Bioprospection unveils the bioactive potential of Colletotrichum taiwanense BPSRJ3, an endophytic fungus of an ethnomedicinal orchid, Vanda cristata Wall. Ex Lindl.

Abstract

Endophytic fungi associated with medicinal plants are reservoirs of compounds with therapeutic properties. The present study focused on the isolation and identification of endophytic fungi from an ethnomedicinal orchid, Vanda cristata Wall. ex Lindl., and the assessment of their bioactive potential through screening their phytochemical content, antioxidant property, antimicrobial activity, and cytotoxicity, followed by metabolite profiling of the promising isolate extract. Colletotrichum taiwanense BPSRJ3 extract showed the highest total phenolic and flavonoid content. Although all isolate extracts showed broad spectrum antimicrobial activity, C. taiwanense BPSRJ3 extract showed the widest zone of inhibition and lowest MIC against the test pathogens. It also exhibited cytotoxicity against MCF-7 and A549 human cancer cell lines while having no discernible cytotoxic impact on the HEK-293 human normal cell line. Metabolites of C. taiwanense BPSRJ3 extract were characterized using FT-IR spectroscopy and GC–MS analysis, providing insights into the biologically active compounds contributing to its bioactivity. It is the first study to report C. taiwanense isolated from V. cristata as having bioactive potential. The findings of this study provide opportunities for more investigation into using the advantages of this fungal endophyte in the pharmaceutical sector.

Graphical abstract

Keywords

Vanda cristata / Antioxidant / Antimicrobial / Cytotoxicity / Collectotrichum taiwanense

References

[1] Younessi-HamzekhanluM, OzturkM, AltayV, NojadehMS, AlakbarliF. Ethnopharmacological study of medicinal plants from Khoy city of West Azerbaijan-Iran. Indian J Tradit Knowl, 2020, 19: 251-267

[2] AbeyrathneEDNS, NamK, HuangX, AhnDU. Plant- and animal-based antioxidants’ structure, efficacy, mechanisms, and applications: a review. Antioxidants, 2022, 11: 1025.

[3] PancuDF, ScurtuA, MacasoiIG, MartiD, MiocM, SoicaC, et al. . Antibiotics: conventional therapy and natural compounds with antibacterial activity-A pharmaco-toxicological screening. Antibiotics, 2021, 10: 401.

[4] McAlpine JB, Pazoles C, Stafford A. Phytera’s strategy for the discovery of novel anti-infective agents from plant cell cultures. Bioassay methods in natural product research and drug development. Dordrecht: Springer; 1999. pp. 159–66. https://doi.org/10.1007/978-94-011-4810-8_13.

[5] StoneJK, CharlesWB, JamesF, WhiteJWhiteJ. An overview of endophytic microbes: endophytism defined. Microbial endophytes, 2000New YorkMarcel Dekker Inc.17-44

[6] BaconCW, WhiteJMicrobial endophytes, 20001New YorkMarcel Dekker.

[7] StrobelGA. Endophytes as sources of bioactive products. Microbes Infect, 2003, 5: 535-544.

[8] ManganyiMC, AtebaCN. Untapped potentials of endophytic fungi: a review of novel bioactive compounds with biological applications. Microorganisms, 2020, 8: 1934.

[9] UzmaF, MohanCD, HashemA, KonappaNM, RangappaS, KamathPV, et al. . Endophytic fungi-alternative sources of cytotoxic compounds: a review. Front Pharmacol, 2018, 9: 309.

[10] TanRX, ZouWX. Endophytes: a rich source of functional metabolites (1987 to 2000). Nat Prod Rep, 2001, 18: 448-459.

[11] MedhiRP, ChakrabartiS. Traditional Knowledge of NE people on conservation of wild orchids. Indian J Tradit Knowl, 2009, 8: 11-16

[12] PathakP, KumariA, ChandlerBD, ZettlerLW. In vitro propagation and phytochemical analysis of Vanda cristata Wall. ex Lindl.: an endangered medicinal orchid of biopharmaceutical importance. S Afr J Bot, 2023, 153: 109-123.

[13] ChandMB, PaudelMR, PantB. The antioxidant activity of selected wild orchids of Nepal. J Coast Life Med, 2016, 4: 731-736.

[14] JoshiPR, PaudelMR, ChandMB, PradhanS, PantKK, JoshiGP, et al. . Cytotoxic effect of selected wild orchids on two different human cancer cell lines. Heliyon, 2020, 6: e03991.

[15] PaulS, BhagobatyRK, NihalaniMC, JoshiSR. Characterization of oleaginous endophytic fungi of biodiesel plants as potential biofuel minifactories. Biomass Bioenergy, 2020, 142: 105750.

