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Plant Science Today (PST)

Research Articles

Early Access

Comparative analysis of enzymatic and non-enzymatic antioxidant systems in in vivo and in vitro plant parts of Plectranthus amboinicus (Lour.) Spreng.

DOI
https://doi.org/10.14719/pst.13884
Submitted
28 January 2026
Published
28-09-2026
Versions

Abstract

Plectranthus amboinicus (Lour.) Spreng. (Lamiaceae) is a medicinal plant known for its antioxidant properties; however, comparative information on antioxidant systems across different plant parts and growth conditions (in vivo vs in vitro) remains limited. This study quantified major secondary metabolites and evaluated enzymatic and non-enzymatic antioxidant activities in in vivo and in vitro propagated plant parts (leaf, root, stem) as well as callus cultures. Enzymatic antioxidant activities were assessed using catalase (CAT), peroxidase (POX) and superoxide dismutase (SOD) assays, while non-enzymatic antioxidant activity was evaluated using the DPPH assay. In vivo roots showed the highest antioxidant potential (IC50 = 24.56 ± 3.44 µg/mL), followed by in vivo leaves (IC50 = 40.28 ± 6.81 µg/mL), in contrast to ascorbic acid (IC50  = 26.51 ± 3.14 µg/mL) used as standard. The antioxidant enzymes activity ranged from 0.0011 ± 0.0001–0.0031 ± 0.0002 ΔOD min-1 g-1 fresh weight (FW) for CAT, 0.115 ± 0.052–0.525 ± 0.012 ΔOD min-¹ g-¹ FW for POX and 0.0104 ± 0.0002–0.0194 ± 0.0001 units mg-¹ FW for SOD. The results demonstrate that not only leaves but other plant parts (root and stem) and callus also contain higher antioxidant potential Further studies should focus on the isolation, identification and comprehensive characterisation of major bioactive constituents to explore their potential applications in human health.

