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

ONLINE FIRST

Research paper

Effect of cyclic 3-hydroxymelatonin (3-OHM) on pepper seedling emergence under drought conditions

DOI: https://doi.org/10.24326.asphc.2026.5575
Submitted: 24 July 2025
Published: 06.05.2026

Abstract

Cyclic 3-hydroxymelatonin (3-OHM), a significant metabolite derived from melatonin (MEL) through its interaction with oxygen-containing compounds, is believed to play a crucial role in enhancing plant resistance to various abiotic stresses. Despite its importance, research on 3-OHM remains limited. Therefore, this study aimed to investigate the effects of exogenous 3-OHM treatments on the drought stress tolerance of pepper seedlings during the emergence phase. The application of 3-OHM to seeds at various concentrations (0, 10, 50, and 100 µM) notably improved seedling emergence performance under drought conditions compared to untreated controls. Furthermore, 3-OHM treatments significantly reduced oxidative stress markers such as hydrogen peroxide (H₂O₂) and thiobarbituric acid reactive substances (TBARS), while simultaneously enhancing the activities of key antioxidant enzymes including peroxidase (POD), superoxide dismutase (SOD) and catalase (CAT). Additionally, seedling proline and total chlorophyll contents increased significantly by 3-OHM application. Among the concentrations tested, 50 µM 3-OHM consistently showed the most pronounced beneficial effects across multiple parameters. These results underscore the potential utility of 3-OHM as a natural bioactive compound to mitigate the detrimental impacts of abiotic stress in agricultural crops. To further elucidate the physiological mechanisms and confirm the efficacy of 3-OHM, future studies should focus on comparative analyses with MEL, employing the optimal 3-OHM concentration identified herein.

