Abstract
The present study investigates a possible mechanism of 24-Epibrassinolide (EBL) mediated alleviation of salinity stress in Brassica juncea L. seedlings. EBL (10–6 M and 10–8 M) pretreated seedlings were grown up to ten-days either in alone and/or combination with defined levels of salinity treatments (100 mM and 200 mM NaCl) under laboratory growth conditions. Results showed that salinity stress significantly (P ≤ 0.05) increased the Na+ accumulation in a dose-dependent manner, which aggravated the nutrient(s) homeostasis and pigment contents by inducing oxidative damage through elevated hydrogen peroxide, malondialdehyde, and methyl glyoxal production. EBL mediated alleviation of salinity stress was achieved through decreased nuclear damage and cell death, coupled with increased membrane stability index and nutrients accumulation. The positive roles of EBL were also reflected through maintenance of AsA/DHA and GSH/GSSG redox ratios’ that were maximally enhanced by 78% and 28%, respectively under 200 mM NaCl. EBL mediated up-regulation of γ-glutamyl cysteine synthetase activity increased the glutathione pool, which boosted the ascorbate–glutathione cycle and methyl glyoxal detoxification system. The nitrate reductase activity, endogenous nitric oxide and S-nitrosothiols pools were maximally increased by 45, 32, and 33% respectively in 200 mM NaCl treatment, over the salinity alone counterpart. Taken together, our findings suggested that EBL pretreatment alleviates salinity stress by regulating nitric oxide homeostasis, osmolytes synthesis, antioxidant defense systems, and increasing the nutrients and pigments contents in mustard seedlings under salinity stress. Therefore, seed-priming with EBL can be considered as an effective approach for improving the plant productivity under salinity stress.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abat JK, Deswal R (2009) Differential modulation of S-nitrosoproteome of Brassica juncea by low temperature: change in S-nitrosylation of Rubisco is responsible for the inactivation of its carboxylase activity. Proteomics 9(18):4368–4380. https://doi.org/10.1002/pmic.200800985
Abd Allah EF, Alqarawi AA, Hashem A, Wirth S, Egamberdieva D (2018) Regulatory roles of 24-epibrassinolide in tolerance of Acacia gerrardii Benth to salt stress. Bioengineered 9(1):61–71. https://doi.org/10.1080/21655979.2017.1297348
Ahanger MA, Mir RA, Alyemeni MN, Ahmad P (2020) Combined effects of brassinosteroid and kinetin mitigates salinity stress in tomato through the modulation of antioxidant and osmolyte metabolism. Plant Physiol Biochem 147:31–42. https://doi.org/10.1016/j.plaphy.2019.12.007
Ahmad P, Abd Allah EF, Alyemeni MN, Wijaya L, Alam P, Bhardwaj R, Siddique KH (2018) Exogenous application of calcium to 24-epibrassinosteroid pretreated tomato seedlings mitigates NaCl toxicity by modifying ascorbate-glutathione cycle and secondary metabolites. Sci Rep 8(1):1–15. https://doi.org/10.1038/s41598-018-31917-1
Alam P, Albalawi TH, Altalayan FH, Bakht MA, Ahanger MA, Raja V, Ashraf M, Ahmad P (2019) 24-Epibrassinolide (EBR) confers tolerance against NaCl stress in soybean plants by up regulating antioxidant system, ascorbate-glutathione cycle, and glyoxalase system. Biomolecules 9(11):640. https://doi.org/10.3390/biom9110640
Asgher M, Per TS, Masood A, Fatma M, Freschi L, Corpas FJ, Khan NA (2017) Nitric oxide signaling and its crosstalk with other plant growth regulators in plant responses to abiotic stress. Environ Sci Pollut Res Int 24(3):2273–2285. https://doi.org/10.1007/s11356-016-7947-8
Azhar N, Su N, Shabala L, Shabala S (2017) Exogenously applied 24-epibrassinolide (EBL) ameliorates detrimental effects of salinity by reducing K+ efflux via depolarization-activated K+ channels. Plant Cell Physiol 58(4):802–810. https://doi.org/10.1093/pcp/pcx026
