Dimethylthiourea antagonizes oxidative responses by up-regulating expressions of pyrroline-5-carboxylate synthetase and antioxidant genes under arsenic stress

Abstract

Dimethylthiourea is an important plant growth regulator that mediates various physiological and metabolic processes of the plants. In the present study, role of dimethylthiourea in conferring arsenic stress tolerance to Cajanus cajan L. was investigated. Exposure to arsenic resulted in oxidative damage as evidenced by decreased germination percentage, radicle length, biomass accumulation, membrane stability index, protein, glyoxalase I and II, and antioxidants, together with enhanced cell death, reactive oxygen species, lipid peroxidation and activity of lipoxygenase in C. cajan L. However, exogenous application of dimethylthiourea along with arsenic decreased the reactive oxygen species and lipoxygenase activity, while increased the membrane stability index and antioxidants activities. Moreover, dimethylthiourea also up-regulated the glyoxalase (I and II) activity and the gene expression of antioxidants under arsenic stress. Also, dimethylthiourea application reduced the arsenic content than that measured in arsenic alone treated samples. Interestingly, treatment of dimethylthiourea uplifted the contents of ascorbic acid and glutathione, together with proline via up-regulating the activity and gene expression of pyrroline-5-carboxylate synthetase, one of the chief enzymes of its biosynthetic pathway. Enlisted findings suggested that dimethylthiourea can improve plant resistance to arsenic toxicity by regulating the gene expression of antioxidants and proline biosynthesizing enzymes, thereby reducing reactive oxygen species, lipid peroxidation and arsenic accumulation.

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References

  1. Abbas G, Murtaza B, Bibi I, Shahid M, Niazi NK, Khan MI, Amjad M, Hussain M, Natasha (2018) Arsenic uptake, toxicity, detoxification, and speciation in plants: physiological, biochemical, and molecular aspects. Int J Environ Res Public Health 15:59–103

    Article  Google Scholar 

  2. Anjum SA, Tanveer M, Hussain S, Shahzad B, Ashraf U, Fahad S, Hassan W, Jan S, Khan I, Saleem MF, Bajwa AA, Wang L, Mahmood A, Samad RA, Tung SA (2016) Osmoregulation and antioxidant production in maize under combined cadmium and arsenic stress. Environ Sci Pollut Res 23:11864–11875

    CAS  Article  Google Scholar 

  3. Bates LS, Waldren RP, Teare ID (1973) Rapid determination of free proline for water stress studies. Plant Soil 39:205–207

    CAS  Article  Google Scholar 

  4. 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

    CAS  Google Scholar 

  5. Chance M, Maehly AC (1955) Assay of catalases and peroxidases. Methods Enzymol 2:764–817

    Article  Google Scholar 

  6. Chandrakar V, Keshavkant S (2018) Nitric oxide and dimethylthiourea up-regulates pyrroline-5-carboxylate synthetase expression to improve arsenic tolerance in Glycine max L. Environ Prog Sustain Energy. https://doi.org/10.1002/ep.12978

    Article  Google Scholar 

  7. Chandrakar V, Dubey A, Keshavkant S (2016) Modulation of antioxidant enzymes by salicylic acid in arsenic exposed Glycine max L. J Soil Sci Plant Nutr 16:662–676

    CAS  Google Scholar 

  8. Chandrakar V, Yadu B, Meena R, Dubey A, Keshavkant S (2017a) Arsenic-induced genotoxic responses and their amelioration by diphenylene iodonium, 24-epibrassinolide and proline in Glycine max L. Plant Physiol Biochem 112:74–86

    CAS  Article  Google Scholar 

  9. Chandrakar V, Parkhey S, Dubey A, Keshavkant S (2017b) Modulation in arsenic induced lipid catabolism in Glycine Max using proline, 24-epibrassinolide and diphenylene iodonium. Biologia 72:292–299

    CAS  Article  Google Scholar 

  10. Chandrakar V, Dubey A, Keshavkant S (2018) Modulation of arsenic-induced oxidative stress and protein metabolism by diphenyleneiodonium, 24-epibrassinolide and proline in Glycine max L. Acta Bot Croat 77:51–61