References

  1. Agarwal, S., Pandey, V. (2004). Antioxidant enzyme responses to NaCl stress in Cassia angustifolia. Biol. Plant., 48(4), 555‒560. https://doi.org/10.1023/B:BIOP.0000047152.07878.e7
  2. Ahmad, S., Ahmad, R., Ashraf, M.Y. et al. (2009). Sunflower (Helianthus annuus L.) response to drought stress at germination and seedling growth stages. Pak. J. Bot., 41(2), 647‒654.
  3. Álvarez-Diduk, R., Galano, A., Tan, D.X. et al. (2016). The key role of the sequential proton loss electron transfer mechanism on the free radical scavenging activity of some melatonin-related compounds. Theor Chem. Acc., 135(2), 38. https://doi.org/10.1007/s00214-015-1785-5
  4. Anjum, S.A., Ashraf, U., Zohaib, A. et al. (2017). Growth and development responses of crop plants under drought stress: a review. Zemdirbyste-Agriculture, 104(3), 267–276. http://dx.doi.org/10.13080/z-a.2017.104.034
  5. Back, K. (2021). Melatonin metabolism, signaling and possible roles in plants. Plant J., 105(2), 376‒391. https://doi.org/10.1111/tpj.14915
  6. Bates, L.S., Waldren, R.P.A., Teare, I.D. (1973). Rapid determination of free proline for water-stress studies. Plant Soil, 39, 205‒207. https://doi.org/10.1007/BF00018060
  7. Bewley, J.D. (1997). Seed germination and dormancy. Plant Cell, 9(7), 1055–1066. https://doi.org/10.1105/tpc.9.7.1055
  8. Bugg, T.D.H. (2003). Dioxygenase enzymes: catalytic mechanisms and chemical models. Tetrahedron, 59(36), 7075‒7101. https://doi.org/10.1016/S0040-4020(03)00944-X
  9. Bradford, M.M. (1976). A rapid and sensitive method for the quantitation 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
  10. Byeon, Y., Back, K. (2015). Molecular cloning of melatonin 2‐hydroxylase responsible for 2‐hydroxymelatonin production in rice (Oryza sativa). J. Pineal Res., 58, 343‒351. https://doi.org/10.1111/jpi.12220
  11. Byeon, Y., Tan, D.X., Reiter, R.J. et al. (2015). Predominance of 2‐hydroxymelatonin over melatonin in plants. J. Pineal Res. 59(4), 448‒454. https://doi.org/10.1111/jpi.12274
  12. Chazen, O., Hartung, W., Neumann, P.M. (1995). The different effects of PEG 6000 and NaCI on leaf development are asso-ciated with differential inhibition of root water transport. Plant. Cell Environ., 18(7), 727‒735. https://doi.org/10.1111/j.1365-3040.1995.tb00575.x
  13. Choi, G.H., Back, K. (2019a). Cyclic 3-hydroxymelatonin exhibits diurnal rhythm and cyclic 3-hydroxymelatonin overpro-duction increases secondary tillers in rice by upregulating MOC1 expression. Melatonin Res., 2(3), 120‒138. https://doi.org/10.32794/mr11250034
  14. Choi, G.H., Back, K. (2019b). Suppression of melatonin 2-hydroxylase increases melatonin production leading to the en-hanced abiotic stress tolerance against cadmium, senescence, salt, and tunicamycin in rice plants. Biomolecules, 9(10), 589. https://doi.org/10.3390/biom9100589
  15. Dolatabadian, A., Sanavy, S.M., Chashmi, N.A. (2008). The effects of foliar application of ascorbic acid (vitamin C) on antiox-idant enzymes activities, lipid peroxidation and proline accumulation of canola (Brassica napus L.) under conditions of salt stress. J. Agron. Crop Sci., 194(3), 206‒213. https://doi.org/10.1111/j.1439-037X.2008.00301.x
  16. Galano, A., Tan, D.X., Reiter, R.J. (2014). Cyclic 3-hydroxymelatonin, a key metabolite enhancing the peroxyl radical scav-enging activity of melatonin. RSC Adv., 4(10), 5220‒5227. https://doi.org/10.1039/C3RA44604B
  17. Gong, Y., Toivonen, P.M., Lau, O.L. et al. (2001). Antioxidant system level in 'Braeburn' apple is related to its browning dis-order. Bot. Bull. Acad. Sin., 42, 259–264
  18. Gunes, A., Inal, A., Bagci, E.G. et al. (2007). Silicon mediates changes to some physiological and enzymatic parameters symptomatic for oxidative stress in spinach (Spinacia oleracea L.) grown under B toxicity. Sci. Hortic., 113(2), 113‒119. https://doi.org/10.1016/j.scienta.2007.03.009
  19. Hajihashemi, S., Sofo, A. (2018). The effect of polyethylene glycol-induced drought stress on photosynthesis, carbohy-drates and cell membrane in Stevia rebaudiana grown in greenhouse. Acta Physiol. Plant., 40, 142. https://doi.org/10.1007/s11738-018-2722-8
  20. Hardeland, R. (2016). Melatonin in plants – diversity of levels and multiplicity of functions. Front. Plant Sci., 7, 198. https://doi.org/10.3389/fpls.2016.00198