Bates LS, Waldren RP, Teare ID (1973) Rapid determination of free proline for water-stress studies. Plant Soil 39:205–207. https://doi.org/10.1007/s11104-016-3043-6
Ben Azaiez FE, Ayadi S, Capasso G, Landi S, Paradisone V, Jallouli S, Hammami Z, Chamekh Z, Zouari I, Trifa Y, Esposito S (2020) Salt stress induces differentiated nitrogen uptake and antioxidant responses in two contrasting barley landraces from MENA region. Agronomy 10(9):1426. https://doi.org/10.3390/agronomy10091426
Cheng T, Shi J, Dong Y, Ma Y, Peng Y, Hu X, Chen J (2018) Hydrogen sulfide enhances poplar tolerance to high-temperature stress by increasing S-nitrosoglutathione reductase (GSNOR) activity and reducing reactive oxygen/nitrogen damage. Plant Growth Regul 84(1):11–23. https://doi.org/10.1007/s10725-017-0316-x
Cheng L, Li M, Min W, Wang M, Chen R, Wang W (2021) Optimal brassinosteroid levels are required for soybean growth and mineral nutrient homeostasis. Int J Mol Sci 22(16):8400. https://doi.org/10.3390/ijms22168400
Cui JX, Zhou YH, Ding JG, Xia XJ, Shi KAI, Chen SC, Asami T, Chen Z, Yu JQ (2011) Role of nitric oxide in hydrogen peroxide-dependent induction of abiotic stress tolerance by brassinosteroids in cucumber. Plant Cell Environ 34(2):347–358. https://doi.org/10.1111/j.1365-3040.2010.02248.x
Debouba M, Dguimi HM, Ghorbel M, Gouia H, Suzuki A (2013) Expression pattern of genes encoding nitrate and ammonium assimilating enzymes in Arabidopsis thaliana exposed to short term NaCl stress. J Plant Physiol 170(2):155–160. https://doi.org/10.1016/j.jplph.2012.09.011
Esposito S (2016) Nitrogen assimilation, abiotic stress and glucose 6-phosphate dehydrogenase: the full circle of reductants. Plants 5(2):24. https://doi.org/10.3390/plants5020024
Esposito S, Carfagna S, Massaro G, Vona V, Rigano VDM (2001) Glucose-6-phosphate dehydrogenase in barley roots: kinetic properties and localisation of the isoforms. Planta 212(4):627–634. https://doi.org/10.1007/s004250000443
Evelin H, Devi TS, Gupta S, Kapoor R (2019) Mitigation of salinity stress in plants by arbuscular mycorrhizal symbiosis: current understanding and new challenges. Front Plant Sci 10:470. https://doi.org/10.3389/fpls.2019.00470
Grieve CM, Grattan SR (1983) Rapid assay for determination of water soluble quaternary ammonium compounds. Plant Soil 70(2):303–307. https://doi.org/10.1007/BF02374789
Gupta P, Seth CS (2015) Nitric oxide donor sodium nitroprusside promotes seed germination and ameliorates adverse effects of salinity by enhancing the growth indices and photosynthetic traits in Brassica juncea L. cv. Varuna. Phytomorphology 65(3&4):156–163
Gupta P, Seth CS (2020) Interactive role of exogenous 24 Epibrassinolide and endogenous NO in Brassica juncea L. under salinity stress: evidence for NR-dependent NO biosynthesis. Nitric Oxide 97:33–47. https://doi.org/10.1016/j.niox.2020.01.014
Jan S, Noman A, Kaya C, Ashraf M, Alyemeni MN, Ahmad P (2020) 24-Epibrassinolide alleviates the injurious effects of Cr (VI) toxicity in tomato plants: insights into growth, physio-biochemical attributes, antioxidant activity and regulation of ascorbate-glutathione and glyoxalase cycles. J Plant Growth Regul 39(4):1587–1604. https://doi.org/10.1007/s00344-020-10169-2
Jan S, Alyemeni MN, Wijaya L, Alam P, Siddique KH, Ahmad P (2018) Interactive effect of 24-epibrassinolide and silicon alleviates cadmium stress via the modulation of antioxidant defence and glyoxalase systems and macronutrient content in Pisum sativum L. seedlings. BMC Plant Biol 18(1):146. https://doi.org/10.1186/s12870-018-1359-5
Jaworski EG (1971) Nitrate reductase assay in intact plant tissues. Biochem Biophys Res Commun 43(6):1274–1279. https://doi.org/10.1016/S0006-291X(71)80010-4