References

  1. 1. Rao KVM. Introduction. In: Rao KVM, Raghavendra AS, Reddy KJ, editors. Physiology and Molecular Biology of Stress Tolerance in Plants. Dordrecht: Springer Netherlands; 2006. p. 1–14.
  2. 2. Di Meo S, Reed TT, Venditti P, Victor VM. Role of ROS and RNS sources in physiological and pathological conditions. Oxid Med Cell Longev. 2016;2016:1–44. https://doi.org/10.1155/2016/1245049
  3. 3. Altemimi A, Lakhssassi N, Baharlouei A, Watson D, Lightfoot D. Phytochemicals: Extraction, isolation and identification of bioactive compounds from plant extracts. Plants. 2017;6(4):42. https://doi.org/10.3390/plants6040042
  4. 4. Taghvaei M, Jafari SM. Application and stability of natural antioxidants in edible oils in order to substitute synthetic additives. J Food Sci. Technol. 2015;52(3):1272–82. https://doi.org/10.1007/s13197-013-1080-1
  5. 5. Sharma T, Gamit R, Acharya R, Shukla VJ. Quantitative estimation of total tannin, alkaloid, phenolic and flavonoid content of the root, leaf and whole plant of Byttneria herbacea Roxb. AYU. 2023;42:143–7. https://doi.org/10.4103/ayu.AYU_25_19
  6. 6. Afrieyie DK, Ameyaw EO, Acheampong DO, Tuffour I, Opong RA. In vitro and in vivo antioxidant properties of stem extracts of Croton membranaceus. Int J Pharm Sci Res. 2022;13(2):921–9.
  7. 7. Nguyen VT, Nguyen MT, Nguyen NQ, Truc TT. Phytochemical screening, antioxidant activities, total phenolics and flavonoids content of leaves from Persicaria odorata (Polygonaceae). IOP Conf Ser Mater Sci Eng. 2020;991(1):012029.
  8. https://doi.org/10.1088/1757-899X/991/1/012029
  9. 8. Navarrete P, Pizzi A, Pasch H, Rode K, Delmotte L. MALDI-TOF and 13C NMR characterization of maritime pine industrial tannin extract. Ind Crops Prod. 2010;32(2):105–10. https://doi.org/10.1016/j.indcrop.2010.03.010
  10. 9. Lokhande VH, Nikam TD, Patade VY, Ahire ML, Suprasanna P. Effects of optimal and supra-optimal salinity stress on antioxidative defence, osmolytes and in vitro growth responses in Sesuvium portulacastrum L. Plant Cell Tissue Organ Cult. 2011;104(1):41–9. https://doi.org/10.1007/s11240-010-9802-9
  11. 10. Roy D, Mallick B, Samanta D. Augmentation of antioxidative potential of in vitro propagated Mentha piperita L. Indian J Exp Biol. 2020;58:131–7. https://doi.org/10.56042/ijeb.v58i02.65344
  12. 11. Vijyanand S, Thriruvengadam SR, Malathi R. Comparative study on free radical ameliorating potential of stem and the leaf extracts of Plectranthus amboinicus. J Stress Physiol Biochem. 2013;19(1):111–9.
  13. 12. Kumaran A, Karunakaran RJ. Antioxidant and free radical scavenging activity of an aqueous extract of Coleus aromaticus. Food Chem. 2006;97(1):109–14. https://doi.org/10.1016/j.foodchem.2005.03.032
  14. 13. Inoue KH, Hagerman AE. Determination of gallotannin with rhodanine. Anal Biochem. 1988;169(2):363–9. https://doi.org/10.1016/0003-2697(88)90296-5
  15. 14. Makkar HPS. Measurement of total phenolics and tannins using Folin-Ciocalteu method. In: Quantification of tannins in tree and shrub foliage. Dordrecht: Springer; 2003. p. 49–51. https://doi.org/10.1007/978-94-017-0273-7_3
  16. 15. Khatiwora E, Vaishali B, Adsulkulkarni MM, Deshpande NR, Kashalkar RV. Spectroscopic determination of total phenol and flavonoid contents of Ipomoea carnea. Int J ChemTech Res. 2010;2(3):1698–701.
  17. 16. Aebi H. Catalase in vitro. Methods Enzymol. 1984;105:121–6. https://doi.org/10.1016/S0076-6879(84)05016-3
  18. 17. Chance B, Maehly AC. Assays of catalases and peroxidases. Methods Enzymol. 1955;2:764–75.
  19. https://doi.org/10.1016/S0076-6879(55)02300-8
  20. 18. Nasir NA, Roslly NAL, Rosli NM, Razali Z. Phytochemical screening and potential DPPH radical scavenging activity of banana peel extract. In: Changing Sustainable Future ASEAN Sci Technol. 2020. p. 249–58. https://doi.org/10.1007/978-981-15-3434-8_22
  21. 19. Shimamura T, Sumikura Y, Yamazaki T, Tada A, Kashiwagi T, Ishikawa H, et al. Applicability of the DPPH assay for evaluating the antioxidant capacity of food additives: Inter-laboratory evaluation. Anal Sci. 2014;30(7):717–21. https://doi.org/10.2116/analsci.30.717
  22. 20. Mohan B, Ramalingam S, Saravanan R, Dheeba B. In vitro antioxidant and anti-proliferative activity of Plectranthus amboinicus leaves extract on MCF-7 cell line. Der Pharm Lett. 2016;8(12):1–9.
  23. 21. Harborne JB. Phytochemical Methods: A guide to modern techniques of plant analysis. 3rd ed. London: Chapman & Hall; 1997.
  24. 22. Taiz L, Zeiger E, Møller IM, Murphy A. Plant Physiology and Development. Sunderland (MA): Sinauer Associates; 2015.