    CAS  Article  Google Scholar 

  11. Farooq MA, Li L, Ali B, Gill RA, Wang J, Ali S, Gill MB, Zhou W (2015) Oxidative injury and antioxidant enzymes regulation in arsenic-exposed seedlings of four Brassica napus L. cultivars. Environ Sci Pollut Res 22:10699–10712

    CAS  Article  Google Scholar 

  12. Garcia-Rios M, Fujita T, Larosa PC, Locyi RD, Clithero JM, Bressan RA, Csonka LN (1997) Cloning of a poly cistronic cDNA from tomato encoding γ-glutamyl kinase and γ-glutamylphosphate reductase. Proc Natl Acad Sci USA 94:8249–8254

    CAS  Article  Google Scholar 

  13. Griffith OW (1980) Determination of glutathione and glutathione disulfide using glutathione reductase and 2-vinylpyridine. Anal Biochem 106:207–212

    CAS  Article  Google Scholar 

  14. Hasanuzzaman M, Fujita M (2013) Exogenous sodium nitroprusside alleviates arsenic-induced oxidative stress in wheat (Triticum aestivum L.) seedlings by enhancing antioxidant defence and glyoxalase system. Ecotoxicology 22:584–596

    CAS  Article  Google Scholar 

  15. Hodges DM, DeLong JM, Forney CF, Prange RK (1999) Improving the thiobarbituric acid-reactive-substances assay for estimating lipid peroxidation in plant tissues containing anthocyanin and other interfering compounds. Planta 207:604–611

    CAS  Article  Google Scholar 

  16. Hossain MA, Fujita M (2009) Purification of glyoxalase I from onion bulbs and molecular cloning of its cDNA. Biosci Biotechnol Biochem 73:2007–2013

    CAS  Article  Google Scholar 

  17. 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. J Exp Bot 53:2401–2410

    CAS  Article  Google Scholar 

  18. Kumari A, Pandey N, Pandey-Rai S (2018) Exogenous salicylic acid-mediated modulation of arsenic stress tolerance with enhanced accumulation of secondary metabolites and improved size of glandular trichomes in Artemisia annua L. Protoplasma 255:139–152

    CAS  Article  Google Scholar 

  19. Li CX, Feng SL, Shao Y, Jiang LN, Lu XY, Hou XL (2007) Effects of arsenic on seed germination and physiological activities of wheat seedlings. J Environ Sci 19:725–732

    CAS  Article  Google Scholar 

  20. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-δCT method. Methods 25:402–408

    CAS  Article  Google Scholar 

  21. Mallick S, Kumar N, Singh AP, Sinam G, Yadav RN, Sinha S (2013) Role of sulfate in detoxification of arsenate-induced toxicity in Zea mays L. (SRHM 445): nutrient status and antioxidants. J Plant Int 8:140–154

    CAS  Google Scholar 

  22. Marklund S, Marklund G (1974) Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 47:469–474

    CAS  Article  Google Scholar 

  23. Mukherjee SP, Choudhuri MA (1983) Implications of water stress-induced changes in the levels of endogenous ascorbic acid and hydrogen peroxide in Vigna seedlings. Physiol Plant 58:166–170

    CAS  Article  Google Scholar 

  24. Nair PMG, Chung IM (2015) Physiological and molecular level studies on the toxicity of silver nanoparticles in germinating seedlings of mung bean (Vigna radiata L.). Acta Physiol Plant 37:1–11

    CAS  Article  Google Scholar 

  25. Panda SK, Matsumoto H (2010) Changes in antioxidant gene expression and induction of oxidative stress in pea (Pisum sativum L.) under Al stress. Biometals 23:753–762

    CAS  Article  Google Scholar 

  26. Principato GB, Rosi G, Talesa V, Govannini E, Uolila L (1987) Purification and characterization of two forms of glyoxalase II from rat liver and brain of Wistar rats. Biochim Biophys Acta 911:349–355

    CAS  Article  Google Scholar 

  27. Sangeetha P, Das VN, Koratkar R, Suryaprabha P (1990) Increase in free radical generation and lipid peroxidation following chemotherapy in patients with cancer. Free Radic Biol Med 8:15–19

    CAS  Article  Google Scholar 

  28. Sengupta D, Guha A, Reddy AR (2013) Interdependence of plant water status with photosynthetic performance and root defense responses in Vigna radiata (L.) Wilczek under progressive drought stress and recovery. J Photochem Photobiol 127:170–181