Jedelská T, Šmotková Kraiczová V, Berčíková L, Činčalová L, Luhová L, Petřivalský M (2019) Tomato root growth inhibition by salinity and cadmium is mediated by S-nitrosative modifications of ROS metabolic enzymes controlled by S-nitrosoglutathione reductase. Biomolecules 9(9):393. https://doi.org/10.3390/biom9090393
Jin J, Li K, Qin J, Yan L, Wang S, Zhang G, Wang X, Bi Y (2021) The response mechanism to salt stress in Arabidopsis transgenic lines over-expressing of GmG6PD. Plant Physiol Biochem 162:74–85. https://doi.org/10.1016/j.plaphy.2021.02.021
Kaur H, Sirhindi G, Bhardwaj R, Alyemeni MN, Siddique KH, Ahmad P (2018) 28-homobrassinolide regulates antioxidant enzyme activities and gene expression in response to salt-and temperature-induced oxidative stress in Brassica juncea. Sci Rep 8(1):1–13. https://doi.org/10.1038/s41598-018-27032-w
Kaya C, Ashraf M, Alyemeni MN, Ahmad P (2020) Nitrate reductase rather than nitric oxide synthase activity is involved in 24-epibrassinolide-induced nitric oxide synthesis to improve tolerance to iron deficiency in strawberry (Fragaria× annassa) by up regulating the ascorbate-glutathione cycle. Plant Physiol Biochem 151:486–499. https://doi.org/10.1016/j.plaphy.2020.04.002
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 defence responses, and gene expression in Brassica juncea L. seedlings under Pb stress. Environ Sci Pollut Res 25(15):15159–15173. https://doi.org/10.1007/s11356-018-1742-7
Kohli SK, Bali S, Tejpal R, Bhalla V, Verma V, Bhardwaj R, Alqarawi AA, AbdAllah EF, Ahmad P (2019) In-situ localization and biochemical analysis of bio-molecules reveals Pb-stress amelioration in Brassica juncea L. by co-application of 24-Epibrassinolide and salicylic acid. Sci Rep 9(1):1–15. https://doi.org/10.1038/s41598-019-39712-2
Lawrence JF (1990) Determination of total xanthophyll and marigold oleoresin. J Assoc off Anal Chem 2:970–975
Li BB, Fu YS, Li XX, Yin HN, Xi ZM (2022) Brassinosteroids alleviate cadmium phytotoxicity by minimizing oxidative stress in grape seedlings: toward regulating the ascorbate-glutathione cycle. Sci Hortic 299:111002. https://doi.org/10.1016/j.scienta.2022.111002
Liu Y, Wu R, Wan Q, Xie G, Bi Y (2007) Glucose-6-phosphate dehydrogenase plays a pivotal role in nitric oxide-involved defence against oxidative stress under salt stress in red kidney bean roots. Plant Cell Physiol 48(3):511–522. https://doi.org/10.1093/pcp/pcm020
Ma Y, Wang L, Zhang W (2022) The role of hydrogen sulfide and its relationship with hydrogen peroxide and nitric oxide in brassinosteroid-induced stomatal closure of Vicia faba L. S Afr J Bot 146:426–436. https://doi.org/10.1016/j.sajb.2021.11.012
Munns R, Tester M (2008) Mechanisms of salinity tolerance. Annu Rev Plant Biol 59:651–681. https://doi.org/10.1146/annurev.arplant.59.032607.092911
Noctor G, Mhamdi A, Foyer CH (2016) Oxidative stress and antioxidative systems: recipes for successful data collection and interpretation. Plant Cell Environ 39(5):1140–1160. https://doi.org/10.1111/pce.12726
Rattan A, Kapoor D, Kapoor N, Bhardwaj R (2014) Application of brassionsteroids reverses the inhibitory effect of salt stress on growth and photosynthetic activity of Zea mays plants. IJTAS 6(2):13
Rüegsegger A, Brunold C (1992) Effect of cadmium on γ-glutamylcysteine synthesis in maize seedlings. Plant Physiol 99(2):428–433. https://doi.org/10.1104/pp.99.2.428
Sakamoto A, Ueda M, Morikawa H (2002) Arabidopsis glutathione-dependent formaldehyde dehydrogenase is an S-nitrosoglutathione reductase. FEBS Lett 515(1–3):20–24. https://doi.org/10.1016/S0014-5793(02)02414-6
Sharma A, Thakur S, Kumar V, Kanwar MK, Kesavan AK, Thukral AK, Bhardwaj R, Alam P, Ahmad P (2016) Pre-sowing seed treatment with 24-epibrassinolide ameliorates pesticide stress in Brassica juncea L. through the modulation of stress markers. Front Plant Sci 7:1569. https://doi.org/10.3389/fpls.2016.01569
