Journal of Plant Molecular Breeding

Molecular and morphological identification of endophytic fungi isolated from aerial parts of yew tree

Document Type : Original research paper

Authors

1 Department of Horticultural Sciences, Gorgan University of Agricultural Sciences and Natural Resources ,

2 Gorgan University of Agricultural Sciences and Natural resources

3 Department of Horticulture Sciences, Faculty of Plant Production, Gorgan University of Agricultural Sciences and Natural Resources, Iran

4 Department of Plant Protection, Faculty of Plant Production, Gorgan University of Agricultural Sciences and Natural Resources, Iran.

Abstract

Plants are important parts of our diet; which, in addition to the primary metabolites, also produce a wide range of secondary metabolites. Endophytic fungi are among these microorganisms that not only do not cause infection and disease in the host plant but also play a role in the production or increase of some secondary metabolites and defense responses to biotic and abiotic stresses in the plant. Yew is one of the native trees of Iran and has medicinal value and has important fungal endophytes. Isolation and characterization of endophytic fungi in this plant can be important in order to discover new species with the potential to produce taxol. This study was carried out to isolate and identify molecularly and morphologically some fungal endophytes in the laboratory. In order to investigate the presence of ITS gene in fungi, ITS5 and ITS4 specific primers were used. 81 endophytic fungi were extracted from yew seedlings and eight samples were selected for molecular tests. The results of examining the presence of the ITS showed that all the extracted fungi gave a band in the desired region. Based on molecular and morphological studies, eight strains of identified endophytic fungi belonged to the genera Purpureocillium, Akanthomyces, Fusarium, Phomopsis and Colletotrichum. Therefore, the identification of more endophytic species can help to discover useful microorganisms in the production of secondary metabolites such as taxol.

