Abstract
Cephalotaxus mannii Hook f. is a rare medicinal plant used for leukemia treatment, however its fungal endophytes as a promising resource of pharmaceutical compounds remain poorly characterized. The present study is the first research on endophytic fungi associated with C. mannii collected in Vietnam and their potent biological activities. A total of 18 endophytic fungi were isolated; according to the results of morphological and internal transcribed spacer (ITS) sequence analyses, they belonged to six genera: Penicillium, Fusarium, Aspergillus, Diaporthe, Megasporoporia, and Trichoderma. Among these strains, the ethyl acetate extracts of Penicillium citrinum WDF8 and Fusarium perseae WDF12 showed remarkable antibacterial activity against at least 5 tested bacteria. Of note, only the WDF12 extract displayed potential cytotoxicity against A549 and MCF7 cancer cell lines with IC50 values at 13.4 ± 0.9 and 17.8 ± 2 µg/mL, respectively. Further cytotoxicity evaluation led to the identification of 10-deacetylbaccatin III-10-O-acetyl transferase (dbat) gene essential for the biosynthesis of the anti-cancer drug paclitaxel in the fungal strain WDF8. Both extracts also showed strong antioxidant activity against DPPH, hydroxyl, and superoxide anion radicals, attributed to a high level of polyphenols and flavonoids present. This study proved that endophytic fungi from C. mannii exhibit excellent antibacterial, anticancer, and antioxidant activities and may be promising candidates for the production of paclitaxel and new compounds.
Similar content being viewed by others
REFERENCES
Abdel-Fatah, S.S., El-Batal, A.I., El-Sherbiny, G.M., Khalaf, M.A., and El-Sayed, A.S., Production, bioprocess optimization and γ-irradiation of Penicillium polonicum, as a new Taxol producing endophyte from Ginko biloba, Biotechnol. Rep., 2021, vol. 30, p. e00623.
Carocho, M. and Ferreira, I.C., A review on antioxidants, prooxidants and related controversy: natural and synthetic compounds, screening and analysis methodologies and future perspectives, Food. Chem. Toxicol., 2013, vol. 51, pp. 15−25.
da Silva, M.H.R., Cueva-Yesquén, L.G., Júnior, S.B., Garcia, V.L., Sartoratto, A., de Angelis, D.F., and de Angelis, D.A., Endophytic fungi from Passiflora incarnata: an antioxidant compound source, Arch. Microbiol., 2020, vol. 202, pp. 2779−2789.
Das, A., Rahman, M.I., Ferdous, A.S., Amin, A., Rahman, M.M., Nahar, N., Uddin, M.A., Islam, M.R., and Khan, H., An endophytic basidiomycete, Grammothele lineata, isolated from Corchorus olitorius, produces paclitaxel that shows cytotoxicity, PLoS One, 2017, vol. 12, p. e0178612.
de Lima Souza, H.M., Sette, L.D., da Mota, A.J., do Nascimento Neto, J.F., Rodrigues, A., de Oliveira, T.B., de Oliveira, F.M., de Oliveira, L.A., dos Santos Barroso, H., and Zanotto, S.P., Filamentous fungi isolates of contaminated sediment in the Amazon region with the potential for benzo(a)pyrene degradation, Water Air Soil. Pollut., 2016, vol. 227, p. 431.
Dramae, A., Intaraudom, C., Bunbamrung, N., Boonyuen, N., Auncharoen, P., and Pittayakhajonwut, P., Antimicrobial tanzawaic acid derivatives from the endophytic Penicillium citrinum BCC71086, Tetrahedron, 2022, vols. 106−107, p. 132645.
Garyali, S., Kumar, A., and Reddy, M.S., Taxol production by an endophytic fungus, Fusarium redolens, isolated from Himalayan yew, J Microbiol Biotechnol., 2013, vol. 23, pp. 1372−1380.
Gautam, V.S., Singh, A., Kumari, P., Nishad, J.H., Kumar, J., Yadav, M., Bharti, R., Prajapati, P., and Kharwar, R.N., 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, vol. 204, p. 140.
Gonelimali, F.D., Lin, J., Miao, W., Xuan, J., Charles, F., Chen, M., and Hatab, S.R., Antimicrobial properties and mechanism of action of some plant extracts against food pathogens and spoilage microorganisms, Front Microbiol., 2018, vol. 9.
Kadaikunnan, S., Rejiniemon, T., Khaled, J.M., Alharbi, N.S., and Mothana, R., In-vitro antibacterial, antifungal, antioxidant and functional properties of Bacillus amyloliquefaciens, Ann. Clin. Microbiol. Antimicrob., 2015, vol. 14, p. 9.
Keller, N.P., Fungal secondary metabolism: regulation, function and drug discovery, Nat. Rev. Microbiol., 2019, vol. 17, pp. 167−180.
Kim, H., You, Y.H., Yoon, H., Seo, Y., Kim, Y.E., Choo, Y.S., Lee, I.J., Shin, J.H., and Kim, J.G., Culturable fungal endophytes isolated from the roots of coastal plants inhabiting korean East coast, Mycobiology, 2014, vol. 42, pp. 100−108.
Kumar, P., Singh, B., Thakur, V., Thakur, A., Thakur, N., Pandey, D., and Chand, D., Hyper-production of taxol from Aspergillus fumigatus, an endophytic fungus isolated from Taxus sp. of the Northern Himalayan region, Biotechnol. Rep., 2019, vol. 24, p. e00395.
Kumar, S., Stecher, G., and Tamura, K., MEGA7: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets, Mol. Biol. Evol., 2016, vol. 33, pp. 1870−1874.