[16] TamuraK, StecherG, KumarS. MEGA11: molecular evolutionary genetics analysis version 11. Mol Biol Evol, 2021, 38: 3022-3027.

[17] KhalilD, El-ZayatSA, El-SayedMA. Phytochemical screening and antioxidant potential of endophytic fungi isolated from Hibiscus sabdariffa. J Appl Biotechnol Rep, 2020, 7: 116-124

[18] DekaD, JhaDK. Bioactivity assessment of endophytic fungi associated with Citrus macroptera Montr.: an endangered ethnomedicinal plant used in folk medicines in North-East India. Indian Phytopathol, 2020, 73: 21-33.

[19] PourmoradF, HosseinimehrSJ, ShahabimajdN. Antioxidant activity, phenol and flavonoid contents of some selected Iranian medicinal plants. Afr J Biotechnol, 2006, 5: 1142-1145

[20] Chang CC, Yang MH, Wen HM, Chern JC. Estimation of total flavonoid content in propolis by two complementary colometric methods. J Food Drug Anal. 2020;10. https://doi.org/10.38212/2224-6614.2748.

[21] GhareebMA, HamdiSA, FolMF, IbrahimAM. Chemical characterization, antibacterial, antibiofilm, and antioxidant activities of the methanolic extract of Paratapes undulatus clams (Born, 1778). J Appl Pharm Sci, 2022.

[22] Chandana L, Bhikshapathi D. Free radical scavenging activity of Pleurotus ostreatus against CCL₄-induced hepatic damage in Wister rats. Int J Pharm Pharm Sci 2023:17–22. https://doi.org/10.22159/ijpps.2023v15i12.49478.

[23] ChutuloEC, ChalannavarRK. Daldinia eschscholtzii: an endophytic fungus isolated from Psidium guajava as an alternative source of bioactive secondary metabolites. Asian J Mycol, 2020, 3: 376-398.

[24] ChutuloE, ChalannavarRK, KumarPP. Chemical compositions, cytotoxicity and antioxidant activity of the endophytic fungus Fusarium napiforme isolated from Psidium guajava. Stud Fungi, 2020, 5: 332-352.

[25] PervezMR, MusaddiqM, ThakarePV, KumarA. Characterization of bioactive compound isolated from Myrothecium spp. with UV, FTIR and HPLC analysis. Indian J Pharm Biol Res, 2015, 3: 1-5.

[26] GribbleGThe alkaloids: chemistry and biology, 1998GA CordellAcademic Press, Inc.51

[27] DeBT, PietersL, DeelstraH, VlietinckA. Condensed vegetable tannins: biodiversity in structure and biological activities. Biochem Syst Ecol, 1999, 27: 445-459.

[28] McCaskillD, CroteauR. Some caveats for bioengineering terpenoid metabolism in plants. Trends Biotechnol, 1998, 16: 349-355.

[29] LafayS, Gil-IzquierdoA. Bioavailability of phenolic acids. Phytochem Rev, 2008, 7: 301-311.

[30] TapasA, SakarkarD, KakdeR. Flavonoids as nutraceuticals: a review. Trop J Pharm Res, 2008, 7: 1089-1099.

[31] Ioset JR, Brun R, Wenzler T, Kaiser M, Yardley V. Drug screening for kinetoplastids diseases: a training manual for screening in neglected diseases. 2009.

[32] KusariS, PandeySP, SpitellerM. Untapped mutualistic paradigms linking host plant and endophytic fungal production of similar bioactive secondary metabolites. Phytochemistry, 2013, 91: 81-87.

[33] SharmaD, PramanikA, AgrawalPK. Evaluation of bioactive secondary metabolites from endophytic fungus Pestalotiopsis neglecta BAB-5510 isolated from leaves of Cupressus torulosa D.Don. 3 Biotech, 2016, 6: 210.

[34] Hernández-RamírezF, DamonA, Fernández PavíaSP, Guillén-NavarroK, Iracheta-DonjuanL, ZarzaE, et al. . Community richness and diversity of endophytic fungi associated with the orchid Guarianthe skinneri infested with “Black Blotch” in the Soconusco Region, Chiapas, Mexico. Diversity, 2023, 15: 807.

[35] SohrabiM, SamsampourD, BagheriA. Molecular identification of fungal endophytes of medicinal plant Citrullus colocynthis (L.) Schrad as a medicinal plant: role of tissue type and sampling location on the diversity. Mol Biotechnol, 2024, 66: 424-431.

