Modulating effect of EDTA and SDS on growth, biochemical parameters and antioxidant defense system of Dahlia variabilis grown under cadmium and lead-induced stress


  • Yahya ALZAHRANI King Abdulaziz University, Faculty of Science, Department of Biological Sciences, Jeddah (SA)
  • Hesham F. ALHARBY King Abdulaziz University, Faculty of Science, Department of Biological Sciences, Jeddah (SA)
  • Khalid R. HAKEEM King Abdulaziz University, Faculty of Science, Department of Biological Sciences, Jeddah (SA)
  • Hameed ALSAMADANY King Abdulaziz University, Faculty of Science, Department of Biological Sciences, Jeddah (SA)



assisted phytoremediation; cadmium; phytoremediation enhancers; phytoremediation; ROS


The present study investigated the influence of inorganic amendments viz., SDS (sodium dodecyl sulphate) and ethylenediaminetetraacetic acid (EDTA) in enhancing metal tolerance in plants. Seedlings of an important ornamental plant, Dahlia variabilis Cav. were grown under cadmium (Cd) and lead (Pb) stress. 30-days old seedlings were transferred to pots containing sterilized sand and supplemented with Hoagland’s medium. After 15 days of transplanting, four treatments (0, 10, 25, and 100 mg kg-1) of Cd and four treatments of Pb (0, 100, 500 and 5000 mg kg-1) were used with or without application of 2.0 mM SDS and 2.5 mM EDTA, separately and in combination. Seedlings were further grown for 60 days in culture media. Results revealed that both Cd and Pb significantly reduced plant growth, pigment content, and relative water content. Antioxidant enzymes viz., superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX), and catalase (CAT) along with protein and total soluble sugar contents showed a declining trend with an increase in Cd and Pb concentrations applied. The Cd and Pb treatment enhanced the production rate of reactive oxygen species (ROS) as depicted by the increased malondialdehyde (MDA) and hydrogen peroxide (H2O2) production in leaf. Inorganic amendments viz., EDTA+SDS applied either alone or in combination significantly alleviated Cd and Pb-induced toxic effects. However, a combination of EDTA+SDS showed significant results than used separately. These results revealed that the application of inorganic amendments in combination can enhance the phytoextraction capacity of the species studied. However, the effects of various amendments vary with the nature of the inorganic compound. The study suggests that the application of EDTA and SDS could be a useful strategy for enhancing the phytoextraction capability of Dahlia variabilis to remove Cd and Pb from contaminated soils.


Aebi H (1984). Catalase in vitro. Methods in Enzymology 105:121-126.

Ahmad P, Ozturk M, Gucel S (2012). Oxidative damage and antioxidants induced by heavy metal stress in two cultivars of mustard (Brassica juncea L.) plants. Fresenius Environmental Bulletin 21(10):2953-2961.

Ashraf M, Ozturk M, Ahmad MSA (2010a). Plant adaptation and phytoremediation. Springer, New York, pp 481.

Ashraf M, Ozturk M, Ahmad MSA (2010b). Toxins and their phytoremediation. In: Ashraf M, Ozturk M, Ahmad MSA (Eds.). Plant adaptation and phytoremediation. Springer, Dordrecht, pp 1-34.

Ashraf MY, Roohi M, Iqbal Z, Ashraf M, Ozturk M, Gucel S (2015). Cadmium (Cd) and lead (Pb) induced inhibition in growth and alteration in some biochemical attributes and mineral accumulation in mung bean [Vigna radiata (L.) Wilczek]. Communications in Soil Science and Plant Analysis 47(4):405-413.

Anjum NA, Adam V, Kizek R, Duarte AC, Pereira E, Iqbal M, … Ahmad I (2015). Nanoscale copper in the soil-plant system: Toxicity and underlying potential mechanisms. Environmental Research 138:306-325.

Anjum NA, Umar S, Iqbal M (2014). Assessment of cadmium accumulation, toxicity and tolerance in Brassicaceae and Fabaceae plants - implications for phytoremediation. Environmental Science and Pollution Research 21(17):10286-10293.

Ansari MKA, Anjum NA, Ahmad A, Umar S, Iqbal M (2012). Heavy metals in soil and plants: An overview of arsenic, cadmium, chromium and mercury. In: Anjum NA, Umar S, Ahmad A (Eds.) Oxidative stress in plants: causes, consequences and tolerance. I.K. International Publishing House, New Delhi, India, pp 499-518.