Keywords

[1] Aharwal RP., Kumar S, Sandhu SS. 2014. Isolation and antibacterial property of endophytic fungi isolated from Indian medicinal plant Calotropis procera (Linn.) R.Br. World J Pharm Pharm Sci, 3: 678-691.
[2] Alam B, Li J, Ge Q, Alam Khan M, Gong J, Mehmood Sh, Yuan Y, Gong W. 2021. Endophytic Fungi: From Symbiosis to Secondary Metabolite Communications or Vice Versa? Front Plant Sci, 12: 791033.
[3] Arnold AE, Maynard A, Gilbert GS, Coley PD, Kursar TA. 2000. Are tropical fungal endophytes hyperdiverse? Ecol Lett, 3: 267-274.
[4] Arora P, Wani ZA, Ahmad T, Sultan P, Gupta S, Riyaz-Ul-Hassan S. 2019. Community structure, spatial distribution, diversity and functional characterization of culturable endophytic fungi associated with Glycyrrhiza glabra L. Fungal Biol, 123(5): 373-383.
[5] Begum N, Qin C, Ahanger MA, Raza S, Khan MI, Ahmed N, Ashraf M, Zhang L. 2019. Role of arbuscular mycorrhizal fungi in plant growth regulation: implications in abiotic stress tolerance. Front Plant Sci, 10:1068.
[6] Caruso M, Colombo AL, Fedeli L, Pavesi A, Quaroni S, Saracchi M. 2000. Isolation of endophytic fungi and actinomycetes taxane producers. Ann Microbiol, 50(1): 3-13.
[7] Cui R, Lu X, Chen X, Malik WA, Wang D, Wang J, Wang Sh, Guo L, Chen C, Wang X, Wang X, Dai M, Ye W. 2021. A novel raffinose biological pathway is observed by symbionts of cotton=Verticillium dahliae to improve salt tolerance genetically on cotton. J Agron Crop Sci, 207: 956-969.
[8] Ding B, Yuan J, Huang X, Wen W, Zhu X, Liu Y, Li H, Lu Y, He L, Tan H, She Z. 2013. New dimeric members of the phomoxanthone family: phomolactonexanthones A, B and deacetylphomoxanthone C isolated from the fungus Phomopsis sp. Mar Drugs, 11(12): 4961-4972.
[9] El Enshasy HA, Hanapi SZ, Malek RA, Abdelgalil SA, Leng OM. 2019. Endophytic fungi: the desired biostimulants for essential oil production. In: Fungal Biology. Advances in Endophytic Fungal Research. Pp: 211-232.
[10] El-Bialy HA, El-Bastawisy HS. 2020. Elicitors stimulate paclitaxel production by endophytic fungi isolated from ecologically altered Taxus baccata. J Radiat Res Appl Sci J, 13: 79-87.
[11] Gamboa MA, Bayman P. 2001. Communities of endophytic fungi in leaves of a tropical timber tree (Guarea guidonia: Meliaceae). Biotropical, 33(2): 352-360.
[12] Hastuti US, Al Asna PM, Rahmawati D. 2018. Histologic Observation, Identification, and Secondary Metabolites Analysis of Endophytic Fungi Isolated from a Medicinal Plant, Hedychium acuminatum Roscoe. AIP Conf Proc, 2002: 020070.
[13] Heinz KM, Harding PA, Ek-Ramos MJ, Hernandez H, Krauter PC, Sword GA. 2018. Fungal endophytes in Knock out Rose and performance effects of entomopathogens on marigold and zinnia. Hortic Sci, 53 (12): 1791-1798.
[14] Köhl J, Kolnaar R, Ravensberg WJ. 2019. Mode of action of microbial biological control agents against plant diseases: Relevance beyond efficacy. Front Plant Sci, 10: 845.
[15] Kumara PM, Zuehlke S, Priti V, Ramesha B, Shweta S, Ravikanth G, Vasudeva R, Santhoshkumar TR, Spiteller M, Shaanker R. 2012. Fusarium proliferatum, an endophytic fungus from Dysoxylum binectariferum Hook.f, produces rohitukine, a chromane alkaloid possessing anti-cancer activity. Antonie van Leeuwenhoek, 101: 323-329.
[16] Kuriakose GC, Arathi BP, Lakshmanan MD, Jiby MV, Gudde RS, Jayabhaskaran C. 2020. Sub-acute Toxicity Assessment of Taxol Isolated from Fusarium Solani, an Endophytic Fungus of Taxus Brevifolia, in Wistar Rats and Analyzing Its Cytotoxicity and Apoptotic Potential in Lung Cancer Cells. Origin Res, 10: 538865.
[17] Labossiere AW, Thompson DF. 2018. Clinical Toxicology of Yew Poisoning. Ann Pharmacother, 52(6): 591-599.
[18] Lenta BN, Ngatchou J, Frese M, Ladoh-Yemeda F, Voundi S, Nardella F, Michalek C, Wibberg D, Ngouela S, Tsamo E, Kaiser M, Kalinowski J, Sewald N. 2016. Purpureone, an antileishmanial ergochrome from the endophytic fungus Purpureocillium lilacinum. Z Naturforsch, 71(11) b: 1159-1167.
[19] Luangsa-Ard J, Houbraken J, van Doorn T, Hong SB, Borman AM, Hywel-Jones NL, Samson RA. 2011. Purpureocillium, a new genus for the medically important Paecilomyces lilacinus. FEMS Microbiol Lett, 321(2): 141-149.
