Amelioration of the adverse effects of thiram by 24-epibrassinolide in tomato (Solanum lycopersicum Mill.)




Fungicide, brassinosteroid, chlorophyll, oxidative stress, pesticide detoxification enzyme


Paper description:

  • Intensive of pesticides negatively affects plant growth and yield. There is insufficient information on the negative effects of the dithiocarbamate fungicide thiram which is widely used in agricultural applications, and how these effects can be improved.
  • The effects of the plant hormone 24-epibrassinolide on thiram induced-stress in tomatoes were investigated. Photosynthetic pigment contents, oxidative stress indicators, the activities of antioxidant enzymes, and pesticide detoxification enzymes were analyzed in tomato (Solanum lycopersicum).
  • 24-epibrassinolide significantly reduced the adverse effects of thiram.
  • 24-epibrassinolide application to plants against pesticide toxicity has the potential to improve crop yield and production.

Abstract: This study investigated thiram fungicide-induced-stress effects in tomato plants and the possible protective role of 24-epibrassinolide (24-EBL) in response to thiram (tetramethyl thiuram disulfide) toxicity. Tomato seedlings pretreated with 0, 10-11, 10-9 and 10-7 M 24-EBL were treated with 6.6 mM thiram. Tomato leaves harvested 5 and 11 days after thiram treatment (DAT) were used for analysis. Thiram application caused oxidative stress by increased hydrogen peroxide and malondialdehyde levels, whereas the chlorophyll a, b and carotenoid amounts and total protein content decreased. In addition, the activities of antioxidant enzymes such as catalase, ascorbate peroxidase and glutathione reductase decreased in the thiram-treated tomato plants on DAT 5 and 11 while pesticide detoxification enzymes (peroxidase and glutathione-S-transferase) activities increased. The thiram-induced oxidative stress was alleviated after pretreatments with different concentrations of 24-EBL. The hydrogen peroxide and malondialdehyde levels decreased and the amounts of photosynthetic pigments and total protein content increased after 24-EBL pretreatments. In addition, the activities of antioxidant enzymes and pesticide detoxification enzymes further increased as the concentration of 24-EBL decreased in tomato under thiram stress, and the most effective concentration was determined as 10-11 M 24-EBL. The results suggested that 24-EBL could effectively alleviate thiram-induced phytotoxicity in tomato plants.


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Parween T, Jan S, Mahmooduzzafar S, Fatma T, Siddiqui ZH. Selective effect of pesticides on plant—a review. Crit Rev Food Sci Nutr. 2016;56:160-79.

Carvalho FP. Agriculture, pesticides, food security and food safety. Environ Sci Policy. 2006;9:685-92.

Yüzbaşıoğlu E, Dalyan E. Salicylic acid alleviates thiram toxicity by modulating antioxidant enzyme capacity and pesticide detoxification systems in the tomato (Solanum lycopersicum Mill.). Plant Physiol Biochem. 2019;135:322-30.

Prevention, pesticides and toxic substances [Internet]. Washington: US Environmental Protection Agency; c2004 [cited 2023 Mar 15]. Available from:

Priyantha N, Navaratne A, Ekanayake CB, Ratnayake A. Solvent extraction followed by ultraviolet detection for investigation of tetramethylthiuram disulfide at soil-water interface. Int J Environ Sci Technol. 2008;5:547-54.

Chahrazad B, Ibtissem S. Assessing the effects of thiram to oxidative stress responses in a freshwater bioindicator cladoceran (Daphnia magna). Chemosphere. 2021;268:128808.

Zhou Y, Xia X, Yu G, Wang J, Wu J, Wang M, Yang Y, Shi K, Yu Y, Chen Z, Gan J, Yu J. Brassinosteroids play a critical role in the regulation of pesticide metabolism in crop plants. Sci Rep. 2015;5:e9018.

Sharma A, Kumar V, Kanwar MK, Thukral KA, Bhardwaj R. Ameliorating imidacloprid induced oxidative stress by 24-epibrassinolide in Brassica juncea L. Russ J Plant Physiol. 2017;64:509-17.

Sharma A, Yuan H, Kumar V, Ramakrishnan M, Kohli SK, Kaur R, Thukral AK, Bhardwaj R, Zheng B. Castasterone attenuates insecticide induced phytotoxicity in mustard. Ecotoxicol Environ Saf. 2019;179:50-61.

Sharma A, Kumar V, Yuan H, Kanwar MK, Bhardwaj R, Thukral AK, Zheng B. Jasmonic acid seed treatment stimulates insecticide detoxification in Brassica juncea L. Front Plant Sci. 2018;9:e1609.

