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Plant Science Today (PST)

Research Articles

Early Access

Therapeutic potential and metabolite profiling of fungal endophytes from Withania somnifera (L.) Dunal

DOI
https://doi.org/10.14719/pst.11888
Submitted
21 September 2025
Published
17-10-2026

Abstract

Endophytic fungi associated with Withania somnifera (L.) Dunal represent a promising source of bioactive compounds, some of which may be identical to or distinct from those produced by the host plant. However, their metabolic diversity and therapeutic potential, influenced by geography and environment, remain underexplored. The crude extracts of the fungal endophytic isolates were tested for their antioxidant activity, antidiabetic potential and anti-inflammatory efficacy by established biochemical assays. When compared with other endophytic isolates, namely W2P, W1R, W2S, W1L and W2LA, W2LB consistently showed the highest activity in phosphomolybdenum and ferric reducing antioxidant power (FRAP) assays, suggesting substantial antioxidant potential. Endophytic isolate W1L, on the other hand, demonstrated the highest 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and high anti-inflammatory activity. Internal transcribed spacer (ITS) rDNA sequencing identified two potent fungal endophytic isolates, W1L and W2LB as Daldinia korfii and Diaporthe perseae. The extract’s liquid chromatography-mass spectrometry (LC-MS) and gas chromatography-mass spectrometry (GC-MS) profiling revealed a broad range of phenolics, terpenoids, alkaloids, alkanes, fatty acids and other pharmaceutically significant compounds. This study adds to the diversity of fungal endophytes associated with W. somnifera by identifying less frequently found species, including D. perseae, D. korfii and indicates that the endophytes of W. somnifera enhance the phytochemical diversity of their host, contributing to anti-inflammatory, antidiabetic and antioxidant activities. Hence, the experiment supports further purification and analysis for drug development.