Aziz MA, Ahmad HR, Corwin DL, Sabir M, Ozturk M, Hakeem KR (2016). Influence of farmyard manure on retention and availability of nickel, zinc and lead in metal-contaminated calcareous loam soils. Journal of Environmental Engineering and Landscape Management 25(3):289-296.

Bagheri R, Bashir H, Ahmad J, Iqbal M, Qureshi MI (2015). Spinach (Spinacia oleracea L.) modulates its proteome differentially in response to salinity, cadmium and their combination stress. Plant Physiology and Biochemistry 97: 235-245.

Bareen FE (2012). Chelate assisted phytoextraction using oil seed brassicas. Environmental Pollution 21:289-311.

Bashir H, Qureshi MI, Ibrahim AM, Iqbal M (2015) Chloroplast and photosystems: impact of cadmium and iron deficiency. Photosynthetica 53 (3): 321-335.

Bates LS, Walderen RD, Taere ID (1973). Rapid determination of free proline for water-stress studies. Plant Soil 39:205-207.

Bohnert HJ, Nelson DE, Jensen RG (1995). Adaptations to environmental stresses. Plant Cell 7:1099-1111.

Bradford MM (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dyes binding. Analytical Biochemistry 72:248-254.

Brunet J, Repellin A, Varrault G, Terrync N, Zuily-Fodil Y (2008). Lead accumulation in the roots of grass pea (Lathyrussativus): A novel plant for phytoremediation systems? Comptes Rendus Biologies 331:859-864.

Chance M, Maehly AC (1955). Assay of catalases and peroxidases. Methods in Enzymology 2:764-817.

Chen J, Shafi M, Li S, Wang Y, Wu J, Ye Z, … Liu D (2015). Copper induced oxidative stresses, antioxidant responses and phytoremediation potential of Moso bamboo (Phyllostachys pubescens). Scientific Reports 5:13554.

Chen J, Shiyab S, Han FX, Monts DL, Waggoner AW, Su ZY (2009). Bioaccumulation and physiological effects of mercury in Pteris vittata and Nephrolepi sexaltata. Ecotoxicology 18:110-121.

Chigbo C, Batty L (2013). Effect of EDTA and citric acid on phytoremediation of Cr-B [a] P-co contaminated soil. Environmental Science and Pollution Research 20(12):8955-8963.

Dixit P, Mukherjee PK, Ramachandran V, Eapen S (2011). Glutathione transferase from Trichoderma virens enhances cadmium tolerance without enhancing its accumulation in transgenic Nicotiana tabacum. PLoS ONE 6:e16360.

Dong J, Wu FB, Zhang GP (2006). Influence of cadmium on antioxidant capacity and four microelement concentrations in tomato seedlings (Lycopersicon esculentum). Chemosphere 64:1659-1666.

Duruibe JO, Ogwuegbu MOC, Egwurugwu JN (2007). Heavy metal pollution and human biotoxic effects. International Journal of Physical Sciences 2(5):112-118.

Ehsan S, Ali S, Noureen S, Mehmood K, Farid M, Ishaque W, … Rizwan M (2014) Citric acid assisted phytoremediation of Cd by Brassica napus L. Ecotoxicology and Environmental Safety 106:164-172.

Evangelou VP, Marsi M (2001). Composition and metal ion complexation behavior of humic fractions derived from corn tissue. Plant Soil 229:13-24.

Farid M, Ali S, Shakoor MB, Bharwana SA, Rizvi H, Ehsan S, … Hannan F (2013). EDTA assisted phytoremediation of cadmium, lead and zinc. International Journal of Agronomy and Plant Production 4(11):2833-2846.

Feng-Tao LI, Jian-Min QI, Gao-Yang Z, Li-Hui L, Ping-Ping F, Fen TA, Jian-Tang XU (2013). Effect of cadmium stress on the growth antioxidative enzymes and lipid peroxidation in two kenaf (Hibiscus cannabinus L.) plant seedlings. Journal of Integrative Agriculture 12:610-620.

Gajewska E, Skłodowska M (2010). Differential effect of equal copper, cadmium and nickel concentration on biochemical reactions in wheat seedlings. Ecotoxicology and Environmental Safety 73(5):996-1003.

Gill SS, Tuteja N (2010). Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiology and Biochemistry 48:909-930.

Gallego SM, Pena LB, Barcia RA, Azpilicueta CE, Iannone MF, Rosales EP, … Benavides MP (2012). Unravelling cadmium toxicity and tolerance in plants: insight into regulatory mechanisms. Environmental and Experimental Botany 83:33-46.

