Skip to main content
SpringerLink
Log in

Synergistic effects of exogenous glutathione and calcium on ascorbate–glutathione cycle and glutathione-associated enzymes upregulation under lead stress in Brassica napus L

  • Research Article
  • Published:
Environmental Science and Pollution Research Aims and scope Submit manuscript

Abstract

Heavy metals (HMs) such as lead (Pb) pose a significant threat to global food security due to their adverse effect on the health of crop plants. Calcium (Ca) and Glutathione (GSH) are signaling molecules to scavenge free radicals in HM-stressed plants. In this study, GSH and Ca’s role is examined in supporting canola seedlings against Pb toxicity. In a pot experiment, the administration of Glutathione (GSH, 0 and 100 µM) and/or calcium (CaCl2, 0 and 500 µM) in canola seedlings was examined under lead stress (0 and 100 µM of Pb(NO3)2. Compared with the control samples, Pb treatment increased MDA and H2O2 values by 61 and 53%, respectively, indicative of oxidative burst. However, using a combination of GSH and Ca lowered oxidative stress in Pb-stressed plants by an approximately twofold reduction in MDA and H2O2 content. Total PC content increased by 78% in Pb-stressed plants, suggesting that these chelating peptides diminish the damaging effects of Pb. Interestingly, further boosts in total PC levels were recorded in Pb-stressed plants treated with GSH and Ca concurrently. The addition of exogenous GSH and Ca to Pb-stressed canola plants limited Pb uptake and translocation and improved ascorbate–glutathione cycle performance. Moreover, compared to their separate usage, the co-treatment of exogenous GSH and Ca strengthened the GSH pool by increasing the activities of enzymes involved in GSH metabolism. The findings demonstrate that exogenous GSH and Ca modulate GSH synthesis, metabolism, and redox homeostasis synergistically to enhance resistance to oxidative stress generated by Pb.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

Data availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

References

  • Ahmad P, Sarwat M, Bhat NA, Wani MR, Kazi AG, Tran LSP (2015) Alleviation of cadmium toxicity in Brassica juncea L. (Czern. & Coss.) by calcium application involves various physiological and biochemical strategies. PloS one 10(1):114571

    Article  Google Scholar 

  • Ahmad P, Abd-Allah EF, Alyemeni MN, Wijaya L, Alam P, Bhardwaj R, Siddique KH (2018) Exogenous application of calcium to 24-epibrassinosteroid pre-treated tomato seedlings mitigates NaCl toxicity by modifying ascorbate–glutathione cycle and secondary metabolites. Sci. Rep 8(1):1–15

    Article  Google Scholar 

  • Ahmed N, Ehsan A, Danish S, Ali MA, Fahad S, Dawar K, Taban S, Akça H, Shah AA, Ansari MJ, Babur E (2022) Mitigation of lead (Pb) toxicity in rice cultivated with either ground water or wastewater by application of acidified carbon. J Environ Manag 307:114521

    Article  CAS  Google Scholar 

  • Ali B, Xu X, Gill RA, Yang S, Ali S, Tahir M, Zhou W (2014) Promotive role of 5-aminolevulinic acid on mineral nutrients and antioxidative defense system under lead toxicity in Brassica napus. Ind Crops Prod 52:617–626

    Article  CAS  Google Scholar 

  • Amooaghaie R, Enteshari S (2017) Role of two-sided crosstalk between NO and H2S on improvement of mineral homeostasis and antioxidative defense in Sesamum indicum under lead stress. Ecotoxicol Environ Saf 139:210–218

    Article  CAS  Google Scholar 

  • Aqeel U, Parwez R, Aftab T, Khan MMA, Naeem M (2022) Exogenous calcium repairs damage caused by nickel toxicity in fenugreek (Trigonella foenum-graecum L.) by strengthening its antioxidant defense system and other functional attributes. S Afr J Bot 150:153–160

    Article  CAS  Google Scholar 

  • Arianmehr M, Karimi N, Souri Z (2022) Exogenous supplementation of Sulfur (S) and Reduced Glutathione (GSH) Alleviates Arsenic Toxicity in Shoots of Isatis cappadocica Desv and Erysimum allionii L. Environ Sci Pollut Res 1–10. https://doi.org/10.1007/s11356-022-19477-4

  • Arnon DI (1949) Copper enzymes in isolated chloroplasts, polyphenol oxidase in Beta vulgaris L. Plant Physiol 24:1–15

    Article  CAS  Google Scholar 

  • Arnon DI, Hogland DR (1940) Crop production in artificial solutions and in soils with special references to factors influencing yields and absorption of inorganic nutrients. Soil Sci 50:463–484

    CAS  Google Scholar 

  • Aslam S, Gul N, Mir MA, Asgher M, Al-Sulami N, Abulfaraj AA, Qari S (2021) Role of jasmonates, calcium, and glutathione in plants to combat abiotic stresses through precise signaling cascade. Front Plant Sci 12:1172

    Article  Google Scholar 

  • Bashir S, Jan N, Wani UM, Raja V, John R (2022) Co-over expression of Ascorbate Glutathione pathway enzymes improve mercury tolerance in tomato. Plant Physiol Biochem 186:170–181

    Article  CAS  Google Scholar 

  • Bhargava P, Srivastava AK, Urmil S, Rai LC (2005) Phytochelatin plays a role in UV-B tolerance in N2-fixing cyanobacterium Anabaena doliolum. J Plant Physiol 162:1220–1225

    Article  CAS  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantification 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

  • Cakmak I, Marschner H (1992) Magnesium Deficiency and High Light Intensity Enhance Activities of Superoxide Dismutase, Ascorbate Peroxidise and Glutathione Reductase in Bean Leaves. Plant Physiolo 98:1222–1227

    Article  CAS  Google Scholar 

  • Chen F, Aqeel M, Maqsood MF, Khalid N, Irshad MK, Ibrahim M, Akhter N, Afzaal M, Ma J, Hashem M, Alamri S (2022) Mitigation of lead toxicity in Vigna radiata genotypes by silver nanoparticles. Environ Pollut 308:119606

    Article  CAS  Google Scholar 

  • Cobbett CS, May MJ, Howden R, Rolls B (1998) The glutathione deficient, cadmium-sensitive mutant, cad2-1, of Arabidopsis thaliana is deficient in γ-glutamylcysteine synthetase. Plant J 16:73–78

    Article  CAS  Google Scholar 

  • Del Longo OT, Gonzalez CA, Pastori GM, Tripps VS (1993) Antioxidant defenses under hyperoxygenic and hyperosmotic conditions in leaves of two lines of maize with differential sensitivity to drought. Plant Cell Physiol 34:1023–1028

    Google Scholar 

  • Ding S, Ma C, Shi W, Liu W, Lu Y, Liu Q, Luo ZB (2017) Exogenous glutathione enhances cadmium accumulation and alleviates its toxicity in Populus×canescens. Tree Physiol 37(12):1697–1712

    Article  CAS  Google Scholar 

  • El-Beltagi HS, Mohamed HI, Sofy MR (2020) Role of ascorbic acid, glutathione and proline applied as singly or in sequence combination in improving chickpea plant through physiological change and antioxidant defense under different levels of irrigation intervals. Molecules 25(7):1702

    Article  CAS  Google Scholar 

  • Elia AC, Galarini R, Taticchi MI, Dorr AJM, Mantilacci L (2003) Antioxidant responses and bioaccumulation in Ictalurus melas under mercury exposure. Ecotoxicol Environ Saf 55:162–167

    Article  CAS  Google Scholar 

  • Farooq MA, Hong Z, Islam F, Noor Y, Hannan F, Zhang Y, Ayyaz A, Mwamba TM, Zhou W, Song W (2021) Comprehensive proteomic analysis of arsenic induced toxicity reveals the mechanism of multilevel coordination of efficient defense and energy metabolism in two Brassica napus cultivars. Ecotoxicol Environ Saf 208:111744

    Article  CAS  Google Scholar 

  • Feng N, Yu M, Li Y, Jin D, Zheng D (2021) Prohexadione-calcium alleviates saline-alkali stress in soybean seedlings by improving the photosynthesis and up-regulating antioxidant defense. Ecotoxicol Environ Saf 220:112369

    Article  CAS  Google Scholar 

  • Griffith OW (1980) Determination of glutathione and glutathione disulphide using glutathione reductase and 2-vinyl pyridine. Anal Biochem 106:207–212

    Article  CAS  Google Scholar 

  • Guo J, Zhou R, Ren X, Jia H, Hua L, Xu H, Lv X, Zhao J, Wei T (2018) Effects of salicylic acid, Epi-brassinolide and calcium on stress alleviation and Cd accumulation in tomato plants. Ecotoxicol Environ Saf 157:491–496

    Article  CAS  Google Scholar 

  • Hakeem KR, Alharby HF, Pirzadah TB (2022) Exogenously applied calcium regulates antioxidative system and reduces cadmium-uptake in Fagopyrum esculentum. Plant Physiol Biochem 180:17–26

    Article  CAS  Google Scholar 

  • Hasanuzzaman M, Nahar K, Rahman A, Mahmud JA, Alharby HF, Fujita M (2018) Exogenous glutathione attenuates lead-induced oxidative stress in wheat by improving antioxidant defense and physiological mechanisms. J Plant Interact 13(1):203–212

    Article  CAS  Google Scholar 

  • Hossain MA, Asada K (1984) Purification of dehydroascorbate reductase from spinach and its characterization as thiol enzyme. Plant Cell Physiol 25:85–92

    CAS  Google Scholar 

  • Jahan A, Iqbal M, Shafiq F, Malik A, Javed MT (2021) Influence of foliar glutathione and putrescine on metabolism and mineral status of genetically diverse rapeseed cultivars under hexavalent chromium stress. Environ Sci Pollut Res 28(33):45353–45363

    Article  CAS  Google Scholar 

  • Jiang M, Zhang J (2002) Water stress-induced abscisic acid accumulation triggers the increased generation of reactive oxygen species and up-regulates the activities of antioxidant enzymes in maize leaves. Exp Bot 53:2401–2410

    Article  CAS  Google Scholar 

  • Kanu AS, Ashraf U, Mo Z, Sabir SUR, Baggie I, Charley CS, Tang X (2019) Calcium amendment improved the performance of fragrant rice and reduced metal uptake under cadmium toxicity. Environ Sci Pollut Res 26(24):24748–24757

    Article  CAS  Google Scholar 

  • Kaya C, Akram NA, Sürücü A, Ashraf M (2019) Alleviating effect of nitric oxide on oxidative stress and antioxidant defence system in pepper (Capsicum annuum L.) plants exposed to cadmium and lead toxicity applied separately or in combination. Sci Hortic 255:52–60

    Article  CAS  Google Scholar 

  • Kaya C, Ugurlar F, Farooq S, Ashraf M, Alyemeni MN, Ahmad P (2022) Combined application of asparagine and thiourea improves tolerance to lead stress in wheat by modulating AsA-GSH cycle, lead detoxification and nitrogen metabolism. Plant Physiol Biochem 190:119–132

    Article  CAS  Google Scholar 

  • Khan A, Khan S, Alam M, Khan MA, Aamir M, Qamar Z, Rehman ZU, Perveen S (2016) Toxic metal interactions affect the bioaccumulation and dietary intake of macro-and micro-nutrients. Chemosphere 146:121–128

    Article  CAS  Google Scholar 

  • Kohli SK, Handa N, Sharma A, Gautam V, Arora S, Bhardwaj R, Wijaya L, Alyemeni MN, Ahmad P (2018) Interaction of 24-epibrassinolide and salicylic acid regulates pigment contents, antioxidative defense responses, and gene expression in Brassica juncea L. seedlings under Pb stress. Environ Sci Pollut Res 25(15):15159–15173

    Article  CAS  Google Scholar 

  • Kumar A, Prasad MNV (2018) Plant-lead interactions: transport, toxicity, tolerance, and detoxification mechanisms. Ecotoxicol Environ Saf 166:401–418

    Article  CAS  Google Scholar 

  • Li GZ, Chen SJ, Li NY, Wang YY, Kang GZ (2021a) Exogenous glutathione alleviates cadmium toxicity in wheat by influencing the absorption and translocation of cadmium. Bull Environ Contam Toxicol 107(2):320–326

    Article  CAS  Google Scholar 

  • Li Z, Mei X, Li T, Yang S, Qin L, Li B, Zu Y (2021b) Effects of calcium application on activities of membrane transporters in Panax notoginseng under cadmium stress. Chemosphere 262:127905

    Article  CAS  Google Scholar 

  • Liu J, Zhang J, Kim SH, Lee HS, Marinoia E, Song WY (2021) Characterization of Brassica rapa metallothionein and phytochelatin synthase genes potentially involved in heavy metal detoxification. PLoS One 16(6):0252899