Lee, D., Shim, S., and Kang, K., 4,6'-Anhydrooxysporidinone from Fusarium lateritium SSF2 induces autophagic and apoptosis cell death in MCF-7 breast cancer cells, Biomolecules, 2021, vol. 11.
Li, H. and Cui, B., Taxonomy and phylogeny of the genus Megasporoporia and its related genera, Mycologia, 2013, vol. 105, pp. 368−383.
Mohana, K.P., Zuehlke, S., Priti, V., Ramesha, B.T., Shweta, S., Ravikanth, G., Vasudeva, R., Santhoshkumar, T.R., Spiteller, M., and Uma Shaanker, R., Fusarium proliferatum, an endophytic fungus from Dysoxylum binectariferum Hook f., produces rohitukine, a chromane alkaloid possessing anti-cancer activity, A. van Leeuwenjoek, 2012, vol. 101, pp. 323−329.
Ngo, C.C., Nguyen, Q.H., Nguyen, T.H., Quach, N.T., Dudhagara, P., Vu, T.H.N., Le, T.T.X., Le, T.T.H., Do, T.T.H., Nguyen, V.D., Nguyen, N.T., and Phi, Q.-T., Identification of fungal community associated with deterioration of optical observation instruments of museums in Northern Vietnam, Appl. Sci., 2021, vol. 11, p. 5351.
Saithong, P., Panthavee, W., Stonsaovapak, S., and Congfa, L., Isolation and primary identification of endophytic fungi from Cephalotaxus mannii trees, Maejo Int. J. Sci. Technol., 2010, vol. 4, pp. 446−453.
Santos, C., Santos da Silva, B.N., Amorim Ferreira e Ferreira, A.F.T., Santos, C., Lima, N., and Silva Bentes, J.L.d., Fungal endophytic community associated with guarana (Paullinia cupana var. sorbilis): diversity driver by genotypes in the centre of origin, J. Fungi, 2020, vol. 6, p. 123.
Skehan, P., Storeng, R., Scudiero, D., Monks, A., McMahon, J., Vistica, D., Warren, J.T., Bokesch, H., Kenney, S., and Boyd, M.R., New colorimetric cytotoxicity assay for anticancer-drug screening, J. Natl. Cancer Inst., 1990, vol. 82, pp. 1107−1112.
Soliman, S.S. and Raizada, M.N.J.F.i.m., Darkness: a crucial factor in fungal taxol production, Front. Microbiol., 2018, vol. 9, p. 353.
Stierle, A., Strobel, G., and Stierle, D., Taxol and taxane production by Taxomyces andreanae, an endophytic fungus of Pacific yew, Science, 1993, vol. 260, pp. 214−216.
Stierle, A.A. and Stierle, D.B., Bioactive compounds from four endophytic Penicillium sp. of a northwest Pacific yew tree, Stu. Nat. Prod. Che., 2000, vol. 1, pp. 933−977.
Tang, Z., Wang, Y., Yang, J., Xiao, Y., Cai, Y., Wan, Y., Chen, H., Yao, H., Shan, Z., Li, C., and Wang, G., Isolation and identification of flavonoid-producing endophytic fungi from medicinal plant Conyza blinii H. Lév that exhibit higher antioxidant and antibacterial activities, PeerJ, 2020, vol. 8, p. e8978.
Toghueo, R.M.K., Bioprospecting endophytic fungi from Fusarium genus as sources of bioactive metabolites, Mycology, 2020, vol. 11, pp. 1−21.
Tsuda, M., Sasaki, M., Mugishima, T., Komatsu, K., Sone, T., Tanaka, M., Mikami, Y., Kobaya-shi, J.I., Scalusamides A−C, new pyrrolidine alkaloids from the marine-derived fungus Penicillium citrinum, J. Nat. Prod., 2005, vol. 68, pp. 273−276.
Vu, T.H.N., Pham, N.S., Le, P.C., Pham, Q.A., Quach, N.T., Do, T.T., Chu, H.H., and Phi, Q.T., Distribution, cytotoxicity, and antioxidant activity of fungal endophytes isolated from Tsuga chinensis (Franch.) Pritz. in Ha Giang province, Vietnam, Ann. Microbiol., 2022, vol. 72, pp. 1−12.
Yang, H.R., Hu, X.P., Jiang, C.J., Qi, J., Wu, Y.C., Li, W., Zeng, Y.J., Li, C.F., and Liu, S.X., Diversity and antimicrobial activity of endophytic fungi isolated from Cephalotaxus hainanensis Li, a well-known medicinal plant in China, Lett. Appl. Microbiol., 2015, vol. 61, pp. 484−490.
Zhang, P., Zhou, P.P., Jiang, C., Yu, H., and Yu, L.J., Screening of taxol-producing fungi based on PCR amplification from Taxus, Biotechnol., Lett., 2008, vol. 30, pp. 2119−2123.
ACKNOWLEDGMENTS
The authors would like to thank the support of VAST– Culture Collection of Microorganisms, Institute of Biotechnology, Vietnam Academy of Science and Technology (www.vccm.vast.vn).
Funding
This study was financially supported by Vietnam Academy of Science and Technology under Grant number TĐCNSH.05/20-22.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
The authors declare that they have no conflicts of interest.
This article does not contain any studies involving animals or human participants performed by any of the authors.
Supplementary Information
Rights and permissions
About this article
Cite this article
Vu, T.H., Quach, N.T., Le, P.C. et al. Bioprospecting Endophytic Fungi Isolated from Cephalotaxus mannii Hook f. as Prolific Sources of Antibacterial, Anticancer, and Antioxidant Agents. Microbiology 92, 284–292 (2023). https://doi.org/10.1134/S0026261722602834
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1134/S0026261722602834