Skip to main navigation menu Skip to main content Skip to site footer

Research Articles

Early Access

Unravelling nitric oxide–hydrogen sulfide interplay in NaCl-mediated salinity resistance of moth bean [Vigna aconitifolia (Jacq.) Marechal]

DOI
https://doi.org/10.14719/pst.9890
Submitted
7 June 2025
Published
27-08-2025
Versions

Abstract

Soil salinity causes oxidative stress, ion imbalance, osmotic anomaly, nutrient imbalance, and changes in plant growth regulators. The study aims to investigate the salinity tolerance limit, involvement of nitric oxide (NO) and hydrogen sulfide (H2S), and effects of different concentrations of exogenously supplied sodium chloride (NaCl) (50, 100, 150, 200, and 250 mM) on growth, physiological and biochemical parameters of the Moth bean [Vigna aconitifolia (Jacq.) Marechal] seedlings. Moth bean, a neglected and underutilised legume crop in India. The experimental results showed that salinity led to decrease in the plant growth [64.8% shoot length (SL), 58.6% root length (RL), 97.5% secondary roots], 19.3% fresh weight (FW), 16.01% dry weight (DW), (25.6%) water content, photosynthetic pigments (58.2% chl a, 20% chl b, 47.7% total chl, and 63.9% carotenoids), 27.3% antioxidant contents, and 23.7% increased malondialdehyde (MDA) content. The endogenous generation of NO (4.9%) and H2S (58%) were reported under salinity stress compared to the control treatment. The osmolyte proline (184.2%) and enhanced activities of antioxidant enzymes, such as catalase (CAT) (25.22%), peroxidase (POD) (22.25%), superoxide dismutase (SOD) (17.63%), and polyphenol oxidase (PPO) (24.80%), up to 150 mM NaCl treatment, marking the tolerance limit of the moth bean. Principal component analysis (PCA) and correlation analysis revealed the antagonistic generation pattern of NO and H2S, as well as their relationships with antioxidant enzymes under stress conditions. This is the first report on NO and H2S generation and involvement in Moth bean grown under salinity stress. The information obtained can be applied to augment salinity stress and enhance crop productivity.