References

  1. 1. Aamir M, Rai KK, Zehra A, Kumar S, Yadav M, Shukla V, et al. Fungal endophytes: Classification, diversity, ecological role and their relevance in sustainable agriculture. In: Kumar A, Singh VK, editors. Microbial endophytes: Prospects for sustainable agriculture. Woodhead Publishing Series in Food Science, Technology and Nutrition. Cambridge: Woodhead Publishing; 2020. p. 291–323. https://doi.org/10.1016/B978-0-12-818734-0.00012-7
  2. 2. Sethi M, Kaur C, Hagroo RP, Singh MP. Endophyte mediated plant health via phytohormones and biomolecules. In: Solanki MK, Yadav MK, Singh BP, Gupta VK, editors. Microbial endophytes and plant growth: Beneficial interactions and applications. Cambridge: Academic Press; 2023. p. 151–66. https://doi.org/10.1016/B978-0-323-90620-3.00017-9
  3. 3. Debbab A, Aly AH, Proksch P. Bioactive secondary metabolites from endophytes and associated marine derived fungi. Fungal Divers. 2011;49:1–12. https://doi.org/10.1007/s13225-011-0114-0
  4. 4. Bharti VK, Malik JK, Gupta RC. Ashwagandha: Multiple health benefits. In: Gupta RC, Lall R, Srivastava A, editors. Nutraceuticals: Efficacy, safety and toxicity. 2nd ed. Cambridge: Academic Press; 2021. p. 865–80. https://doi.org/10.1016/B978-0-12-821038-3.00050-1
  5. 5. Singh P, Guleri R, Pati PK. In vitro propagation of Withania somnifera (L.) Dunal. In: Jain SM, editor. Protocols for in vitro cultures and secondary metabolite analysis of aromatic and medicinal plants. 2nd ed. New York, NY: Humana Press; 2016. p. 201–13. https://doi.org/10.1007/978-1-4939-3332-7_14
  6. 6. Kaul S, Ahmed M, Sharma T, Dhar MK. Unlocking the myriad benefits of endophytes: An overview. In: Kharwar RN, Upadhyay RS, Dubey NK, Raghuwanshi R, editors. Microbial diversity and biotechnology in food security. New Delhi: Springer India; 2014. p. 41–57. https://doi.org/10.1007/978-81-322-1801-2_4
  7. 7. Kushwaha RK, Singh S, Pandey SS, Kalra A, Babu CSV. Fungal endophytes attune withanolide biosynthesis in Withania somnifera, prime to enhanced withanolide A content in leaves and roots. World J Microbiol Biotechnol. 2019;35(2):20. https://doi.org/10.1007/s11274-019-2593-1
  8. 8. Muthukrishnan S, Prakathi P, Sivakumar T, Thiruvengadam M, Jayaprakash B, Baskar V, et al. Bioactive components and health potential of endophytic micro-fungal diversity in medicinal plants. Antibiotics. 2022;11(11):1533. https://doi.org/10.3390/antibiotics11111533
  9. 9. Charria-Girón E, Espinosa MC, Zapata-Montoya A, Méndez MJ, Caicedo JP, Dávalos AF, et al. Evaluation of the antibacterial activity of crude extracts obtained from cultivation of native endophytic fungi belonging to a tropical montane rainforest in Colombia. Front Microbiol. 2021;12:716523. https://doi.org/10.3389/fmicb.2021.716523
  10. 10. Dwivedi D, Yadav B, Sengar N. Extraction and isolation of withaferin A (steroidal lactone) from Withania somnifera leaf and its TLC and HPLC analysis. Orient J Chem. 2017;33(3):1550–4. https://doi.org/10.13005/ojc/330360
  11. 11. Ayele DT, Akele ML, Melese AT. Analysis of total phenolic contents, flavonoids, antioxidant and antibacterial activities of Croton macrostachyus root extracts. BMC Chem. 2022;16:30. https://doi.org/10.1186/s13065-022-00822-0
  12. 12. Gurgel RS, de Melo PDÍ, Garcia AVF, de Souza ATF, da Silva TM, de Andrade CP, et al. Antimicrobial and antioxidant activities of endophytic fungi associated with Arrabidaea chica (Bignoniaceae). J Fungi. 2023;9(8):864. https://doi.org/10.3390/jof9080864
  13. 13. Bhat PR, Trushti GS. Therapeutic importance of extracts from leaf and bark of Gymnacranthera farquhariana. J Nat Remedies. 2024;24(9):1975–85. https://doi.org/10.18311/jnr/2024/41899
  14. 14. Bhatti MZ, Ali A, Ahmad A, Saeed A, Malik SA. Antioxidant and phytochemical analysis of Ranunculus arvensis L. extracts. BMC Res Notes. 2015;8:279. https://doi.org/10.1186/s13104-015-1228-3
  15. 15. Moharram AM, Zohri AA, Omar HM, El-Ghani OAA. In vitro assessment of antimicrobial and anti-inflammatory potential of endophytic fungal metabolites extracts. Eur J Biol Res. 2017;7(3):234–44.
  16. 16. Samapti MMS, Afroz F, Rony SR, Sharmin S, Moni F, Akhter S, et al. Isolation and identification of endophytic fungi from Syzygium cumini Linn and investigation of their pharmacological activities. Sci World J. 2022;2022:9529665. https://doi.org/10.1155/2022/9529665
