes: a review on plant responses to cadmium and role of calcium in its mitigation. In: Hasanuzzaman M, Fujita M, Oku H, Nahar K, Hawrylak-Nowak B (eds) Plant nutrients and abiotic stress tolerance. Springer, Singapore, pp 205–220CrossRefGoogle Scholar
  38. Neugschwandtner RW, Tlustoš P, Komárek M, Száková J (2008) Phytoextraction of Pb and cd from a contaminated agricultural soil using different EDTA application regimes: laboratory versus field scale measures of efficiency. Geoderma 144:446–454CrossRefGoogle Scholar
  39. Park JH, Choppala GK, Bolan NS, Chung JW, Chuasavathi T (2011) Biochar reduces the bioavailability and phytotoxicity of heavy metals. Plant Soil 348:439–451CrossRefGoogle Scholar
  40. Poovaiah BW (1993) Biochemical and molecular aspects of calcium action. Acta Hortic 326:139–145CrossRefGoogle Scholar
  41. Poovaiah BW, Reddy AS (1987) Calcium messenger system in plants. Crit Rev Plant Sci 6:47–103CrossRefGoogle Scholar
  42. Qin S, Sun X, Hu C, Tan Q, Zhao X, Xu S (2017) Effects of tungsten on uptake, transport and subcellular distribution of molybdenum in oilseed rape at two different molybdenum levels. Plant Sci 256:87–93CrossRefGoogle Scholar
  43. Rajkumar M, Sandhya S, Prasad MNV, Freitas H (2012) Perspectives of plant-associated microbes in heavy metal phytoremediation. Biotechnol Adv 30:1562–1574CrossRefGoogle Scholar
  44. Rygol J, Zimmermann U (1990) Radial and axial turgor pressure measurements in individual root cells of Mesembryanthemum crystallinum grown under various saline conditions. Plant Cell Environ 13:15–26CrossRefGoogle Scholar
  45. Shahid M, Ferrand E, Schreck E, Dumat C (2013) Behavior and impact of zirconium in the soil-plant system: plant update and phytotoxicity. Rev Environ Contam Toxicol 221:107–127Google Scholar
  46. Sun B, Zhao FJ, Lombi E, Mcgrath SP (2001) Leaching of heavy metals from contaminated soils using EDTA. Environ Pollut 113:111–120CrossRefGoogle Scholar
  47. Tessier A, Campbell PGC, Bisson M (1979) Sequential extraction procedure for the speciation of particulate trace metals. Anal Chem Anal Chem 51:844–851CrossRefGoogle Scholar
  48. Tester M (1990) Plant ion channels: whole-cell and single-channel studies. New Phytol 114:305–340CrossRefGoogle Scholar
  49. Udovic M, Lestan D (2009) Pb, Zn and cd mobility, availability and fractionation in aged soil remediated by EDTA leaching. Chemosphere 74:1367–1373CrossRefGoogle Scholar
  50. Vandenbosschea M, Jimeneza M, Casettaa M, Traisnela M (2015) Remediation of heavy metals by biomolecules: a review. Environ Sci Technol 45:1644–1704CrossRefGoogle Scholar
  51. Wang Y, Shen H, Xu L, Zhu XW, Li C, Zhang W, Xie Y, Gong YQ, Liu LW (2015) Transport, ultrastructural localization, and distribution of chemical forms of lead in radish (Raphanus sativus L.). Front Plant Sci 6:293–305Google Scholar
  52. Wang QY, Liu JS, Hu B (2016a) Integration of copper subcellular distribution and chemical forms to understand copper toxicity in apple trees. Environ Exp Bot 123:125–131CrossRefGoogle Scholar
  53. Wang SL, Nan ZR, Prete D, Ma JM, Liao Q, Zhang Q (2016b) Accumulation, transfer, and potential sources of mercury in the soil-wheat system under field conditions over the loess plateau Northwest China. Sci Total Environ 568:245–252CrossRefGoogle Scholar
  54. Wightwick AM, Mollah MR, Partington DL, Allinson G (2008) Copper fungicide residues in Australian vineyard soils. J Agric Food Chem 56:2457–2464CrossRefGoogle Scholar
  55. Wu FB, Dong J, Cai Y, Chen F, Zhang GP (2007) Differences in Mn uptake and subcellular distribution in different barley genotypes as a response to cd toxicity. Sci Total Environ 385:228–234CrossRefGoogle Scholar
  56. Xin JL, Huang BF (2013) Subcellular distribution and chemical forms of cadmium in two hot pepper cultivars differing in cadmium accumulation. J Agric Food Chem 62:508–515CrossRefGoogle Scholar
  57. Yang M, Xiao XY, Miao XF, Guo ZH, Wang FY (2012) Effect of amendments on growth and metal uptake of giant reed (Arundo donax L.) grown on soil contaminated by arsenic, cadmium and lead. Trans Nonferrous Metal Soc 22:1462–1469CrossRefGoogle Scholar
  58. Zhang P, Sun H, Yu L, Sun T (2013) Adsorption and catalytic hydrolysis of carbaryl and atrazine on pig manure-derived biochars: impact of structural properties of biochars. J Hazard Mater 244-245:217–224CrossRefGoogle Scholar
  59. Zhao YF, Wu JF, Shang DR, Ning JS, Zhai YX, Sheng XF, Ding HY (2015) Subcellular distribution and chemical forms of cadmium in the edible seaweed, Porphyra yezoensis. Food Chem 168:48–54CrossRefGoogle Scholar
  60. Zhou CF, Huang MY, Li Y, Luo JW, Cai LP (2016) Changes in subcellular distribution and antioxidant compounds involved in Pb accumulation and detoxification in Neyraudia reynaudiana. Environ Sci Pollut Res 23:21794–21804CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Na Liu
    • 1
    • 2
  • Jiulan Dai
    • 2
  • Haoqi Tian
    • 1
  • Huan He
    • 3
  • Yuen Zhu
    • 1
    Email author
  1. 1.College of Environmental and Resource SciencesShanxi UniversityTaiyuanChina
  2. 2.Environment Research InstituteShandong UniversityQingdaoChina
  3. 3.Department of Biology, Terrestrial Ecology SectionCopenhagen UniversityCopenhagenDenmark

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