[20] Mehta P, Sharma R, Putatunda C, Walia A. 2019. Endophytic fungi: role in phosphate solubilization. In Advances in Endophytic Fungal Research: Present Status and Future Challenges, ed. Singh, B.P. pp: 183-209.
[21] Morales-Sánchez V, Fe Andrés M, Díaz CE, González-Coloma A. 2020. Factors affecting the metabolite productions in endophytes: biotechnological approaches for production of metabolites. Curr Med Chem, 27(11): 1855-1873.
[22] Mosaddegh A. 2001. Yew tree. Ecol, 27(28): 73-74. (In Persian)
[23] Murray M, Thompson W. 1980. Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res, 8(19): 4321-4325.
[24] Naik BS. 2019. Developments in taxol production through endophytic fungal biotechnology: a review. Orient Pharm Exp Med, 19(1): 1-13.
[25] Oberhofer M, Wackerlig J, Zehl M, Buyuk H, Jian Cao J, Prado-Roller A, Urban E, Zotchev SB. 2021. Endophytic Akanthomyces sp. LN303 from Edelweiss Produces Emestrin and Two New 2- Hydroxy-4-Pyridone Alkaloids. ACS Omega, 6: 2184-2191.
[26] Omidbaigi R. 2010. Production and Processing of Medicinal Plants. Astan Qods Razavi Publications. 423 pp. (In Persian)
[27] Pandey KB, Rizvi SI. 2009. Plant polyphenols as dietary antioxidants in human health and disease. Oxid Med Cell Longev, 2(5): 270-278.
[28] Patil RH, Patil MP, Maheshwar V. 2016. Bioactive secondary metabolites from endophytic fungi: A review of biotechnological production and their potential applications. Stud Nat Prod Chem, 49: 189-205.
[29] Poveda J, Eugui D, Abril-Urías P, Velasco P. 2021. Endophytic fungi as direct plant growth promoters for sustainable agricultural production. Symbiosis, 85: 1-19.
[30] Raja A, Miller N, Pearce J, Oberlies H. 2017. Fungal identification using molecular tools: A primer for the natural products research community. Nat Prod J, 80: 756-770.
[31] Rivera-Orduna FN, Suarez-Sanchez RA, Flores-Bustamante ZR, Gracida-Rodriquez JN, Flores-Cotera LB. 2011. Diversity of endophytic fungi of Taxus globosa (Mexican yew). Fungal Divers, 47: 65-74.
[32] Salehi M, Moieni A, Safaie N, Farhadi S. 2020. Whole fungal elicitors boost paclitaxel biosynthesis induction in Corylus avellana cell culture. PloS One, 15(7): e0236191.
[33] Schulz B, Wanke U, Draeger S. 1993. Endophytes from herbaceous and shrubs: effectiveness of surface sterilization method. Mycol Res, 97: 1447-1450.
[34] Tazik Z, Rahnama K, Iranshahi M, White JF, Soltanloo H. 2020. A new species of Pithoascus and first report of this genus as endophyte associated with Ferula ovina. Mycoscience, 61: 145-150.
[35] Valko M, Leibfritz D, Moncol J, Cronin MTD, Mazur M, Telser J. 2007. Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol, 39: 44-84.
[36] Wang CJ, Wang YZ, Chu ZH, Wang PS, Liu BY, Li BY, Yu XL, Luan BH. 2020. Endophytic Bacillus amyloliquefaciens YTB1407 elicits resistance against two fungal pathogens in sweet potato (Ipomoea batatas (L.) Lam.). J Plant Physiol, 253: 153260.
[37] Wani MC, Taylor HL, Wall ME, Coggon P, McPhail AT. 1971. Plant antitumor agents. VI. Isolation and structure of taxol, a novel antileukemic and antitumor agent from Taxus brevifolia. Journal of the J Am Chem Soc, 93(9): 2325-2327.
[38] White TJ, Bruns TD, Lee SB, Taylor JW. 1990. Amplification and direct sequencing of fungal ribosomal RNA Genes for phylogenetics. In book: PCR - Protocols and Applications - A Laboratory Manual. Pp: 315-322.
[39] Xiong ZQ, Yang YY, Zhao N, Wang Y. 2013. Diversity of endophytic fungi and screening of fungal paclitaxel producer from Anglojap yew, Taxus × media. BMC Microbiol, 13: 71.
[40] Zhang P, Zhou PP, Yu LJ. 2009. An Endophytic taxol-producing fungus from Taxus media, Cladosporium cladosporioides MD2. Curr Microbiol, 59(3): 227-232.
[41] Zhao J, Fu Y, Luo M, Zu Y, Wang W, Zhao C, Gu C. 2012. Endophytic fungi from pigeon pea (Cajanus cajan (L.) Mill sp.) produce antioxidant cajaninstilbene acid. J Agric Food Chem, 60: 4314-4319.
Journal of Plant Molecular Breeding
Volume 9, Issue 2
December 2021
Pages 35-42
  • Receive Date: 19 September 2022
  • Revise Date: 17 October 2022
  • Accept Date: 25 October 2022
  • First Publish Date: 25 October 2022