References

  1. 1. Wang D, Bai J, Wang W, Zhang G, Cui B, Liu X, et al. Comprehensive assessment of soil quality for different wetlands in a Chinese delta. Land Degrad Dev. 2018;29(10):3783–94. https://doi.org/10.1002/ldr.3086
  2. 2. Singh A. Soil salinity: A global threat to sustainable development. Soil Use Manag. 2022;38(1):39–67. https://doi.org/10.1111/sum.12772
  3. 3. Mazarji M, Bayero MT, Minkina T, Sushkova S, Mandzhieva S, Tereshchenko A, et al. Realizing United Nations sustainable development goals for greener remediation of heavy metals-contaminated soils by biochar: Emerging trends and future directions. Sustainability. 2021;13(24):13825. https://doi.org/10.3390/su132413825
  4. 4. Pitman MG, Läuchli A. Global impact of salinity and agricultural ecosystems. In: Läuchli A, Lüttge U, editors. Salinity: Environment - Plants - Molecules. Dordrecht: Springer; 2002. p. 3–20. https://doi.org/10.1007/0-306-48155-3_1
  5. 5. Eswar D, Karuppusamy R, Chellamuthu S. Drivers of soil salinity and their correlation with climate change. Curr Opin Environ Sustain. 2021;50:310–8. https://doi.org/10.1016/j.cosust.2020.10.015
  6. 6. Li X, Wang A, Wan W, Luo X, Zheng L, He G, et al. High salinity inhibits soil bacterial community mediating nitrogen cycling. Appl Environ Microbiol. 2021;87(21):e01366-21. https://doi.org/10.1128/AEM.01366-21
  7. 7. Kumar P, Sharma PK. Soil salinity and food security in India. Front Sustain Food Syst. 2020;4:533781. https://doi.org/10.3389/fsufs.2020.533781
  8. 8. Bhadkaria A, Narvekar DT, Gupta N, Khare A, Bhagyawant SS. Moth bean (Vigna aconitifolia (Jacq.) Maréchal) seeds: a review on nutritional properties and health benefits. Discover Food. 2022;2(1):18. https://doi.org/10.1007/s44187-022-00019-3
  9. 9. Kanishka RC, Gayacharan, Basavaraja T, Chandora R, Rana JC. Moth bean (Vigna aconitifolia): a minor legume with major potential to address global agricultural challenges. Front Plant Sci. 2023;14:1179547. https://doi.org/10.3389/fpls.2023.1179547
  10. 10. Sharma R, Kumar S, Mahla HR, Choudhary KB, Khandelwal V. Moth bean (Vigna aconitifolia (Jacq.) Maréchal). In: Potential Pulses: Genetic and Genomic Resources. CABI GB; 2024. p. 264–86. https://doi.org/10.1079/9781800624658.0014
  11. 11. Farooq M, Gogoi N, Hussain M, Barthakur S, Paul S, Bharadwaj N, et al. Effects, tolerance mechanisms and management of salt stress in grain legumes. Plant Physiol Biochem. 2017;118:199–217. https://doi.org/10.1016/j.plaphy.2017.06.020
  12. 12. Dawood MFA, Sofy MR, Mohamed HI, Sofy AR, Abdel-Kader HAA. Hydrogen sulfide modulates salinity stress in common bean plants by maintaining osmolytes and regulating nitric oxide levels and antioxidant enzyme expression. J Soil Sci Plant Nutr. 2022;22(3):3708–26. https://doi.org/10.1007/s42729-022-00921-w
  13. 13. Wang Y, Li L, Cui W, Xu S, Shen W, Wang R. Hydrogen sulfide enhances alfalfa (Medicago sativa) tolerance against salinity during seed germination by nitric oxide pathway. Plant Soil. 2012;351:107–19. https://doi.org/10.1007/s11104-011-0936-2
  14. 14. Egbichi I, Keyster M, Ludidi N. Effect of exogenous application of nitric oxide on salt stress responses of soybean. S Afr J Bot. 2014;90:131–6. https://doi.org/10.1016/j.sajb.2013.11.002
  15. 15. Lai D, Mao Y, Zhou H, Li F, Wu M, Zhang J, et al. Endogenous hydrogen sulfide enhances salt tolerance by coupling the reestablishment of redox homeostasis and preventing salt-induced K⁺ loss in seedlings of Medicago sativa. Plant Sci. 2014;225:117–29. https://doi.org/10.1016/j.plantsci.2014.06.006