Ghori NH, Ghori T, Hayat MQ, Imadi SR, Gul A, Altay V, Ozturk M (2019). Heavy metal stress and responses in plants. International Journal of Environmental Science and Technology 16(3):1807-1828.

Habiba U, Ali S, Farid M, Sharkoor MB, Rizwan M, Ibrahim M, … Ali B (2015). EDTA enhanced plant growth, antioxidant defense system, and phytoextraction of copper by Brassica napus L. Environmental Science and Pollution Research 22(2):1534-1544.

Hajiboland R, Rad SB, Barceló J, Poschenrieder C (2013). Mechanisms of aluminum-induced growth stimulation in tea (Camellia sinensis). Journal of Plant Nutrition and Soil Science 176:616-625.

Hakeem KR, Alharby HF, Rehman R (2019). Antioxidative defense mechanism against lead-induced phytotoxicity in Fagopyrum kashmirianum. Chemosphere 216:595-604.

Hakeem KR, Sabir M, Ozturk M, Mermut A (2015). Soil remediation and plants: prospects and challenges. Elsevier, London, pp 724.

Hameed A, Qadri TN, Zaffar M, Siddiqi TO, Ozturk M, Altay V, Ahmad P (2017). Biochemical and nutritional responses of Abelmoschus esculentus L. exposed to mercury contamination. Feb-Fresenius Environmental Bulletin 26(10):5814-5823.

Han F, Shan XQ, Zhang J, Xie YN, Pei ZJ, Zhang SZ, … Wen B (2005). Organic acids promote the uptake of lanthanum by barley roots. New Phytologist 165:481-492.

Hasan, MM, Uddin MN, Ara-Sharmeen IF, Alharby H, Alzahrani Y, Hakeem KR, Zhang L (2019). Assisting phytoremediation of heavy metals using chemical amendments. Plants 8(9):295.

Hayat S, Hayat Q, Alyemeni MN, Wani AS, Pichtel J, Ahmad A (2012). Role of proline under changing environments: a review. Plant Signaling & Behavior 7(11):1456-1466.

Heath RL, Packer L (1968). Photo-peroxidation in isolated chloroplasts: I. Kinetics and stoichiometry of fatty acid peroxidation. Archives of Biochemistry and Biophysics 125:189-198.

Hoagland DR, Arnon DI (1950). The water-culture for growing plants without soil. Circular. California Agricultural Experiment Station 347(2):32.

Hu R, Sunc K, Suc X, Pana Y, Zhanga Y, Wanga X (2012). Physiological responses and tolerance mechanisms to Pb in two xerophils: Salsolapasserina bunge and Chenopodium album L. Journal of Hazardous Materials 205(206):131-138.

Iqbal M, Ahmad A, Ansari MKA, Qureshi MI, Aref MI, Khan PR, … Hakeem KR (2015). Improving the phytoextraction capacity of plants to scavenge metal(loid)-contaminated sites. Soil & Environment 23(1): 44-65.

Jabeen R, Ahmad A, Iqbal M (2009). Phytoremediation of heavy metals: Physiological and molecular aspects. The Botanical Review 75:339-364.

Jaishankar M, Tseten T, Anbalagan N, Mathew BB, Beeregowda KN (2014). Toxicity, mechanism and health effects of some heavy metals. Interdisciplinary Toxicology 7(2):60-72.

Kambhampati MS (2013). EDTA enhanced phytoremediation of copper contaminated soils using chickpea (Cicera eritinum L.). Bulletin of Environmental Contamination and Toxicology 91(3):310-313.

Khudsar T, Iqbal M (2001). Cadmium-induced change in leaf epidermes, photosynthetic rate and pigment concentrations in Cajanus cajan. Biologia Plantarum 44:59-64.

Kupper H, Kupper F, Spiller M (1998). In situ detection of heavy metal substituted chlorophylls in water plants. Photosynthesis Research 58:123-133.

Lichtenthaler HK, Wellburn AR (1983). Determination of total carotenoids and chlorophylls a and b of leaf extracts in different solvents. Biochemical Society Transaction 11:591-603.

Liu D, Wang X, Chen Z, Xu H, Wang Y (2010). Influence of mercury on chlorophyll content in winter wheat and mercury bioaccumulation. Plant, Soil and Environment 56:139-143.

Ma MF, Zheng SJ, Hiradate S, Matsumoto H (1997). Detoxifying aluminum with buckwheat. Nature 390:569-570.