Su Q, Zheng X, Tian Y, Wang C (2020) Exogenous brassinolide alleviates salt stress in Malus hupehensis rehd. by regulating the transcription of NHX-Type Na+(K+)/H+ antiporters. Front Plant Sci 11:38. https://doi.org/10.3389/fpls.2020.00038
Wahid I, Rani P, Kumari S, Ahmad R, Hussain SJ, Alamri S, Tripathy N, Khan MIR (2022) Biosynthesized gold nanoparticles maintained nitrogen metabolism, nitric oxide synthesis, ions balance, and stabilizes the defense systems to improve salt stress tolerance in wheat. Chemosphere 287:132142. https://doi.org/10.1016/j.chemosphere.2021.132142
Wu QS, Zou YN, Xia RX (2006) Effects of water stress and arbuscular mycorrhizal fungi on reactive oxygen metabolism and antioxidant production by citrus (Citrus tangerine) roots. Eur J Soil Biol 42(3):166–172. https://doi.org/10.1016/j.ejsobi.2005.12.006
Wu XX, Chen JL, Xu S, Zhu ZW, Zha DS (2016) Exogenous 24-epibrassinolide alleviates zinc-induced toxicity in eggplant (Solanum melongena L) seedlings by regulating the glutathione-ascorbate-dependent detoxification pathway. Hortic Sci Biotechnol 91(4):412–420. https://doi.org/10.1080/14620316.2016.1162030
Yin YL, Zhou Y, Zhou YH, Shi K, Zhou J, Yu Y, Yu JQ, Xia XJ (2016) Interplay between mitogen-activated protein kinase and nitric oxide in brassinosteroid-induced pesticide metabolism in Solanum lycopersicum. J Hazard Mater 316:221–231. https://doi.org/10.1016/j.jhazmat.2016.04.070
Yuan LB, Peng ZH, Zhi TT, Zho Z, Liu Y, Zhu Q, Xiong XY, Ren CM (2015) Brassinosteroid enhances cytokinin-induced anthocyanin biosynthesis in Arabidopsis seedlings. Bio Plant 59(1):99–105. https://doi.org/10.1007/s10535-014-0472-z
Zhou Y, Wen Z, Zhang J, Chen X, Cui J, Xu W, Liu HY (2017) Exogenous glutathione alleviates salt-induced oxidative stress in tomato seedlings by regulating glutathione metabolism, redox status, and the antioxidant system. Sci Hortic 220:90–101. https://doi.org/10.1016/j.scienta.2017.02.021
Zhu T, Deng XG, Tan WR, Zhou X, Luo SS, Han XY, Zhang DW, Lin HH (2016) Nitric oxide is involved in brassinosteroid induced alternative respiratory pathway in Nicotiana benthamiana seedlings’ response to salt stress. Physiol Plant 156(2):150–163. https://doi.org/10.1111/ppl.12392
Zou LJ, Deng XG, Zhang LE, Zhu T, Tan WR, Muhammad A, Zhu LJ, Zhang C, Zhang DW, Lin HH (2018) Nitric oxide as a signalling molecule in brassinosteroid-mediated virus resistance to Cucumber mosaic virus in Arabidopsis thaliana. Physiol Plant 163(2):196–210. https://doi.org/10.1111/ppl.12677
Acknowledgements
Chandra Shekhar Seth is thankful to Department of Botany, University of Delhi for providing laboratory facility and Institution of Eminence (IoE), University of Delhi is acknowledged for financial assistance vide Ref No. IoE/2021/12/FRP.
Author information
Authors and Affiliations
Contributions
(1) Conceptualization and experimental design: PG, CSS. (2) Methodology, data collection, and data analysis: PG. (3) Funding acquisition: CSS. (4) Intellectual inputs and manuscript writing: PG, CSS. (5) Final approval: CSS.
Corresponding author
Ethics declarations
Conflict of interest
On behalf of all authors, the corresponding author states that there is no conflict of interest.
Additional information
Handling Editor: Durgesh Kumar Tripathi.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
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
Gupta, P., Seth, C.S. 24-Epibrassinolide Regulates Functional Components of Nitric Oxide Signalling and Antioxidant Defense Pathways to Alleviate Salinity Stress in Brassica juncea L. cv. Varuna. J Plant Growth Regul 42, 4207–4222 (2023). https://doi.org/10.1007/s00344-022-10884-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00344-022-10884-y