  21. Hardeland, R. (2017). Taxon- and site-specific melatonin catabolism. Molecules, 22(11), 2015. https://doi.org/10.3390/molecules22112015
  22. Korkmaz, A, Sözeri, E., Ardıç, Ş.K. et al. (2023). 2-hydroxymelatonin (2-OHM), a major melatonin metabolite, confers multiple stress tolerance in pepper at seed germination stage. S. Afr. J. Bot., 162, 830‒837. https://doi.org/10.1016/j.sajb.2023.09.056
  23. Lee, K., Zawadzka, A., Czarnocki, Z. et al. (2016). Molecular cloning of melatonin 3‐hydroxylase and its production of cyclic 3‐hydroxymelatonin in rice (Oryza sativa). J. Pineal Res., 61(4), 470‒478. https://doi.org/10.1111/jpi.12361
  24. Lee, H.J., Back, K. (2016). 2‐Hydroxymelatonin promotes the resistance of rice plant to multiple simultaneous abiotic stresses (combined cold and drought). J. Pineal Res., 61(3), 303‒316.https://doi.org/10.1111/jpi.12347
  25. Lee, H.J., Back, K. (2019). 2-Hydroxymelatonin confers tolerance against combined cold and drought stress in tobacco, tomato, and cucumber as a potent anti-stress compound in the evolution of land plants. Melatonin Res., 2(2), 35‒46. http://dx.doi.org/10.32794/mr11250020
  26. Lee, H.Y, Back, K. (2022). The antioxidant cyclic 3-hydroxymelatonin promotes the growth and flowering of Arabidopsis thaliana. Antioxidants, 11(6), 1157. https://doi.org/10.3390/antiox11061157
  27. Ozden, M., Demirel, U., Kahraman A. (2009). Effects of proline on antioxidant system in leaves of grapevine (Vitis vinifera L.) exposed to oxidative stress by H2O2. Sci. Hortic., 119(2), 163‒168. https://doi.org/10.1016/j.scienta.2008.07.031
  28. Pérez-González, A., Galano, A., Alvarez-Idaboy, J.R. et al. (2017). Radical-trapping and preventive antioxidant effects of 2-hydroxymelatonin and 4-hydroxymelatonin: contributions to the melatonin protection against oxidative stress. BBA Gen. Subjects, 1861(9), 2206‒2217. https://doi.org/10.1016/j.bbagen.2017.06.016
  29. Posmyk, M.M., Bałabusta, M., Wieczorek, M. et al. (2009). Melatonin applied to cucumber (Cucumis sativus L.) seeds improves germination during chilling stress. J. Pineal Res., 46(2), 214‒223. https://doi.org/10.1111/j.1600-079X.2008.00652.x
  30. Rehaman, A., Mishra, A.K., Ferdose, A. et al. (2021). Melatonin in plant defense against abiotic stress. Forests, 12(10), 1404. https://doi.org/10.3390/f12101404
  31. Reiter, R.J., Mayo, J.C., Tan, D.X. et al. (2016). Melatonin as an antioxidant: under promises but over delivers. J. Pineal Res., 61(3), 253‒278. https://doi.org/10.1111/jpi.12360
  32. Shah, A.A, Ahmed, S., Ali, A. et al. (2020a). 2-Hydroxymelatonin mitigates cadmium stress in cucumis sativus seedlings: modulation of antioxidant enzymes and polyamines. Chemosphere, 243, 125308. https://doi.org/10.1016/j.chemosphere.2019.125308
  33. Shah, A.A., Ahmed, S., Yasin, N.A. (2020b). 2-Hydroxymelatonin induced nutritional orchestration in Cucumis sativus under cadmium toxicity: modulation of non-enzymatic antioxidants and gene expression. Int. J. Phytoremediat., 22(5), 497‒507. https://doi.org/10.1080/15226514.2019.1683715
  34. Shah, A.A., Yasin, N.A, Ahmed, S. et al. (2021). 4-Hydroxymelatonin alleviates nickel stress, improves physiochemical traits of Solanum melongena: regulation of polyamine metabolism and antioxidative enzyme. Sci. Hortic., 282, 110036. https://doi.org/10.1016/j.scienta.2021.110036
  35. Tan, D.X., Hardeland, R., Manchester, L.C. et al. (2014). Cyclic-3-hydroxymelatonin (C3HOM), a potent antioxidant, scavenges free radicles and suppresses oxidative reactions. Curr. Med. Chem., 21 (13), 1557–1565. http://dx.doi.org/10.2174/0929867321666131129113146
  36. Tan, D.X., Reiter, R.J. (2020). An evolutionary view of melatonin synthesis and metabolism related to its biological func-tions in plants. J. Exp. Bot., 71(16), 4677‒4689. https://doi.org/10.1093/jxb/eraa235
  37. Wang, X.N., Yang, F., Zhang, J.C. et al. (2023). Ectopic expression of MmCYP1A1, a mouse cytochrome P450 gene, positively regulates stress tolerance in apple calli and Arabidopsis. Plant Cell Rep., 42(2), 433‒448. https://doi.org/10.1007/s00299-022-02969-5

Downloads

Download data is not yet available.

Similar Articles

<< < 67 68 69 70 71 72 73 74 75 76 > >> 

You may also start an advanced similarity search for this article.