  17. 17. Elfita, Oktiansyah R, Mardiyanto, Widjajanti H, Setiawan A. Antibacterial and antioxidant activity of endophytic fungi isolated from Peronema canescens leaves. Biodiversitas. 2022;23(9):4783–92. https://doi.org/10.13057/biodiv/d230946
  18. 18. Zhu ZJ, Schultz AW, Wang J, Johnson CH, Yannone SM, Patti GJ, et al. Liquid chromatography quadrupole time-of-flight mass spectrometry characterization of metabolites guided by the METLIN database. Nat Protoc. 2013;8:451–60. https://doi.org/10.1038/nprot.2013.004
  19. 19. Chutulo EC, Chalannavar RK. Daldinia eschscholtzii: An endophytic fungus isolated from Psidium guajava as an alternative source of bioactive secondary metabolites. Asian J Mycol. 2020;3(1):376–98. https://doi.org/10.5943/ajom/3/1/11
  20. 20. Devarajan A, Rao BV, Francis T. Natural and synthetic sources of phenolic antioxidants. In: Upaganlawar AB, Dhote VV, Raja MKMM, editors. Health benefits of phenolic antioxidants. New York: Nova Science Publishers; 2022. p. 1–46.
  21. 21. Jamal HAA, Husaini A, Sing NN, Roslan HA, Zulkharnain A, Akinkunmi WA. Characterization of bioactive compounds produced by endophytic fungi isolated from Gynura procumbens (Sambung Nyawa). Braz J Microbiol. 2022;53:1857–70. https://doi.org/10.1007/s42770-022-00827-w
  22. 22. Magagula M, Motaung TE, Mbita Z, Dithebe K. Investigating the antimicrobial and anticancer potential of culturable fungal endophytes isolated from the stems of Kirkia acuminata Oliv. BMC Microbiol. 2025;25:343. https://doi.org/10.1186/s12866-025-03964-y
  23. 23. Dutta R, Ghosal S, Roy S, Bhattacharyya DK, Bhowal J. Phytochemical screening and antioxidative property evaluation of lipid-producing fungi. Arch Microbiol. 2023;205:65. https://doi.org/10.1007/s00203-022-03399-3
  24. 24. Madki MA, Manjoor AS, Powar PV, Patil KS. Isolation and biological activity of endophytic fungi from Withania somnifera. Int J Pharm Sci. 2010;2(3):848–58.
  25. 25. Niaz SI, Khan D, Naz R, Safdar K, Ul Abidin SZ, Khan IU, et al. Antimicrobial and antioxidant chlorinated azaphilones from mangrove Diaporthe perseae sp. isolated from the stem of Chinese mangrove Pongamia pinnata. J Asian Nat Prod Res. 2021;23(11):1077–84. https://doi.org/10.1080/10286020.2020.1835872
  26. 26. Tan WN, Nagarajan K, Lim V, Azizi J, Khaw KY, Tong WY, et al. Metabolomics analysis and antioxidant potential of endophytic Diaporthe fraxini ED2 grown in different culture media. J Fungi. 2022;8(5):519. https://doi.org/10.3390/jof8050519
  27. 27. Nsimba RY, Kikuzaki H, Konishi Y. Antioxidant activity of various extracts and fractions of Chenopodium quinoa and Amaranthus spp. seeds. Food Chem. 2008;106(2):760–6. https://doi.org/10.1016/j.foodchem.2007.06.004
  28. 28. Pavithra N, Satish L, Suneel KA, Venkatarathanamma V, Pushpalatha H, Bhanuprakash RG, et al. In vitro studies on α-amylase, α-glucosidase and aldose reductase inhibitors found in endophytic fungi isolated from Ocimum sanctum. Curr Enzym Inhib. 2014;10(2):129–36. https://doi.org/10.2174/1573408011666141205233252
  29. 29. Singh B, Kaur A. Antidiabetic potential of a peptide isolated from an endophytic Aspergillus awamori. J Appl Microbiol.2016;120(2):301–11. https://doi.org/10.1111/jam.12998
  30. 30. Khan R, Naqvi STQ, Fatima N, Muhammad SA. Study of antidiabetic activities of endophytic fungi isolated from plants. Pure Appl Biol. 2019;8(2):1287–95. https://doi.org/10.19045/bspab.2019.80071
  31. 31. Sharma G, Agarwal S, Bhardwaj R, Barvkar VT, Godbole RC, Kumar V, et al. Pharmacological potential of Curcuma longa endophytes. Microbe. 2024;5:100187. https://doi.org/10.1016/j.microb.2024.100187
  32. 32. Bhadra F, Vasundhara M. Anti-inflammatory potential of aconitine produced by endophytic fungus Acremonium alternatum. World J Microbiol Biotechnol. 2024;40:274. https://doi.org/10.1007/s11274-024-04083-y
  33. 33. Karthik TD, Krishna V, Sourabh GBU, Raagavalli K, Syeda AS. Evaluation of in vitro antioxidant and anti-inflammatory potentials of phytochemicals identified from leaf extracts of Hibiscus lobatus Kuntze. J Chem Health Risks. 2025;15(3):824–37.
  34. 34. Fu Y, Wang Q, Tang Z, Liu G, Guan G, Lyu J. Cordycepin ameliorates high fat diet-induced obesity by modulating endogenous metabolism and gut microbiota dysbiosis. Nutrients.2024;16(17):2859. https://doi.org/10.3390/nu16172859
  35. 35. Polis BMD, Srikanth KD, Gurevich V, Gil-Henn H, Samson AO. L-Norvaline, a new therapeutic agent against Alzheimer's disease. Neural Regen Res. 2019;14(9):1562–72. https://doi.org/10.4103/1673-5374.255980