Malar S, Sahi SV, Favas PJC, Venkatachalam P (2015). Mercury heavy-metal-induced physiochemical changes and genotoxic alterations in water hyacinths [Eichhornia crassipes (Mart.)]. Environmental Science and Pollution Research 22:4597-4608.

Mani D, Kumar C, Patel NK (2014). Hyperaccumulator oilcake manure as an alternative for chelate-induced phytoremediation of heavy metals contaminated alluvial soils. International Journal of Phytoremediation 17(1-6):256-63.

Marschner H (1995). Mineral nutrition in higher plants. 2nd edn. Academic, London.

Marchiol L, Assolari S, Sacco P, Zerbi G (2004). Phytoextraction of heavy metals by canola (Brassica napus) and radish (Raphanus sativus). Environmental Pollution 132(1):21-27.

Nouri J, Lorestani B, Yousefi N, Khorasani N, Hasani AH, Seif S, Cheraghi M (2011). Phytoremediation potential of native plants grown in the vicinity of Ahangaran lead-zinc mine Hamedan, Iran. Environmental Earth Sciences 62(3):639-644.

Nor M, Cheng H (1986). Chemical speciation and bioavailability of Cu: uptake and accumulation by Eichornia. Environmental Toxicology and Chemistry: An International Journal 5:941-947.

Ozturk M (1989). Plants and pollutants in developed and developing countries. Ege Univ. Press, Izmir, pp 759.

Ozturk M, Ashraf M, Aksoy A, Ahmad MSA (2015a). Phytoremediation for green energy. Springer, New York.

Ozturk M, Ashraf M, Aksoy A, Hakeem KR, Ahmad MSA (2015b). Plants, pollutants and remediation. Springer, New York.

Ozturk M, Altay V, Karahan F (2017). Studies on trace elements in Glycyrrhiza taxa distributed in Hatay-Turkey. International Journal of Plant and Environment 3(2):01-07.

Pirzadah TB, Malik B, Tahir I, Kumar M, Varma A, Rehman RU (2014). Phytoremediation: an eco-friendly green technology for pollution prevention, control and remediation. In: Hakeem KR, Sabir M, Ozturk M, Mermut AH (Eds.). Soil remediation and plants: prospects and challenges. Elsevier publications, USA, pp 107-122.

Placek A, Grobelak A, Kacprzak M (2016). Improving the phytoremediation of heavy metals contaminated soil by use of sewage sludge. International Journal of Phytoremediation 18(6):605-618.

Plata JS, Villasante CO, Flores-C´aceres ML, Escobar C, del Campo FF, Hernandez LE (2009). Differential alterations of antioxidant defenses as bio-indicators of mercury and cadmium toxicity in Alfalfa. Chemosphere 77(7):946-954.

Prasad DDK, Prasad ARK (1987). Altered o-aminolevulinic acid metabolism by lead and mercury in germination seedlings of bajra (Pennisetum typoideum). Journal of Plant Physiology 127:241-249.

Quartacci M, Pinzino C, Sgherri C, Dalla VF, Navari-Izz F (2000). Growth in excess copper induces changes in the lipid composition and fluidity of PS-II enriched membrane in wheat. Physiologia Plantarum 108(1):87-93.

Qureshi MI, Israr M, Abdin MZ, Iqbal M (2005). Responses of Artemisia annua L. to lead and salt-induced oxidative stress. Environmental and Experimental Botany 53:185-193.

Qureshi MI, Abdin MZ, Qadir S, Iqbal M (2007). Lead-induced oxidative stress and metabolic alterations in Cassia angustifolia Vahl. Biologia Plantarum 51: 121-128.

Rastgoo L, Alemzadeh A (2011). Biochemical responses of Gouan (Aeluropus littoralis) to heavy metals stress. Australian Journal of Crop Science 5(4):375-383.

Rizwan M, Ali S, Adrees M, Rizvi H, Rehman MZ, Hannan F, … Ok YS (2016). Cadmium stress in rice: toxic effects, tolerance mechanisms and management: A critical review. Environmental Science and Pollution Research 23(18):17859-17879.

Rosa M, Prado C, Podazza G, Interdonato R, González JA, Hilal M, Prado FE (2009). Soluble sugars-metabolism, sensing and abiotic stress: A complex network in the life of plants. Plant Signaling & Behavior 4(5):388-393.