  16. 16. Liu Y, Wu R, Wan Q, Xie G, Bi Y. Glucose-6-phosphate dehydrogenase plays a pivotal role in nitric oxide-involved defense against oxidative stress under salt stress in red kidney bean roots. Plant Cell Physiol. 2007;48(3):511–22. https://doi.org/10.1093/pcp/pcm020
  17. 17. Tyagi A, Sharma S, Ali S, Gaikwad K. Crosstalk between H₂S and NO: an emerging signalling pathway during waterlogging stress in legume crops. Plant Biol. 2022;24(4):576–86. https://doi.org/10.1111/plb.13319
  18. 18. Sharma NK, Kumawat N, Panwar PK. Evaluation of high-yielding varieties and demonstration of production technologies in moth bean at farmers’ fields in Western Rajasthan. Ann Arid Zone. 2015;54(1&2):51–3. https://doi.org/10.56093/aaz.v54i1%20&%202.62851
  19. 19. Sauer D, Burroughs R. Disinfection of seed surfaces with sodium hypochlorite. Phytopathology. 1986;76(7):745–9. https://doi.org/10.1094/Phyto-76-745
  20. 20. Sumithra K, Jutur P, Carmel BD, Reddy AR. Salinity-induced changes in two cultivars of Vigna radiata: responses of antioxidative and proline metabolism. Plant Growth Regul. 2006;50:11–22. https://doi.org/10.1007/s10725-006-9121-7
  21. 21. Weatherley PE. Studies in the water relations of the cotton plant. New Phytol. 1950;49(1):81–97. https://doi.org/10.1111/j.1469-8137.1950.tb05146.x
  22. 22. Wilkins D. The measurement of tolerance to edaphic factors by means of root growth. New Phytol. 1978;80(3):623–33. https://doi.org/10.1111/j.1469-8137.1978.tb01595.x
  23. 23. Hiscox JD, Israelstam GF. A method for the extraction of chlorophyll from leaf tissue without maceration. Can J Bot. 1979;57(12):1332–4. https://doi.org/10.1139/b79-163
  24. 24. Arnon DI. Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol. 1949;24(1):1–15. https://doi.org/10.1104/pp.24.1.1
  25. 25. Heath RL, Packer L. Photoperoxidation in isolated chloroplasts: I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys. 1968;125(1):189–98. https://doi.org/10.1016/0003-9861(68)90654-1
  26. 26. Shafi A, Zahoor I, Mushtaq U. Proline accumulation and oxidative stress: diverse roles and mechanism of tolerance and adaptation under salinity stress. In: Salt Stress, Microbes, and Plant Interactions: Mechanisms and Molecular Approaches: Volume 2. 2019. p. 269–300. https://doi.org/10.1007/978-981-13-8805-7_13
  27. 27. Bates LS, Waldren R, Teare I. Rapid determination of free proline for water-stress studies. Plant Soil. 1973;39:205–7. https://doi.org/10.1007/BF00018060
  28. 28. Brand-Williams W, Cuvelier ME, Berset C. Use of a free radical method to evaluate antioxidant activity. LWT Food Sci Technol. 1995;28(1):25–30. https://doi.org/10.1016/S0023-6438(95)80008-5
  29. 29. 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. https://doi.org/10.1016/0003-2697(76)90527-3
  30. 30. Aebi H. Catalase in vitro. Methods Enzymol. 1984;105:121–6. https://doi.org/10.1016/s0076-6879(84)05016-3
  31. 31. Chance B, Maehly AC. Assay of catalase and peroxidase. Methods Enzymol. 1955;2:764–75. http://dx.doi.org/10.1016/S0076-6879(55)02300-8
  32. 32. Dhindsa R, Plumb-Dhindsa P, Thorpe T. Leaf senescence and lipid peroxidation. J Exp Bot. 1981;32(1):93–101. https://doi.org/10.1093/jxb/32.1.93
  33. 33. Gill SS, Anjum NA, Gill R, Yadav S, Hasanuzzaman M, Fujita M, et al. Superoxide dismutase—mentor of abiotic stress tolerance in crop plants. Environ Sci Pollut Res. 2015;22(14):10375–94. https://doi.org/10.1007/s11356-015-4532-5
  34. 34. Koji H, Ayako W, Toumi S. Changes in phenoloxidase activities of the galls on leaves of Ulmus davidiana formed by Tetraneura fusiformis (Homoptera: Eriosomatidae). Appl Entomol Zool. 1997;32(2):365–71. https://doi.org/10.1303/aez.32.365