  36. 36. Gilinsky MA, Polityko YK, Markel AL, Latysheva TV, Samson AO, Polis B, et al. Norvaline reduces blood pressure and induces diuresis in rats with inherited stress-induced arterial hypertension. Biomed Res Int. 2020;2020:4935386. https://doi.org/10.1155/2020/4935386
  37. 37. Pastuszak I, Molyneux RJ, James LF, Elbein AD. Lentiginosine, a dihydroxyindolizidine alkaloid that inhibits amyloglucosidase. Biochemistry. 1990;29(7):1886–91. https://doi.org/10.1021/bi00459a032
  38. 38. Ching-Wen C, Yun-Chieh C, Yu-Chin L, Wen-Huang P. p-Hydroxyacetophenone suppresses nuclear factor-κB-related inflammation in nociceptive and inflammatory animal models. J Nat Med. 2017;71:422–32.
  39. https://doi.org/10.1007/s11418-017-1074-9
  40. 39. Liu Y, Li C, Wu H, Xie X, Sun, Dai M. Paeonol attenuated inflammatory response of endothelial cells via stimulating monocytes-derived exosomal microRNA-223. Front Pharmacol. 2018;9:1105. https://doi.org/10.3389/fphar.2018.01105
  41. 40. Agudelo LZ, Ferreira DMS, Cervenka I, Bryzgalova G, Dadvar S, Jannig PR, et al. Kynurenic acid and Gpr35 regulate adipose tissue energy homeostasis and inflammation. Cell Metab. 2018;27(2):378–392.e5. https://doi.org/10.1016/j.cmet.2018.01.004
  42. 41. Calado M, Santos D, Susano P, Pinteus S, Martins A, Silva J, et al. Endophytic fungi associated with seaweeds as potential producers of antimicrobial compounds. Mar Biotechnol. 2026;28. https://doi.org/10.1007/s10126-026-10626-1
  43. 42. Nagarajan K, Ibrahim B, Bawadikji A, Khaw KY, Tong WY, Leong CR, et al. Characterization of metabolites in an endophytic fungus Diaporthe fraxini via NMR-based metabolomics and cholinesterase inhibitory activity. Appl Biochem Microbiol. 2023;59:316–22. https://doi.org/10.1134/S0003683823030134
  44. 43. de Arruda Silva M, da Silva LPF, dos Santos ACR, Rocha ACM, dos Anjos Almudi EM, Bellete BS, et al. Metabolomics of Trichoderma spirale by mass spectrometry data dereplication. Chem Nat Compd. 2025;61:881–7. https://doi.org/10.1007/s10600-025-04780-6
  45. 44. Karila T, Tervahartiala T, Cohen B, Sorsa T. An anti-inflammatory agent, 2-hydroxyisocaproic acid, prevents del-1 fragmentation in vitro. Med Res Arch. 2024;12(7). https://doi.org/10.18103/mra.v12i7.5466
  46. 45. Katkeviciute E, Bircher A, Sanchez R, Gademann K, Scharl M, Montalban-Arques A, et al. Bacteria-derived 3-hydroxydodecanoic acid induces a potent anti-tumor immune response via the GPR84 receptor. Cell Rep. 2025;44(3):115357. https://doi.org/10.1016/j.celrep.2025.115357
  47. 46. Sobrinho ACN, Fontenelle RODS, Souza EBD, Morais SMD. Antifungal and antioxidant effect of the lachnophyllum ester, isolated from the essential oil of Baccharis trinervis (Lam.) Pers., against dermatophytes fungi. Rev Bras Saude Prod Anim. 2021;22:1–11. https://doi.org/10.1590/s1519-99402122542021
  48. 47. Rashad YM, Abdalla SA, Shehata AS. Aspergillus flavus YRB2 from Thymelaea hirsuta (L.) Endl., a non-aflatoxigenic endophyte with ability to overexpress defense-related genes against Fusarium root rot of maize. BMC Microbiol. 2022;22(1):229. https://doi.org/10.1186/s12866-022-02651-6
  49. 48. Balachandran A, Choi SB, Beata MM, Małgorzata J, Froemming GRA, Lavilla CA Jr, et al. Antioxidant, wound healing potential and in silico assessment of naringin, eicosane and octacosane. Molecules. 2023;28(3):1043. https://doi.org/10.3390/molecules28031043
  50. 49. Okechukwu PN. Evaluation of anti-inflammatory, analgesic, antipyretic effect of eicosane, pentadecane, octacosane, and heneicosane. Asian J Pharm Clin Res. 2020;13(4). https://doi.org/10.22159/ajpcr.2020.v13i4.36196
  51. 50. Jamal Uddin S, Grice D, Tiralongo E. Evaluation of cytotoxic activity of patriscabratine, tetracosane and various flavonoids isolated from the Bangladeshi medicinal plant Acrostichum aureum. Pharm Biol. 2012;50(10):1276–80. https://doi.org/10.3109/13880209.2012.673628
  52. 51. Agrahari S, Singh SP, Singh BN. Bioactive metabolites and antidiabetic activity of Cannabis sativa-derived endophytic fungi. Arch Microbiol. 2026;28. https://doi.org/10.21203/rs.3.rs-7549570/v1
  53. 52. Owolabi MS, Lawal OA, Ogunwande IA, Hauser RM, Setzer WN. Chemical composition of the leaf essential oil of Terminalia catappa L. growing in southwestern Nigeria. Am J Essent Oil Nat Prod. 2013;1(1):51–4.

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