[36] Riga R, Wardatillah R, Suryani O, Ryplida B, Suryelita S, Azhar M, et al. Endophytic fungus from Gynura japonica: phytochemical screening, biological activities, and characterisation of its bioactive compound. Nat Prod Res. 2024;1–9. https://doi.org/10.1080/14786419.2024.2332947.

[37] NongthombamKS, MutumSS, PandeyRR. Secondary metabolite profiling of the endophytic fungus Trichoderma longibrachiatum L2D2 isolated from Anaphalis contorta. Annu Res Rev Biol, 2024, 39: 1-15.

[38] YadavM, YadavA, YadavJP. In vitro antioxidant activity and total phenolic content of endophytic fungi isolated from Eugenia jambolana Lam. Asian Pac J Trop Med, 2014, 7: S256-S261.

[39] GautamVS, SinghA, KumariP, NishadJH, KumarJ, YadavM, et al. . Phenolic and flavonoid contents and antioxidant activity of an endophytic fungus Nigrospora sphaerica (EHL2), inhabiting the medicinal plant Euphorbia hirta (dudhi) L. Arch Microbiol, 2022, 204: 140.

[40] BoraP, DeviNN. Exploration of the chemical constituents and its antioxidant, antibacterial activities of endophytic fungi isolated from the medicinal plant Dillenia indica. Arch Microbiol, 2023, 205: 67.

[41] BhagobatyRK, JoshiSR, LekarskaM. Antimicrobial and antioxidant activity of endophytic fungi isolated from ethnomedicinal plants of the “Sacred forests” of Meghalaya. India Med Mycol, 2012, 19: 5-11

[42] WenJ, OkyereSK, WangJ, HuangR, WangY, LiuL, et al. . Endophytic fungi isolated from Ageratina adenophora exhibits potential antimicrobial activity against multidrug-resistant Staphylococcus aureus. Plants, 2023, 12: 650.

[43] JaneTNO, ChukwubuikemCO, OgechiOA, EmmanuelCO, ChigozieCE, OnyekaCO, et al. . Antimicrobial activity of secondary metabolites produced by Colletotrichum species, an endophytic fungus on Vernonia amygdalina Del (fam. Asteraceae). World J Biol Pharm Health Sci, 2023, 14: 31-42.

[44] KumariM, TaritlaS, SharmaA, JayabaskaranC. Antiproliferative and antioxidative bioactive compounds in extracts of marine-derived endophytic fungus Talaromyces purpureogenus. Front Microbiol, 2018, 9: 1777.

[45] PedraNS, CanutoKM, de Queiroz SouzaAS, RibeiroPRV, BonaNP, Ramos-SobrinhoR, et al. . Endophytic fungus of Achyrocline satureioides: molecular identification, chemical characterization, and cytotoxic evaluation of its metabolites in human melanoma cell line. Appl Biochem Biotechnol, 2023, 195: 4011-4035.

[46] SharafMH, AbdelazizAM, KalabaMH, RadwanAA, HashemAH. Antimicrobial, antioxidant, cytotoxic activities and phytochemical analysis of fungal endophytes isolated from Ocimum Basilicum. Appl Biochem Biotechnol, 2022, 194: 1271-1289.

[47] DanagoudarA, JoshiC, RaviS, Rohit KumarH, RameshB. Antioxidant and cytotoxic potential of endophytic fungi isolated from medicinal plant Tragia involucrata L. Pharmacognosy Res, 2018, 10: 188.

[48] Kukula-Koch WA, Widelski J. Alkaloids. Pharmacognosy. Elsevier; 2017. pp. 163–98. https://doi.org/10.1016/B978-0-12-802104-0.00009-3.

[49] Ludwiczuk A, Skalicka-Woźniak K, Georgiev MI. Terpenoids. Pharmacognosy. Elsevier; 2017, pp. 233–66. https://doi.org/10.1016/B978-0-12-802104-0.00011-1.

[50] MiaoF, YangR, ChenD-D, WangY, QinB-F, YangX-J, et al. . Isolation, identification and antimicrobial activities of two secondary metabolites of Talaromyces verruculosus. Molecules, 2012, 17: 14091-14098.

[51] MilaevaER, GerasimovaOA, JingweiZ, ShpakovskyDB, SyrbuSA, SemeykinAS, et al. . Synthesis and antioxidative activity of metalloporphyrins bearing 2,6-di-tert-butylphenol pendants. J Inorg Biochem, 2008, 102: 1348-1358.

[52] AntonenkoTA, ShpakovskyDB, VorobyovMA, GrachevaYA, KharitonashviliEV, DubovaLG, et al. . Antioxidative vs cytotoxic activities of organotin complexes bearing 2,6-di-tert-butylphenol moieties. Appl Organomet Chem, 2018, 32: e4381.