Caverzan A, Casassola A, Brammer SP. Reactive oxygen species and antioxidant enzymes involved in plant tolerance to stress, abiotic and biotic stress in plants. In: Shanker AK, Shanker C, editors. Abiotic and biotic stress in plants—Recent advances and future perspectives, Rijeka: InTech; 2016. p. 463-80.

Coleman J, Blake-Kalff M, Davies E. Detoxification of xenobiotics by plants: chemical modification and vacuolar compartmentation. Trends Plant Sci. 1997;2:144-51.

Zhang JJ, Yang H. Metabolism and detoxification of pesticides in plants. Sci Total Environ. 2021;790:148034.

Choudhary SP, Yu J-Q, Yamaguchi-Shinozaki K, Shinozaki K, Tran LSP. Benefits of brassinosteroid crosstalk. Trends Plant Sci. 2012;17:594-605.

Baghel M, Nagaraja A, Srivastav M, Meena NK, Kumar MS, Kumar A, Sharma RR. Pleiotropic influences of brassinosteroids on fruit crops: a review. Plant Growth Regul. 2019:87;375-88.

Bajguz A, Hayat S. Effects of brassinosteroids on the plant responses to environmental stresses. Plant Physiol Biochem. 2009;47:1-8.

Manghwar H, Hussain A, Ali Q, Liu F. Brassinosteroids (BRs) role in plant development and coping with different stresses. Int J Mol Sci. 2022;23:1012.

Sharma A, Kumar V, Kumar R, Shahzad B, Thukral KA, Bhardwaj R. Brassinosteroid-mediated pesticide detoxification in plants: a mini-review. Cogn Food Agric. 2018;4:e1436212.

Sharma I, Bhardwaj R, Pati PK. Stress modulation response of 24-epibrassinolide against imidacloprid in an elite indica rice variety Pusa Basmati-1. Pestic Biochem Phys. 2013;105:144-53.

Wang Z, Jiang Y, Peng X, Xu S, Zhang H, Gao J, Xi Z. Exogenous 24-epibrassinolide regulates antioxidant and pesticide detoxification systems in grapevine after chlorothalonil treatment. Plant Growth Regul. 2017;81:455-66.

Xia XJ, Zhang Y, Wu JX, Wang JT, Zhou YH, Shi K, Yu YL, Yu JQ. Brassinosteroids promote metabolism of pesticides in cucumber. J Agr Food Chem. 2009;57:8406-13.

Sun LW, Ren L, Peng T, Xin LH, Qian HF, Fu ZW. Carbon dioxide enrichment and brassinosteroid pretreatment alleviate chlorpyrifos phytotoxicity under suboptimal light and temperature conditions in tomato. Sci Hortic. 2015;192:256-63.

Sharma A, Kumar V, Bhardwaj R, Thukral AK. Seed pre-soaking with 24-epibrassinolide reduces the imidacloprid pesticide residues in green pods of Brassica juncea L.. Toxicol Environ Chem. 2017;99:95-103.

Hoagland DR, Arnon DI. The water culture method for growing plants without soil. Circ Calif Agric Exp Stn. 1950;347:32.

Lichtenthaler HK, Buschmann C. Chlorophylls and carotenoids: measurement and characterization by UV-VIS spectroscopy. Curr Protoc Food Anal Chem. 2001;1:F4.3.1-F4.3.8.

Velikova V, Yordanov I, Edreva A. Oxidative stress and some antioxidant systems in acid rain-treated bean plants: protective role of exogenous polyamines. Plant Sci. 2000;151:59-66.

Jiang M, Zhang J. Effect of abscisic acid on active oxygen species, antioxidative defense system and oxidative damage in leaves of maize seedlings. Plant Cell Physiol. 2001;42:1265-73.

Bergmeyer HU. Methoden der Enzymatischen Analyse. Berlin: Verlag Chemie; 1970.

Nakano Y, Asada K. Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant Cell Physiol. 1981;22:867-80.

Foyer CH, Halliwell B. The presence of glutathione and glutathione reductase in chloroplast: a proposed role in ascorbic acid metabolism. Planta. 1976;133:21-5.

Herzog V, Fahimi H. A new sensitive colorimetric assay for peroxidase using 3,3′-diaminobenzidine as hydrogen donor. Anal Biochem. 1973;55:554-62.

Habig, WH, Jacoby WB. Assays for differentiation of glutathione S-transferases. Methods Enzymol. 1981;77:398-405.

Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248-54.

Petit AN, Fontaine F, Vatsa P, Clement C, Vaillant-Gaveau N. Fungicide impacts on photosynthesis in crop plants. Photosynth Res. 2012;111:315-26.

Singh G, Sahota HK. Impact of benzimidazole and dithiocarbamate fungicides on the photosynthetic machinery, sugar content and various antioxidative enzymes in chickpea. Plant Physiol Biochem. 2018;132:166-73.

Sharma A, Kumar V, Singh R, Thukral AK, Bhardwaj R. Effect of seed pre-soaking with 24-epibrassinolide on growth and photosynthetic parameters of Brassica juncea L. in imidacloprid soil. Ecotoxicol Environ Saf. 2016;133:195-201.

Hola D. Brassinosteroids and photosynthesis. In: Hayat S, Ahmad A, editors. Brassinosteroids: A Class of Plant Hormone. Springer; 2011. p. 185-92.

Bakshi P, Chouhan R, Sharma P, Mir BA, Gandhi SG, Landi M, Zheng B, Sharma A, Bhardwaj R. Amelioration of chlorpyrifos-induced toxicity in Brassica juncea L. by combination of 24-epibrassinolide and plant-growth-promoting rhizobacteria. Biomolecules. 2021;11:e877.

Vardhini BV, Anjum NA. Brassinosteroids make plant life easier under abiotic stresses mainly by modulating major components of antioxidant defence system. Front Enviro. Sci. 2015;2:e67.

Dalyan E. 24-Epibrassinolide mitigates Cd-induced toxicity through improved antioxidative system In Maize (Zea mays L.) plant. Fresenius Environ Bull. 2020;29(09A):8421-34.

Sharma A, Thakur S, Kumar V, Kesavan AK, Thukral AK, Bhardwaj R. 24-epibrassinolide stimulates imidacloprid detoxification by modulating the gene expression of Brassica juncea L.. BMC Plant Biol. 2017;17:e56.

Wang J, Zhong X, Li F, Shi Z. Effects of nicosulfuron on growth, oxidative damage, and the ascorbate-glutathione pathway in paired nearly isogenic lines of waxy maize (Zea mays L.). Pestic Biochem Physiol. 2018;145:108-17.

Dalyan E, Yüzbaşıoğlu E, Akpınar I. Effect of 24-epibrassinolide on antioxidative defence system against lead-induced oxidative stress in the roots of Brassica juncea L. seedlings. Russ J Plant Physiol. 2018;65:570-8.

Liu S, He Y, Tian H, Yu C, Tan W, Li Z, Duan L. Application of brassinosteroid mimetics improves growth and tolerance of maize to nicosulfuron toxicity. Plant Growth Regul. 2019;38:701-12.

Sharma A, Kumar V, Thukral KA, Bhardwaj R. 24-epibrassinolide restores the synthesis of proteins and amino acids in Brassica juncea L. leaves under imidacloprid stress. J Horticult Res. 2017;25:85-90.

Sharma I, Bhardwaj R, Pati PK. Mitigation of adverse effects of chlorpyrifos by 24-epibrassinolide and analysis of stress markers in a rice variety Pusa Basmati-1. Ecotoxicol Environ Saf. 2012;85:72-81.

Dhaubhadel S, Browning KS, Gallie DR, Krishna P. Brassinosteroid functions to protect the translational machinery and heat-shock protein synthesis following thermal stress. Plant J. 2002;29:681-91.

Jan S, Singh R, Bhardwaj R, Ahmad P, Kapoor D. Plant growth regulators: a sustainable approach to combat pesticide toxicity. 3 Biotech. 2020;10:e466.

Farkas MH, Berry JO, Aga DS. Chlortetracycline detoxification in maize via induction of glutathione S-transferases after antibiotic exposure. Environ Sci Technol. 2007;41:1450-6.

Wang JT, Jiang YP, Chen SC, Xia XJ, Shi K, Zhou YH. The different responses of glutathione-dependent detoxification pathway to fungicide chlorothalonil and carbendazim in tomato leaves. Chemosphere. 2010;79:958-65.

Hao J, Yin Y, Fei S. Brassinosteroid signaling network: implications on yield and stress tolerance. Plant Cell Rep. 2013;32:1017-30.




How to Cite

Dalyan E. Amelioration of the adverse effects of thiram by 24-epibrassinolide in tomato (Solanum lycopersicum Mill.). Arch Biol Sci [Internet]. 2023Jul.3 [cited 2023Dec.10];75(2):187-9. Available from: