Abstract
The present study aimed to investigate the response of chickpea (Cicer arietinum L.) seedlings to increasing concentrations (0, 10, 50, 100, 200, 500, 1000 µM) of cadmium (Cd) and to explore the potential protective effects of exogenous effectors against Cd-induced toxicity. Exposure of the germinating seeds to Cd significantly hindered the growth of the embryonic axis, resulting in dose-dependent reductions in elongation and dry biomass production. The Cd-induced growth delay might be attributable to the alteration of cotyledonary reserve mobilization. To assess the impact of exogenous effectors, namely magnesium (MgCl2, 10 mM), ascorbic acid (ASC, 1 µM), ethylene diamine tetraacetic acid (EDTA, 100 µM), and citric acid (100 µM), seeds were pretreated by imbibition in effector solutions for 12 h or were exposed to a combination of 200 µM Cd along with the effectors for 6 days. Pretreatment with ASC and EDTA effectively mitigated Cd-induced growth inhibition, leading to 28% and 36% increases in epicotyl fresh weight, respectively, without affecting the radicle and epicotyl lengths. The combined treatment with Cd and exogenous effectors, particularly ASC and EDTA, was more efficient at ameliorating seedling growth. Furthermore, all tested effectors countered the Cd-induced accumulation of H2O2 in chickpea tissues when applied concomitantly with the metal. The ameliorating effect of exogenous ASC and EDTA on the cellular redox state appears to be correlated with the enhancement of ascorbate peroxidase (APX) activity and the upregulation of the expression of its encoding gene. Conversely, catalase (CAT) activity was reduced by effector application, while the expression of its encoding gene was upregulated by ASC treatment. Overall, the present findings highlight the efficacy of ASC and EDTA as the most effective exogenous agents in protecting germinating chickpea seeds from Cd-imposed growth reduction and oxidative stress.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Explore related subjects
Discover the latest articles, news and stories from top researchers in related subjects.Data availability
The datasets generated and analyzed during the current study are available from the corresponding author on reasonable request.
References
Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126
Agami RA, Mohamed GF (2013) Exogenous treatment with indole-3-acetic acid and salicylic acid alleviates cadmium toxicity in wheat seedlings. Ecotoxicol Environ Saf 94:164–171. https://doi.org/10.1016/j.ecoenv.2013.04.013
Alexieva V, Sergiev I, Mapelli S, Karanov E (2001) The effect of drought and ultraviolet radiation on growth and stress markers in pea and wheat. Plant Cell Environ 24(12):1337–1344. https://doi.org/10.1046/j.1365-3040.2001.00778.x
Alshegaihi RM, Mfarrej MFB, Saleem MH et al (2023) Effective citric acid and EDTA treatments in cadmium stress tolerance in pepper (Capsicum annuum L.) seedlings by regulating specific gene expression. South Afr J Bot 159:367–380. https://doi.org/10.1016/j.sajb.2023.06.024
Anjum NA, Hasanuzzaman M, Hossain MA et al (2015) Jacks of metal/metalloid chelation trade in plants—an overview. Front Plant Sci 6:192. https://doi.org/10.3389/fpls.2015.00192
Anwar S, Shafiq F, Nisa Z et al (2021) Effect of cadmium stress on seed germination, plant growth and hydrolyzing enzymes activities in mungbean seedlings. J Seed Sci 43:e202143042. https://doi.org/10.1590/2317-1545v43256006
Ben Massoud M, Karmous I, Ferjani EE, Chaoui A (2018) Alleviation of copper toxicity in germinating pea seeds by IAA, GA3, Ca and citric acid. J Plant Interact 13:21–29. https://doi.org/10.1080/17429145.2017.1410733
Ben Massoud M, Sakouhi L, Chaoui A (2019) Effect of plant growth regulators, calcium and citric acid on copper toxicity in pea seedlings. J Plant Nutr 42:1230–1242. https://doi.org/10.1080/01904167.2019.1609506
Betti C, Della Rovere F, Piacentini D et al (2021) Jasmonates, ethylene and brassinosteroids control adventitious and lateral rooting as stress avoidance responses to heavy metals and metalloids. Biomolecules 11:77. https://doi.org/10.3390/biom11010077
Bhanuprakash K, Yogeesha HS (2016) Seed priming for abiotic stress tolerance: an overview. In: Rao NKS, Shivashankara KS, Laxman RH (eds) Abiotic stress physiology of horticultural crops. Springer India, New Delhi, pp 103–117. https://doi.org/10.1007/s11738-012-1186-5
Bhat JA, Basit F, Alyemeni MN et al (2023) Gibberellic acid mitigates nickel stress in soybean by cell wall fixation and regulating oxidative stress metabolism and glyoxalase system. Plant Physiol Biochem 198:107678. https://doi.org/10.1016/j.plaphy.2023.107678
Bolan N, Mahimairaja S, Kunhikrishnan A, Naidu R (2013) Sorption–bioavailability nexus of arsenic and cadmium in variable-charge soils. J Hazard Mater 261:725–732. https://doi.org/10.1016/j.jhazmat.2012.09.074
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 72:248–254. https://doi.org/10.1016/0003-2697(76)90527-3
Chen ZC, Yamaji N, Motoyama R et al (2012) Up-regulation of a magnesium transporter gene OsMGT1 is required for conferring aluminum tolerance in rice. Plant Physiol 159:1624–1633. https://doi.org/10.1104/pp.112.199778
Chen Q, Kan Q, Wang P et al (2015) Phosphorylation and interaction with the 14–3-3 protein of the plasma membrane H+-ATPase are involved in the regulation of magnesium-mediated increases in aluminum-induced citrate exudation in broad bean (Vicia faba. L). Plant Cell Physiol 56:1144–1153. https://doi.org/10.1093/pcp/pcv038
Chen HC, Zhang SL, Wu KJ et al (2020) The effects of exogenous organic acids on the growth, photosynthesis and cellular ultrastructure of Salix variegata Franch. under Cd stress. Ecotoxicol Environ Saf 187:109790. https://doi.org/10.1016/j.ecoenv.2019.109790
Chou TS, Chao YY, Huang WD et al (2011) Effect of magnesium deficiency on antioxidant status and cadmium toxicity in rice seedlings. J Plant Physiol 168:1021–1030. https://doi.org/10.1016/j.jplph.2010.12.004
Cui J, Liu T, Li Y, Li F (2018) Selenium reduces cadmium uptake into rice suspension cells by regulating the expression of lignin synthesis and cadmium-related genes. Sci Total Environ 644:602–610. https://doi.org/10.1016/j.scitotenv.2018.07.002
Ernst W (1998) Effects of heavy metals in plants at the cellular and organismic level ecotoxicology. In: Gerrit S, Bernd M III (eds) Bioaccumulation and biological effects of chemicals. Wiley and Spektrum Akademischer Verlag, Berlin, pp 587–620
Ghori N-H, Ghori T, Hayat MQ et al (2019) Heavy metal stress and responses in plants. Int J Environ Sci Technol 16:1807–1828. https://doi.org/10.1007/s13762-019-02215-8
Guo J, Lv X, Jia H et al (2020) Effects of EDTA and plant growth-promoting rhizobacteria on plant growth and heavy metal uptake of hyperaccumulator Sedum alfredii Hance. J Environ Sci 88:361–369. https://doi.org/10.1016/j.jes.2019.10.001
Hédiji H, Djebali W, Cabasson C et al (2010) Effects of long-term cadmium exposure on growth and metabolomic profile of tomato plants. Ecotoxicol Environ Saf 73:1965–1974. https://doi.org/10.1016/j.ecoenv.2010.08.014
Huda AKMN, Hossain M, Mukta RH et al (2021) EDTA-enhanced Cr detoxification and its potential toxicity in rice (Oryza sativa L.). Plant Stress. https://doi.org/10.1016/j.stress.2021.100014
Jacquemond M, Verdin E, Dalmon A et al (2009) Serological and molecular detection of Tomato chlorosis virus and Tomato infectious chlorosis virus in tomato. Plant Pathol 58:210–220. https://doi.org/10.1111/j.1365-3059.2008.01959.x
Karmous I, Khadija J, Chaoui A, Ferjani EE (2012) Proteolytic activities in Phaseolus vulgaris cotyledons under copper stress. Physiol Mol Biol Plants 18:337–343. https://doi.org/10.1007/s12298-012-0128-4
Kashem MDA, Kawai S (2007) Alleviation of cadmium phytotoxicity by magnesium in Japanese mustard spinach. Soil Sci Plant Nutr 53:246–251. https://doi.org/10.1111/j.1747-0765.2007.00129.x
Kaur R, Yadav P, Thukral AK et al (2018) Castasterone and citric acid supplementation alleviates cadmium toxicity by modifying antioxidants and organic acids in Brassica juncea. J Plant Growth Regul 37:286–299. https://doi.org/10.1007/s00344-017-9727-1
Kharbech O, Ben Massoud M, Sakouhi L et al (2020) Exogenous application of hydrogen sulfide reduces chromium toxicity in maize seedlings by suppressing NADPH oxidase activities and methylglyoxal accumulation. Plant Physiol Biochem 154:646–656. https://doi.org/10.1016/j.plaphy.2020.06.002
Kharbech O, Sakouhi L, Mahjoubi Y et al (2022) Nitric oxide donor, sodium nitroprusside modulates hydrogen sulfide metabolism and cysteine homeostasis to aid the alleviation of chromium toxicity in maize seedlings (Zea mays L.). J Hazard Mater 424:127302. https://doi.org/10.1016/j.jhazmat.2021.127302
Kubier A, Wilkin RT, Pichler T (2019) Cadmium in soils and groundwater: a review. Appl Geochem 108:1–16. https://doi.org/10.1016/j.apgeochem.2019.104388
Li D, Xiao S, Ma W-N et al (2020) Magnesium reduces cadmium accumulation by decreasing the nitrate reductase-mediated nitric oxide production in Panax notoginseng roots. J Plant Physiol 248:153131. https://doi.org/10.1016/j.jplph.2020.153131
Li L, Wang S, Wu S et al (2023) Morphological and physiological indicators and transcriptome analyses reveal the mechanism of selenium multilevel mitigation of cadmium damage in Brassica juncea. Plants 12:1583. https://doi.org/10.3390/plants12081583
Liu J, Qiu G, Liu C et al (2022) Salicylic acid, a multifaceted hormone, combats abiotic stresses in plants. Life 12:886. https://doi.org/10.3390/life12060886
Liu W, Yu T, Cao H et al (2023) Effects of EDTA and organic acids on physiological processes, gene expression levels, and cadmium accumulation in Solanum nigrum under cadmium stress. J Soil Sci Plant Nutr 23:3823–3833. https://doi.org/10.1007/s42729-023-01302-7
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
Lugon-Moulin N, Ryan L, Donini P, Rossi L (2006) Cadmium content of phosphate fertilizers used for tobacco production. Agron Sustain Dev 26:5. https://doi.org/10.1051/agro:2006010
Mabrouk B, Kâab SB, Rezgui M et al (2019) Salicylic acid alleviates arsenic and zinc toxicity in the process of reserve mobilization in germinating fenugreek (Trigonella foenum-graecum L.) seeds. S Afr J Bot 124:235–243. https://doi.org/10.1016/j.sajb.2019.05.020
Mahmud JA, Hasanuzzaman M, Nahar K et al (2018) Insights into citric acid-induced cadmium tolerance and phytoremediation in Brassica juncea L.: Coordinated functions of metal chelation, antioxidant defense and glyoxalase systems. Ecotoxicol Environ Saf 147:990–1001. https://doi.org/10.1016/j.ecoenv.2017.09.045
Mahmud JA, Hasanuzzaman M, Nahar K et al (2019) EDTA reduces cadmium toxicity in mustard (Brassica juncea L.) by enhancing metal chelation, antioxidant defense and glyoxalase systems. Acta Agrobotanica 72(2):1722. https://doi.org/10.5586/aa.1772
Matamoros MA, Becana M (2021) Molecular responses of legumes to abiotic stress: post-translational modifications of proteins and redox signaling. J Exp Bot 72:5876–5892. https://doi.org/10.1093/jxb/erab008
Mihoub A, Chaoui A, El Ferjani E (2005) Biochemical changes associated with cadmium and copper stress in germinating pea seeds (Pisum sativum L.). C R Biol 328:33–41. https://doi.org/10.1016/j.crvi.2004.10.003
Mohammadi S, Pourakbar L, Siavash Moghaddam S, Popović-Djordjević J (2021) The effect of EDTA and citric acid on biochemical processes and changes in phenolic compounds profile of okra (Abelmoschus esculentus L.) under mercury stress. Ecotoxicol Environ Saf. https://doi.org/10.1016/j.ecoenv.2020.111607
Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in Spinach chloroplasts. Plant Cell Physiol 22:867–880. https://doi.org/10.1093/oxfordjournals.pcp.a076232
Nazir F, Fariduddin Q, Khan TA (2020) Hydrogen peroxide as a signalling molecule in plants and its crosstalk with other plant growth regulators under heavy metal stress. Chemosphere 252:126486. https://doi.org/10.1016/j.chemosphere.2020.126486
Osmolovskaya N, Dung VV, Kuchaeva L (2018) The role of organic acids in heavy metal tolerance in plants. Biol Commun 63:9–16. https://doi.org/10.21638/spbu03.2018.103
Podazza G, Rosa M, González JA et al (2006) Cadmium induces changes in sucrose partitioning, invertase activities, and membrane functionality in roots of rangpur Lime (Citrus limonia L. Osbeck). Plant Biol 8:706–714. https://doi.org/10.1055/s-2006-924171
Rahoui S, Chaoui A, Ben C et al (2015) Effect of cadmium pollution on mobilization of embryo reserves in seedlings of six contrasted Medicago truncatula lines. Phytochemistry 111:98–106. https://doi.org/10.1016/j.phytochem.2014.12.002
Rather BA, Mir IR, Masood A et al (2020) Nitric oxide pre-treatment advances seed germination and alleviates copper-induced photosynthetic inhibition in Indian mustard. Plants 9:776. https://doi.org/10.3390/plants9060776
Rengel Z, Bose J, Chen Q et al (2015) Magnesium alleviates plant toxicity of aluminium and heavy metals. Crop Pasture Sci 66:1298–1307. https://doi.org/10.1071/CP15284
Roychoudhury A, Ghosh S, Paul S et al (2016) Pre-treatment of seeds with salicylic acid attenuates cadmium chloride-induced oxidative damages in the seedlings of mungbean (Vigna radiata L. Wilczek). Acta Physiol Plant 38:1–18. https://doi.org/10.1007/s11738-015-2027-0
Sakouhi L, El Ferjani E (2022) Effects of excess copper on sunflower seedling growth, mineral nutrition, and cellular redox state. Euro-Mediterr J Environ Integr 7:583–591. https://doi.org/10.1007/s41207-022-00335-1
Sakouhi L, Kharbech O, Ben Massoud M et al (2021) Calcium and ethylene glycol tetraacetic acid mitigate toxicity and alteration of gene expression associated with cadmium stress in chickpea (Cicer arietinum L.) shoots. Protoplasma 258:849–861. https://doi.org/10.1007/s00709-020-01605-x
Sakouhi L, Kharbech O, Ben Massoud M et al (2022a) Exogenous oxalic acid protects germinating chickpea seeds against cadmium injury. J Soil Sci Plant Nutr 22:647–659. https://doi.org/10.1007/s42729-021-00675-x
Sakouhi L, Kharbech O, Ben Massoud M et al (2022b) Oxalic acid mitigates cadmium toxicity in Cicer arietinum L. germinating seeds by maintaining the cellular redox homeostasis. J Plant Growth Regul 41:697–709. https://doi.org/10.1007/s00344-021-10334-1
Sakouhi L, Kadri O, Werghi S et al (2023a) Seed pretreatment with melatonin confers cadmium tolerance to chickpea seedlings through cellular redox homeostasis and antioxidant gene expression improvement. Environ Sci Pollut Res. https://doi.org/10.1007/s11356-023-27562-5
Sakouhi L, Mahjoubi Y, Labben A et al (2023b) Effects of cadmium–selenium interaction on glyoxalase and antioxidant systems of Pisum sativum germinating seeds. J Plant Growth Regul 42:3084–3099. https://doi.org/10.1007/s00344-022-10772-5
Sakouhi L, Werghi S, Ben Massoud M et al (2023c) Comparative study of different melatonin application effects on antioxidant system in cadmium-exposed chickpea seedlings. J Soil Sci Plant Nutr. https://doi.org/10.1007/s42729-023-01418-w
Saleem MH, Ali S, Rehman M et al (2020) Individual and combined application of EDTA and citric acid assisted phytoextraction of copper using jute (Corchorus capsularis L) seedlings. Environ Technol Innov 19:100895. https://doi.org/10.1016/j.eti.2020.100895
Sfaxi-Bousbih A, Chaoui A, El Ferjani E (2010) Cadmium impairs mineral and carbohydrate mobilization during the germination of bean seeds. Ecotoxicol Environ Saf 73:1123–1129. https://doi.org/10.1016/j.ecoenv.2010.01.005