Sabir M, Waraich EA, Hakeem KR, Ozturk M, Ahmad HR, Shahid M (2015b). Phytoremediation: mechanisms and adaptations. In: Hakeem K, Sabir M, Ozturk M, Mermut A (eds). Soil remediation and plants: prospects and challenges. Elsevier, New York, pp 85-105.

Sahu, GK, Upadhyay S, Sahoo BB (2012) Mercury induced phytotoxicity and oxidative stress in wheat (Triticum aestivum L.) plants. Physiology and Molecular Biology of Plants 18(1):21-31.

Saifullah Khan MN, Iqbal M, Naeem A, Bibi S, Waraich EA, Dahlawi S (2016). Elemental sulfur improves growth and phytoremediative ability of wheat grown in lead-contaminated calcareous soil. International Journal of Phytoremediation 18:1022-1028.

Sarwar N, Imran M, Shaheen MR, Ishaque W, Kamran MA, Matloob A, … Hussaine S (2017). Phytoremediation strategies for soils contaminated with heavy metals: Modifications and future perspectives. Chemosphere 710-721.

Saxena G, Purchase D, Mulla SI, Saratale GD, Bharagava RN (2019). Phytoremediation of heavy metal-contaminated sites: eco-environmental concerns, field studies, sustainability issues, and future prospects. In: de Voogt P (Eds.). Rev EnvContTox Vol 249. Reviews of Environmental Contamination and Toxicology (Continuation of Residue Reviews), vol 249. Springer, Cham.

Schützendübel A, Polle A (2002). Plant responses to abiotic stresses: heavy metal-induced oxidative stress and protection by mycorrhization. Journal of Experimental Botany 53:1351-1365.

Shu, X, Yin LY, Zhang QF, Wang WB (2015). Effect of Pb toxicity on leaf growth, antioxidant enzyme activities, and photosynthesis in cuttings and seedlings of Jatropha curcas L. Environmental Science and Pollution Research 19(3):893-902.

Singh J, Kalamdhad AS (2011). Effects of heavy metals on soil, plants, human health and aquatic life. International Journal of Research in Chemistry and Environment 1(2):15-21.

Souza LA, Piotto FA, Nogueirol RC, Azevedo RA (2013). Use of non-hyperaccumulator plant species for the phytoextraction of heavy metals using chelating agents. Scientia Agricola 70(4):290-295.

Szczygłowskan M, Piekarska A, Konieczka P, Namiesnik J (2011). Use of brassica plants in the phytoremediation and biofumigation processes. International Journal of Molecular Sciences 12:7760-7771.

Székely A, Balota DA, Duchek JM, Nemoda Z, Vereczkei A, SasvariSzekely M (2011). Genetic factors of reaction time performance: DRD4 7 repeat allele. Genes Brain and Behavior 10(2):129-136.

Wang Y, Li Y, Ma C, Qiu D (2016). Gas exchange, photosystem II photochemistry, and the antioxidant system of longan plant (Dimocarpus longan Lour.) leaves in response to lead (Pb) stress. Plant Omics 9(4):240-247.

Wilkins DA (1957). A technique for the measurement of Pb tolerance in plants. Nature 180:37-38.

Wirosoedarmo R, Anugroho F, Hanggara SD, Gustinasari K (2018). Effect of adding chelating agents on the absorption of zinc from polluted soil sludge textile industrial waste by sunflower plant (Helianthus annuus L.). Applied and Environmental Soil Science.

Yu CW, Murphy TH, Lin CH (2003). Hydrogen peroxide-induces chilling tolerance in mung beans mediated through ABA independent glutathione accumulation. Functional Plant Biology 30:955-963.

Zhang H, Li YH, Hu LY,Wang SH, Zhang FQ, Hu KD (2008). Effects of exogenous nitric oxide donor on antioxidant metabolism in wheat leaves under aluminum stress. Russian Journal of Plant Physiology 55:469-474.

Zhang Y, Xu S, Yang S, ChenY (2015). Salicylic acid alleviates cadmium-induced inhibition of growth and photosynthesis through upregulating antioxidant defense system in two melon cultivars (Cucumismelo L.). Protoplasma 252:911-924.




How to Cite

ALZAHRANI, Y. ., ALHARBY, H. F. ., HAKEEM, K. R., & ALSAMADANY, H. . (2020). Modulating effect of EDTA and SDS on growth, biochemical parameters and antioxidant defense system of Dahlia variabilis grown under cadmium and lead-induced stress. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 48(2), 906–923.



Research Articles
DOI: 10.15835/nbha48211909

Most read articles by the same author(s)