  25. 23. Dixon RA, Paiva NL. Stress-induced phenylpropanoid metabolism. Plant Cell. 1995;7(7):1085–97. https://doi.org/10.2307/3870059
  26. 24. Rao RS, Ravishankar GA. Plant cell cultures: Chemical factories of secondary metabolites. Biotechnol Adv. 2002;20(2):101–53. https://doi.org/10.1016/S0734-9750(02)00007-1
  27. 25. Matkowski A. Plant in vitro culture for the production of antioxidants: A review. Biotechnol Adv. 2008;26(6):548–60. https://doi.org/10.1016/j.biotechadv.2008.07.001
  28. 26. Sreekala V, Vijaykishan B, Rajasekar SS, Rajesh R. Phytochemical analysis and in vitro DPPH assay of ethanolic extract of Mucuna pruriens seed. Int J Curr Pharm Res. 2020;12:693–6. https://doi.org/10.22159/ijcpr.2020v12i6.40300
  29. 27. Bourgaud F, Gravot A, Milesi S, Gontier E. Production of plant secondary metabolites: A historical perspective. Plant Sci. 2001;161(5):839–51. https://doi.org/10.1016/S0168-9452(01)00490-3
  30. 28. Murthy HN, Lee EJ, Paek KY. Production of secondary metabolites from cell and organ cultures: Strategies and approaches for biomass improvement and metabolite accumulation. Plant Cell Tissue Organ Cult. 2014;118(1):1–16.
  31. https://doi.org/10.1007/s11240-014-0467-7
  32. 29. Nasirkandi AM, Alirezalu A, Alipour A, Amato J. Screening of 20 species from Lamiaceae family based on phytochemical analysis, antioxidant activity and HPLC profiling. Sci Rep. 2023;13:16987. https://doi.org/10.1038/s41598-023-44337-7
  33. 30. Magulska BU, Wesolowski M. Antioxidant activity of medicinal herbs and spices from plants of the Lamiaceae, Apiaceae and Asteraceae families: Chemometric interpretation of the data. Antioxidants. 2023;12:2039. https://doi.org/10.3390/antiox12122039
  34. 31. Winkel-Shirley B. Biosynthesis of flavonoids and effects of stress. Curr Opin Plant Biol. 2002;5(3):218–23. https://doi.org/10.1016/S1369-5266(02)00256-X
  35. 32. Agati G, Azzarello E, Pollastri S, Tattini M. Flavonoids as antioxidants in plants: Location and functional significance. Plant Sci. 2012;196:67–76. https://doi.org/10.1016/j.plantsci.2012.07.014
  36. 33. Verpoorte R, Contin A, Memelink J. Biotechnology for the production of plant secondary metabolites. Phytochem Rev. 2002;1(1):13–25. https://doi.org/10.1023/A:1015871916833
  37. 34. Akula R, Ravishankar GA. Influence of abiotic stress signals on secondary metabolites in plants. Plant Signal Behav. 2011;6(11):1720–31. https://doi.org/10.4161/psb.6.11.17613
  38. 35. Rai MK, Kalia RK, Singh R, Gangola MP, Dhawan AK. Developing stress tolerant plants through in vitro selection: An overview of the recent progress. Environ Exp Bot. 2011;71(1):89–98. https://doi.org/10.1016/j.envexpbot.2010.10.021
  39. 36. Nathawat RS. In vitro Propagation and Phytochemical Studies on Fracture Healing Plants: Cissus quadrangularis L. and Lepidium sativum L. [PhD thesis]. Rajasthan: University of Rajasthan; 2013.
  40. 37. Gunes T. Peroxidase and IAA-oxidase activities during rooting in cuttings of three poplar species. Turk J Bot. 2000;24:97–101.
  41. 38. 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
  42. 39. Mishra A, Singh B, Kumar C. Comparative antioxidant enzyme activities in leaves, stems and roots of selected medicinal and aromatic plants. J Med Plant Res. 2013;7(18):1320–8.
  43. 40. Rout JR, Sahoo SL. In vitro propagation and antioxidant enzyme activities of Elephantopus scaber L. Aspac J Mol Biol Biotechnol. 2013;21(2):59–66.
  44. 41. Anita A, Abisha MJ, Jomol JC, Singh RH, Mettel SS. Phytochemical screening and antioxidant activity of Plectranthus amboinicus. J Emerg Technol Innov Res. 2024;7:777–83.
  45. 42. Joshi R. In vitro and in vivo biochemical studies in Merremia species [PhD thesis]. Rajasthan: University of Rajasthan; 2018.
  46. 43. Bhandari J. Study of phytochemical, anti-microbial, antioxidant and anti-cancer properties of Allium wallichii. BMC Complement Altern Med. 2017;17:102. https://doi.org/10.1186/s12906-017-1622-6
  47. 44. 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
  48. 45. Phongpaichit S, Nikom J, Rungjindamai N, Sakayaroj J, Hutadilok-Towatana N, Rukachaisirikul V, et al. Biological activities of extracts from endophytic fungi isolated from Garcinia plants. FEMS Immunol Med Microbiol. 2007;51(3):517–25.
  49. https://doi.org/10.1111/j.1574-695X.2007.00331.x
  50. 46. Lisiewska Z, Kmiecik W, Korus A. Content of vitamin C, carotenoids, chlorophylls and polyphenols in green parts of dill (Anethum graveolens L.) depending on plant height. J Food Compos Anal. 2006;19(2–3):134–40. https://doi.org/10.1016/j.jfca.2005.04.009

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