    Article  Google Scholar 

  • Ma L, Kong L, Gui R, Yang X, Zhang J, Gong Q, Qin D, Zhuang M, Ashraf U, Mo Z (2021) Application of hydrogen-rich water modulates physio-biochemical functions and early growth of fragrant rice under Cd and Pb stress. Environ Sci Pollut Res 28:58558–58569

    Article  CAS  Google Scholar 

  • Menhas S, Yang X, Hayat K, Ali A, Ali EF, Shahid M, Shaheen SM, Rinklebe J, Hayat S, Zhou P (2022) Melatonin enhanced oilseed rape growth and mitigated Cd stress risk: A novel trial for reducing Cd accumulation by bioenergy crops. Environ Pollut 308:119642

    Article  CAS  Google Scholar 

  • Mishra RK, Mishra G, Singh R, Parihar P, Kumar J, Srivastava PK, Prasad SM (2022) Managing arsenic (V) toxicity by phosphate supplementation in rice seedlings: modulations in AsA-GSH cycle and other antioxidant enzymes. Environ Sci Pollut Res 29(10):14418–14429

    Article  CAS  Google Scholar 

  • Nakamura SI, Suzui N, Yin YG, Ishii S, Fujimaki S, Kawachi N, Rai H, Matsumoto T, Sato-Izawa K, Ohkama-Ohtsu N (2020) Effects of enhancing endogenous and exogenous glutathione in roots on cadmium movement in Arabidopsis thaliana. Plant Sci 290:110304

    Article  CAS  Google Scholar 

  • Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate–specific peroxidase in spinach chloroplasts. Plant Cell Physiol 22:867–880

    CAS  Google Scholar 

  • Nazir MM, Noman M, Ahmed T, Ali S, Ulhassan Z, Zeng F, Zhang G (2022) Exogenous calcium oxide nanoparticles alleviate cadmium toxicity by reducing Cd uptake and enhancing antioxidative capacity in barley seedlings. J Hazard Mater 438:129498

    Article  CAS  Google Scholar 

  • Rao G, Ashraf U, Huang S, Cheng S, Abrar M, Mo Z, Pan S, Tang X (2018) Ultrasonic seed treatment improved physiological and yield traits of rice under lead toxicity. Environ Sci Pollut Res 25:33637–33644

    Article  CAS  Google Scholar 

  • RoyChoudhury A, Roy C, Sengupta DN (2007) Transgenic tobacco plants overexpressing the heterologous lea gene Rab16A from rice during high salt and water deficit display enhanced tolerance to salinity stress. Plant Cell Rep 26:1839–1859

    Article  CAS  Google Scholar 

  • Ruegsegger A, Brunold CH (1992) Effect of cadmium and/or removal of € kernels or shoots on the levels of cysteine, c-glutamylcysteine, glutathione, and TCA-soluble thiols in maize seedlings. Phyton 32:109–112

    CAS  Google Scholar 

  • Salavati J, Fallah H, Niknejad Y, Barari Tari D (2021) Methyl jasmonate ameliorates lead toxicity in Oryza sativa by modulating chlorophyll metabolism, antioxidative capacity and metal translocation. Physiol Mol Biol Plants 27(5):1089–1104

    Article  CAS  Google Scholar 

  • Shabaan M, Asghar HN, Akhtar MJ, Ali Q, Ejaz M (2021) Role of plant growth promoting rhizobacteria in the alleviation of lead toxicity to Pisum sativum L. Int J Phytoremediation 23(8):837–845

    Article  CAS  Google Scholar 

  • Shee R, Ghosh S, Khan P, Sahid S, Roy C, Shee D, Paul S, Datta R (2022) Glutathione regulates transcriptional activation of iron transporters via S-nitrosylation of bHLH factors to modulate subcellular iron homoeostasis. Plant Cell Environ 45:2176–2190

    Article  CAS  Google Scholar 

  • Singh S, Prasad SM (2019) Regulation of chromium toxicity tolerance in tomato and brinjal by calcium and sulfur through nitric oxide: involvement of enzymes of sulfur assimilation and the ascorbate-glutathione cycle. Environ Exp Bot 166:103789

    Article  CAS  Google Scholar 

  • Singh R, Parihar P, Prasad SM (2020) Sulphur and calcium attenuate arsenic toxicity in Brassica by adjusting ascorbate–glutathione cycle and Sulphur metabolism. Plant Growth Regul 91(2):221–235