[53] BoudjadaA, TouilA, BensouiciC, BendifH, RhouatiS. Phenanthrene and dihydrophenanthrene derivatives from Dioscorea communis with anticholinesterase, and antioxidant activities. Nat Prod Res, 2019, 33: 3278-3282.

[54] NischithaR, VasanthkumariMM, KumaraswamyBE, ShivannaMB. Antimicrobial and antioxidant activities and chemical profiling of Curvularia tsudae endophytic in Cynodon dactylon (L.) Pers. 3 Biotech, 2020, 10: 300.

[55] VermaA, RaiN, GuptaP, SinghS, TiwariH, ChauhanSB, et al. . Exploration of in vitro cytotoxic and in ovo antiangiogenic activity of ethyl acetate extract of Penicillium oxalicum. Environ Toxicol, 2023, 38: 2509-2523.

[56] MaitraS, DeA, DasB, RoySN, ChakrabortyR, SamantaA, et al. . Seasonal variation of phyto-constituents of tea leaves afects antiproliferative potential. J Am Coll Nutr, 2019, 38: 415-423.

[57] NishadJH, SinghA, GautamVS, KumariP, KumarJ, YadavM, et al. . Bioactive potential evaluation and purification of compounds from an endophytic fungus Diaporthe longicolla, a resident of Saraca asoca (Roxb.) Willd. Arch Microbiol, 2021, 203: 4179-4188.

[58] FarhatH, UroojF, SohailN, UllahS, AamerM. Evaluation of antimicrobial potential of endophytic fungi and GC-MS metabolic profiling of Cephalosporium sp., and Fusarium moniliforme. J Exp Pathol, 2023, 4: 16-23.

[59] ChohanTA, SarfrazM, RehmanK, MuhammadT, GhoriMU, KhanKM, et al. . Phytochemical profiling, antioxidant and antiproliferation potential of Euphorbia milii var.: experimental analysis and in-silico validation. Saudi J Biol Sci, 2020, 27: 3025-3034.

[60] PradhanS, NautiyalV, DubeyRC. Antioxidant potential and molecular docking of bioactive compound of Camellia sinensis and Camellia assamica with cytochrome P450. Arch Microbiol, 2022, 204: 350.

[61] Erny SabrinaMN, RazaliM, MirfatAHS, Mohd ShukriMA. Antimicrobial activity and bioactive evaluation of Plectranthus amboinicus essential oil. Am J Res Commun, 2014, 2: 121-127

[62] Quintans-JúniorL, MoreiraJCF, PasqualiMAB, RabieSMS, PiresAS, SchröderR, et al. . Antinociceptive activity and redox profile of the monoterpenes (+)-camphene, p-cymene, and geranyl acetate in experimental models. ISRN Toxicol, 2013, 2013: 1-11.

[63] Dike-NdudimJN, AmadiCN, NdubuezeCW. Antimicrobial and phytochemical evaluation of Datura Stramonium (Jimsonweed) on selected microorganisms. Int J Sci Res Arch, 2021, 2: 245-256.

[64] HawarSN, TahaZK, HamiedAS, Al-ShmganiHS, SulaimanGM, ElsilkSE. Antifungal activity of bioactive compounds produced by the endophytic fungus Paecilomyces sp. (JN227071.1) against Rhizoctonia solani. Int J Biomater, 2023, 2023: 1-8.

[65] Brahma P, Baruah S. Investigation of phytochemical constituents, GC-MS, DPPH free radical scavenging assay, and mineral contents of Glochidion sphaerogynum (Mull. Arg.) Kurz bark extract. Plant Sci Today 2023. https://doi.org/10.14719/pst.2019.

[66] ImsongL, MuraliM. DNA barcoding, determination of bioactive compounds, antioxidant and anti-diabetic property in edible gastropod Brotia costula (Rafinesque, 1833) of Dimapur district, Nagaland. Int J Pharm Sci Res, 2022, 14: 1795-1804

[67] PhatangareND, DeshmukhKK, MuradeVD, NaikwadiPH, HaseDP, ChavhanMJ, et al. . Isolation and characterization of β-sitosterol from Justicia gendarussa burm. F.-an anti-inflammatory compound. Int J Pharmacogn Phytochem Res, 2017, 9: 1280-1287.

[68] JerezS, SierraL, de BrunoMP. 17-Octadecynoic acid improves contractile response to angiotensin II by releasing vasocontrictor prostaglandins. Prostaglandins Other Lipid Mediat, 2012, 97: 36-42.

Funding

Department of Biotechnology, Ministry of Science and Technology, India(DBT/2020/NEHU/1333)

RIGHTS & PERMISSIONS

Jiangnan University