    CAS  Article  Google Scholar 

  29. Srivastava AK, Sablok G, Hackenberg M, Deshpande U, Suprasanna P (2017) Thiourea priming enhances salt tolerance through co-ordinated regulation of microRNAs and hormones in Brassica juncea. Sci Rep 7:45490

    CAS  Article  Google Scholar 

  30. Talukdar D (2014) Arsenic-induced oxidative stress and its reversal by thiourea in mung bean (Vigna radiata L.) wilczek genotype. Cent Eur J Exp Biol 3:13–18

    Google Scholar 

  31. Talukdar D (2016) Exogenous thiourea modulates antioxidants defense and glyoxalase systems in lentil genotypes under arsenic stress. J Plant Stress Physiol 2:9–21

    Article  Google Scholar 

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

    CAS  Article  Google Scholar 

  33. Woodbury W, Spencer AK, Stahmann MA (1971) An improved procedure using ferricyanide for detecting catalase isozymes. Anal Biochem 44:301–305

    CAS  Article  Google Scholar 

  34. Xalxo R, Keshavkant S (2017) Acid rain-induced oxidative stress regulated metabolic interventions and their amelioration mechanisms in plants. Biologia 72:1387–1393

    CAS  Article  Google Scholar 

  35. Xalxo R, Yadu B, Chakraborty P, Chandrakar V, Keshavakant S (2017) Modulation of nickel toxicity by glycinebetaine and aspirin in Pennisetum typhoideum. Acta Biol Szeged 61:163–171

    Google Scholar 

  36. Yadu B, Chandrakar V, Meena RK, Keshavkant S (2017a) Glycinebetaine reduces oxidative injury and enhances fluoride stress tolerance via improving antioxidant enzymes, proline and genomic template stability in Cajanus cajan L. S Afr J Bot 111:68–75

    CAS  Article  Google Scholar 

  37. Yadu S, Dewangan TL, Chandrakar V, Keshavkant S (2017b) Imperative roles of salicylic acid and nitric oxide in improving salinity tolerance in Pisum sativum L. Physiol Mol Biol Plants 23:43–58

    CAS  Article  Google Scholar 

  38. Yadu B, Chandrakar V, Meena RK, Dubey A, Poddar A, Keshavkant S (2018a) Spermidine and melatonin attenuate fluoride toxicity by regulating gene expression of antioxidants in Cajanus cajan L. J Plant Growth Regul. https://doi.org/10.1007/s00344-018-9786-y

    Article  Google Scholar 

  39. Yadu B, Chandrakar V, Korram J, Satnami ML, Kumar M, Keshavkant S (2018b) Silver nanoparticle modulates gene expressions, glyoxalase system and oxidative stress markers in fluoride stressed Cajanus cajan L. J Hazard Mater 353:44–52

    CAS  Article  Google Scholar 

  40. Zhang A, Jiang M, Zhang J, Tan M, Hu X (2006) Mitogen-activated protein kinase is involved in abscisic acid-induced antioxidant defense and acts downstream of reactive oxygen species production in leaves of maize plants. Plant Physiol 141:475–487

    CAS  Article  Google Scholar 

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Acknowledgements

The authors would also like to thank Defense Research and Development Organization, New Delhi (DRDE/TC/05414/Proj/TASK-220/16 Dated June 27, 2016) and Department of Science and Technology, New Delhi (No. DST/INSPIRE Fellowship/2013/791, dated 23.01.2013) for providing research facilities for this study. Authors are also grateful to Department of Science and Technology, New Delhi, for financial support through DST-FIST scheme (Sanction No. 2384/IFD/2014-15, dated 31.07.2014) sanctioned to the School of Studies in Biotechnology.

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Correspondence to S. Keshavkant.

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Editorial responsbility: M. Abbaspour.

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Yadu, B., Chandrakar, V., Tamboli, R. et al. Dimethylthiourea antagonizes oxidative responses by up-regulating expressions of pyrroline-5-carboxylate synthetase and antioxidant genes under arsenic stress. Int. J. Environ. Sci. Technol. 16, 8401–8410 (2019). https://doi.org/10.1007/s13762-019-02234-5

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Keywords

  • Cajanus cajan L.
  • Cell death
  • Gene expression
  • Glyoxalase system
  • Lipoxygenase
  • Proline
  • Reactive oxygen species