Shen C, Fu HL, Liao Q et al (2021) Transcriptome analysis and physiological indicators reveal the role of sulfur in cadmium accumulation and transportation in water spinach (Ipomoea aquatica Forsk.). Ecotoxicol Environ Saf 225:112787. https://doi.org/10.1016/j.ecoenv.2021.112787
Široká B, Huttová J, Tamas L et al (2004) Effect of cadmium on hydrolytic enzymes in maize root and coleoptile. Biologia 59(4):513–517
Smirnoff N, Arnaud D (2019) Hydrogen peroxide metabolism and functions in plants. New Phytol 221:1197–1214. https://doi.org/10.1111/nph.15488
Tahjib-Ul-Arif M, Zahan MI, Karim MM et al (2021) Citric acid-mediated abiotic stress tolerance in plants. Int J Mol Sci 22:7235. https://doi.org/10.3390/ijms22137235
Tang M, Xu L, Wang Y et al (2021) Melatonin-induced DNA demethylation of metal transporters and antioxidant genes alleviates lead stress in radish plants. Horticult Res 8:124. https://doi.org/10.1038/s41438-021-00561-8
Ullah HA, Javed F, Wahid A, Sadia B (2016) Alleviating effect of exogenous application of ascorbic acid on growth and mineral nutrients in cadmium stressed barley (Hordeum vulgare) seedlings. Int J Agric Biol 18:73–79. https://doi.org/10.17957/IJAB/15.0064
Vaculík M, Konlechner C, Langer I et al (2012) Root anatomy and element distribution vary between two Salix caprea isolates with different Cd accumulation capacities. Environ Pollut 163:117–126. https://doi.org/10.1016/j.envpol.2011.12.031
Zhang F, Liu M, Li Y et al (2019) Effects of arbuscular mycorrhizal fungi, biochar and cadmium on the yield and element uptake of Medicago sativa. Sci Total Environ 655:1150–1158. https://doi.org/10.1016/j.scitotenv.2018.11.317
Zhou J, Cheng K, Huang G et al (2020) Effects of exogenous 3-indoleacetic acid and cadmium stress on the physiological and biochemical characteristics of Cinnamomum camphora. Ecotoxicol Environ Saf 191:109998. https://doi.org/10.1016/j.ecoenv.2019.109998
Zhou Z, Wei C, Liu H et al (2022) Exogenous ascorbic acid application alleviates cadmium toxicity in seedlings of two wheat (Triticum aestivum L.) varieties by reducing cadmium uptake and enhancing antioxidative capacity. Environ Sci Pollut Res 29:21739–21750. https://doi.org/10.1007/s11356-021-17371-z
Zlobin IE, Danilova ED, Murgan OK et al (2023) Structurally different exogenic brassinosteroids protect plants under polymetallic pollution via structure-specific changes in metabolism and balance of cell-protective components. Molecules 28:2077. https://doi.org/10.3390/molecules28052077
Zulfiqar U, Ayub A, Hussain S et al (2022) Cadmium toxicity in plants: recent progress on morpho-physiological effects and remediation strategies. J Soil Sci Plant Nutr 22:212–269. https://doi.org/10.1007/s42729-021-00645-3
Acknowledgements
The authors are thankful to Professor Yoshiyuki Murata for providing gene primers.
Funding
This work was financially supported by the Tunisian Ministry of High Education and Scientific Research (LR18ES38).
Author information
Authors and Affiliations
Contributions
LS performed all laboratory experiments, statistically analyzed the data, and wrote the manuscript; AC supervised the experiments and revised the manuscript. All the authors have read and approved the final manuscript.
Corresponding author
Ethics declarations
Conflict of interest
All authors declare that they have no conflict of interest.
Additional information
Responsible Editor: Angela Cunha.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Sakouhi, L., Chaoui, A. Modulation of antioxidant defense by exogenous compounds in cadmium-stressed chickpea seedlings. Euro-Mediterr J Environ Integr 9, 127–139 (2024). https://doi.org/10.1007/s41207-023-00440-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s41207-023-00440-9