    Article  CAS  Google Scholar 

  • Su L, Xie Y, He Z, Zhou X, Liu Y, Zhang R, Li C (2022) Selenium mitigates Cd-induced oxidative stress and photosynthesis inhibition in two cherry tomato cultivars. J Plant Nutr Soil Sci 22(3):3212–3227

    Article  CAS  Google Scholar 

  • Tauqeer HM, Hussain S, Abbas F, Iqbal M (2019) The potential of an energy crop “Conocarpus erectus” for lead phytoextraction and phytostabilization of chromium, nickel, and cadmium: An excellent option for the management of multi-metal contaminated soils. Ecotoxicol Environ Saf 173:273–284

    Article  CAS  Google Scholar 

  • Velikova V, Yordanov I, Edreva A (2000) Oxidative stress and some antioxidant systems in acid rain-treated bean plants. Plant Sci 151:59–66

    Article  CAS  Google Scholar 

  • Verma N, Pandey A, Tiwari S, Prasad SM (2021) Calcium mediated nitric oxide responses: Acquisition of nickel stress tolerance in cyanobacterium Nostoc muscorum ATCC 27893. Biochem Biophys Rep 26:100953

    CAS  Google Scholar 

  • Wang K, He J, Gao Y, Han K, Liu J, Wang Y (2022) Exogenous melatonin improved the growth and development of naked oat seedlings under cadmium stress. Environ Sci Pollut Res 1–10. https://doi.org/10.1007/s11356-022-21798-3

  • Wen K, Li X, Huang R, Nian H (2022) Application of exogenous glutathione decreases chromium translocation and alleviates its toxicity in soybean (Glycine max L.). Ecotoxicol Environ Saf 234:113405

    Article  CAS  Google Scholar 

  • Yao M, Ge W, Zhou Q, Zhou X, Luo M, Zhao Y, Wei B, Ji S (2021) Exogenous glutathione alleviates chilling injury in postharvest bell pepper by modulating the ascorbate-glutathione (AsA-GSH) cycle. Food Chem 352:129458

    Article  CAS  Google Scholar 

  • Yuan H, Guo Z, Liu Q, Gu C, Yang Y, Zhang Y, Dhankher OP, Huang S (2018) Exogenous glutathione increased lead uptake and accumulation in Iris lactea var. chinensis exposed to excess lead. Int J Phytoremediation 20(11):1136–1143

    Article  CAS  Google Scholar 

  • Zhou WJ, Leul M (1998) Uniconazole-induced alleviation of freezing injury in relation to changes in hormonal balance, enzyme activities and lipid peroxidation in winter rape. Plant Growth Regul 26:41–47

    Article  CAS  Google Scholar 

Download references

Funding

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Author information

Authors and Affiliations

  1. Department of Biology and Biochemistry, Science Faculty, Shahr-E-Qods Branch, Islamic Azad University, Tehran, Iran

    Shima Hanife

  2. Department of Biology, Faculty of Sciences, Central Tehran Branch, Islamic Azad University, Tehran, Iran

    Shahram Namdjoyan

  3. Department of Biorefinery Engineering, Faculty of New Technologies and Engineering, Shahid Beheshti University, Tehran, Iran

    Hossein Kermanian

Authors
  1. Shima Hanife
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shahram Namdjoyan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hossein Kermanian
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Shima Hanife: Conceptualization, investigation and writing of the manuscript. Shahram Namdjoyan: Investigation, supervision, analysis, and interpretation of data for the work. Hossein Kermanian: investigation, writing, reviewing and editing of the manuscript.

Corresponding author

Correspondence to Shahram Namdjoyan.

Ethics declarations

Ethics approval and consent to participate

Not applicable.

Consent to participate

Not applicable.

Consent to publish

Not applicable.

Competing interests

This paper’s authors declare that they do not have any contradictory financial interest or personal relationship that might have influenced their work.

Additional information

Responsible Editor: Gangrong Shi

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Hanife, S., Namdjoyan, S. & Kermanian, H. Synergistic effects of exogenous glutathione and calcium on ascorbate–glutathione cycle and glutathione-associated enzymes upregulation under lead stress in Brassica napus L. Environ Sci Pollut Res 30, 108979–108991 (2023). https://doi.org/10.1007/s11356-023-30000-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11356-023-30000-1

Keywords

Search

Navigation