  35. 35. Cheng T, Shi J, Dong Y, Ma Y, Peng Y, Hu X, et al. 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. 2018;84(1):11–23. https://doi.org/10.1007/s10725-017-0316-x
  36. 36. Zhou B, Guo Z, Xing J, Huang B. Nitric oxide is involved in abscisic acid-induced antioxidant activities in Stylosanthes guianensis. J Exp Bot. 2005;56(422):3223–8. https://doi.org/10.1093/jxb/eri319
  37. 37. Parihar P, Singh S, Singh R, Singh VP, Prasad SM. Effect of salinity stress on plants and its tolerance strategies: a review. Environ Sci Pollut Res. 2015;22:4056–75. https://doi.org/10.1007/s11356-014-3739-1
  38. 38. Bailly C, Benamar A, Corbineau F, Côme D. Changes in malondialdehyde content and in superoxide dismutase, catalase and glutathione reductase activities in sunflower seeds as related to deterioration during accelerated aging. Physiol Plant. 1996;97(1):104–10. https://doi.org/10.1111/j.1399-3054.1996.tb00485.x
  39. 39. Shahid MA, Sarkhosh A, Khan N, Balal RM, Ali S, Rossi L, et al. Insights into the physiological and biochemical impacts of salt stress on plant growth and development. Agronomy. 2020;10(7):938. https://doi.org/10.3390/agronomy10070938
  40. 40. Khan MN, Siddiqui ZH, Naeem M, Abbas ZK, Ansari MW. Nitric oxide and hydrogen sulfide interactions in plants under adverse environmental conditions. In: Aftab T, Naeem M, editors. Emerging Plant Growth Regulators in Agriculture. Academic Press; 2022. p. 215–44. https://doi.org/10.1016/B978-0-323-91005-7.00015-1
  41. 41. Hamada A, El-Enany A. Effect of NaCl salinity on growth, pigment and mineral element contents, and gas exchange of broad bean and pea plants. Biol Plant. 1994;36:75–81. https://doi.org/10.1007/BF02921273
  42. 42. Florina F, Giancarla V, Cerasela P, Sofia P. The effect of salt stress on chlorophyll content in several Romanian tomato varieties. J Hortic For Biotechnol. 2013;17(1):363–7.
  43. 43. Gao ZW, Wang ZJ, Cui YH, Ul-haq M, Ding KJ, Xu B, et al. Salinity stress in cultivated plants: toxic impacts, tolerance mechanisms and mitigation strategies. Appl Ecol Environ Res. 2025;23(2). http://dx.doi.org/10.15666/aeer/2302_17831814
  44. 44. Gill SS, Tuteja N. Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem. 2010;48(12):909–30. https://doi.org/10.1016/j.plaphy.2010.08.016
  45. 45. Kukreja S, Nandwal A, Kumar N, Sharma S, Sharma S, Unvi V, et al. Plant water status, H₂O₂ scavenging enzymes, ethylene evolution and membrane integrity of Cicer arietinum roots as affected by salinity. Biol Plant. 2005;49:305–8. https://doi.org/10.1007/s10535-005-5308-4
  46. 46. Taranto F, Pasqualone A, Mangini G, Tripodi P, Miazzi MM, Pavan S, et al. Polyphenol oxidases in crops: biochemical, physiological and genetic aspects. Int J Mol Sci. 2017;18(2). https://doi.org/10.3390/ijms18020377
  47. 47. Li ZG. Analysis of some enzymes activities of hydrogen sulfide metabolism in plants. In: Cadenas E, Packer L, editors. Methods in Enzymology. Vol. 555. Academic Press; 2015. p. 253–69. https://doi.org/10.1016/bs.mie.2014.11.035
  48. 48. Crawford NM. Mechanisms for nitric oxide synthesis in plants. J Exp Bot. 2006;57(3):471–8. https://doi.org/10.1093/jxb/erj050
  49. 49. Maurya AK, Agarwal R, Gupta R. Unraveling the crosstalk among ethylene, nitric oxide, and polyamines in tailoring the abiotic stress resilience in plants. Stress Biol. 2025;5(1):20. https://doi.org/10.1007/s44154-024-00198-2
  50. 50. Bhuyan MB, Hasanuzzaman M, Parvin K, Mohsin SM, Al Mahmud J, Nahar K, et al. Nitric oxide and hydrogen sulfide: two intimate collaborators regulating plant defense against abiotic stress. Plant Growth Regul. 2020;90:409–24. https://doi.org/10.1007/s10725-020-00594-4

Downloads

Download data is not yet available.