Skip to main content

Analysis of Secondary Metabolites from Plant Endophytic Fungi

  • Protocol
  • First Online:
Plant Pathogenic Fungi and Oomycetes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1848))

Abstract

Filamentous fungi produce a wide variety of secondary metabolites, which are recognized as a rich source of biomolecules with potential medicinal applications. According to chemical structures and biosynthetic pathways, secondary metabolites are mainly divided into four groups, including polyketide, nonribosomal peptides, alkaloids, and terpenes. Owing to their special living environment, plant endophytic fungi potentially produce many bioactive metabolites with diverse structural features. Some of these metabolites have been demonstrated to hold pharmaceutical and ecological significance. In order to identify bioactive compounds, different methods have been developed to isolate secondary metabolites from plant endophytic fungi. In this chapter, we describe the procedures for fungal fermentation, metabolite extraction, and analysis using high-performance liquid chromatography (HPLC). The endophytic fungus Pestalotiopsis fici, which was isolated from the healthy branches of Camellia sinensis (Theaceae), produces 88 secondary metabolites including 70 new natural products. Combining 74 secondary metabolite clusters revealed by genome sequencing analysis, P. fici is used as an example for analysis of secondary metabolites. The procedures could also be adopted for other fungi after small modifications.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Hoffmeister D, Keller NP (2007) Natural products of filamentous fungi: enzymes, genes, and their regulation. Nat Prod Rep 24:393–416

    Article  CAS  PubMed  Google Scholar 

  2. Beck J, Ripka S, Siegner A, Schiltz E, Schweizer E (1990) The multifunctional 6-methylsalicylic acid synthase gene of Penicillium patulum. Its gene structure relative to that of other polyketide synthases. Eur J Biochem 192:487–498

    Article  CAS  PubMed  Google Scholar 

  3. Liu J, Farmer JD Jr, Lane WS, Friedman J, Weissman I, Schreiber SL (1991) Calcineurin is a common target of cyclophilin-cyclosporin a and FKBP-FK506 complexes. Cell 66:807–815

    Article  CAS  PubMed  Google Scholar 

  4. Martín JF, Ullán RV, García-Estrada C (2010) Regulation and compartmentalization of β-lactam biosynthesis. Microb Biotechnol 3:285–299

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Shweta S, Zuehlke S, Ramesha BT, Priti V, Mohana KP, Ravikanth G et al (2010) Endophytic fungal strains of Fusarium solani, from Apodytes dimidiata E. Mey. ex Arn (Icacinaceae) produce camptothecin, 10-hydroxycamptothecin and 9-methoxycamptothecin. Phytochemistry 71:117–122

    Article  CAS  PubMed  Google Scholar 

  6. Panaccione DG, Coyle CM (2005) Abundant respirable ergot alkaloids from the common airborne fungus Aspergillus fumigatus. Appl Environ Microbiol 71:3106–3111

    Google Scholar 

  7. Daum M, Herrmann S, Wilkinson B, Bechthold A (2009) Genes and enzymes involved in bacterial isoprenoid biosynthesis. Curr Opin Chem Biol 13:180–188

    Article  CAS  PubMed  Google Scholar 

  8. Strobel G, Daisy B, Castillo U, Harper J (2004) Natural products from endophytic microorganisms. J Nat Prod 67:257–268

    Article  CAS  PubMed  Google Scholar 

  9. Strobel G, Daisy B (2003) Bioprospecting for microbial endophytes and their natural products. Microbiol Mol Biol Rev 67:491–502

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Stierle A, Strobel G, Stierle D (1993) Taxol and taxane production by Taxomyces andreanae, an endophytic fungus of Pacific yew. Science 260:214–216

    Article  CAS  PubMed  Google Scholar 

  11. Metz AM, Haddad A, Worapong J, Long DM, Ford EJ, Hess WM, Strobel GA (2000) Induction of the sexual stage of Pestalotiopsis microspora, a taxol-producing fungus. Microbiology 146:2079–2089

    Article  CAS  PubMed  Google Scholar 

  12. Wilkinson HH, Siegel MR, Blankenship JD, Mallory AC, Bush LP, Schardl CL (2000) Contribution of fungal loline alkaloids to protection from aphids in a grass-endophyte mutualism. Mol Plant-Microbe Interact 13:1027–1033

    Article  CAS  PubMed  Google Scholar 

  13. Keller NP, Turner G, Bennett JW (2005) Fungal secondary metabolism—from biochemistry to genomics. Nat Rev Microbiol 3:937–947

    Article  CAS  PubMed  Google Scholar 

  14. Galagan JE, Calvo SE, Cuomo C, Ma LJ, Wortman JR, Batzoglou S et al (2005) Sequencing of Aspergillus nidulans and comparative analysis with A. fumigatus and A. oryzae. Nature 438:1105–1115

    Google Scholar 

  15. Dean RA, Talbot NJ, Ebbole DJ, Farman ML, Mitchell TK, Orbach MJ et al (2005) The genome sequence of the rice blast fungus Magnaporthe grisea. Nature 434:980–986

    Article  CAS  PubMed  Google Scholar 

  16. Liu L (2011) Bioactive metabolites from the plant endophyte Pestalotiopsis fici. Mycology 2:37–45

    Article  CAS  Google Scholar 

  17. Wang X, Zhang X, Liu L, Xiang M, Wang W, Sun X et al (2015) Genomic and transcriptomic analysis of the endophytic fungus Pestalotiopsis fici reveals its lifestyle and high potential for synthesis of natural products. BMC Genomics 16:28

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Liu L, Li Y, Li L, Cao Y, Guo L, Liu G, Che Y (2013) Spiroketals of Pestalotiopsis fici provide evidence for a biosynthetic hypothesis involving diversified Diels-Alder reaction cascades. J Org Chem 78:2992–3000

    Article  CAS  PubMed  Google Scholar 

  19. Liu L, Liu S, Jiang L, Chen X, Guo L, Che Y (2008) Chloropupukeananin, the first chlorinated pupukeanane derivative, and its precursors from Pestalotiopsis fici. Org Lett 10:1397–1400

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Gang Liu .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Liu, J., Liu, G. (2018). Analysis of Secondary Metabolites from Plant Endophytic Fungi. In: Ma, W., Wolpert, T. (eds) Plant Pathogenic Fungi and Oomycetes. Methods in Molecular Biology, vol 1848. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8724-5_3

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-8724-5_3

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8723-8

  • Online ISBN: 978-1-4939-8724-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics