Skip to main content
Log in

Molecular characterization and antimicrobial activity of endophytic fungi from coffee plants

  • Original Paper
  • Published:
World Journal of Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Endophytic filamentous fungi from coffee plants (Coffea arabica and C. robusta) deposited in the Brazilian Collection of Environmental and Industrial Microorganisms (CBMAI) were characterized taxonomically by using molecular tools and investigated concerning their antimicrobial activity against different human pathogenic bacteria. Thirty-seven out of 39 CBMAI strains investigated were identified to at least at genus level by ITS and rDNA D1/D2 sequencing and phylogenetic analyses. Bioactivity screening of fungal extracts against Salmonella choleraesuis (CBMAI 484), Staphylococcus aureus (CBMAI 485), Pseudomonas aeruginosa (CBMAI 489) and against four different Escherichia coli serotypes showed that 17 fungi inhibited at least one of the bacteria studied. The endophytic fungi Trichoderma harzianum (CBMAI 43), Guignardia sp. (CBMAI 69) and Phomopsis sp. (CBMAI 164) inhibited from four to five bacterial species, while five fungi were active against all pathogenic bacteria tested and were identified as Aspergillus versicolor (CBMAI 46), Fusarium oxysporum (CBMAI 53), Glomerella sp. (CBMAI 63) and Cladosporium spp. (CBMAI 64 and CBMAI 66). The Minimal Inhibitory Concentration (MIC) for the fungus extracts varied from 0.025 to 1.0 mg ml−1, demonstrating antimicrobial potential of some of these fungi.

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

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Aligiannis N, Kalpotzakis E, Mitaku S, Chinou IB (2001) Composition and antimicrobial activity of the essential oils of two Origanum species. J Agric Food Chem 40:4168–4170

    Article  CAS  Google Scholar 

  • Abd-Elsalam K, Aly IB, Abdel-Satar MA, Khalil MS, Verreet JA (2003) PCR identification of Fusarium genus on nuclear-ribosomal-DNA sequence data. Afr J Biotechnol 2:82–85

    Google Scholar 

  • Ablitz P, Fukushima K, Takizawa K, Nishimura K (2004) Identification of pathogenic dematiaceous fungi and related taxa based on large subunit ribosomal DNA D1/D2 domain sequence analysis. FEMS Immunol Med Microbiol 40:41–49

    Article  PubMed  CAS  Google Scholar 

  • Azevedo JL, Maccheroni WJ, Pereira JO, Araujo WL (2000) Endophytic microorganisms: a review on insect control and recent advances on tropical plants. Elect J Biotechnol 3 (www.ejbiotechnology.info/content/vol3/issue1/full/4/)

  • Bacon CW, White JF Jr (eds) (2000). Microbial endophytes. Marcel-Dekker, New York, 487 pp ISBN 0-8247-8831-1

    Google Scholar 

  • Brookman JL, Mennim G, Trinci APJ, Theodorou MK, Tuckwell DS (2000) Identification and characterization of anaerobic gut fungi using molecular methodologies based on ribosomal ITS1 and 18S rRNA. Microbiology 146:393–403

    PubMed  CAS  Google Scholar 

  • Ellof JN (1998) A sensitive and quick microplate method to determine the minimal inhibitory concentration of plant extracts for bacteria. Planta Medica 64:711–713

    PubMed  Google Scholar 

  • Ezra D, Hess WM, Strobel GA (2004) New endophytic isolates of Muscodor albus, a volatile-antibiotic-producing fungus. Microbiology 150:4023–4031

    Article  PubMed  CAS  Google Scholar 

  • Glienke-Blanco C (1999) Guignardia citricarpa Kiely: Análise genética cariotípica e interação com o hospedeiro. p 200. São Paulo: Tese de Doutorado ESALQ, Piracicaba

  • Hanlin RT, Menezes M (1996) Gêneros ilustrados de ascomicetos. p 274. UFRPe. Recife-PE

  • Kimura M (1980) A simple method for estimating evolutionary rate of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 16:111–120

    Article  PubMed  CAS  Google Scholar 

  • Kobayashi DY, Palumbo JD (2000) Bacterial endophytes and their effects on plants and uses in agriculture. In: Bacon CW, White JF (eds) Microbial endophytes. Marcel Deckker, New York ISBN 0-8247-8831-1

    Google Scholar 

  • Kumar S, Tamura K, Nei M (2004) MEGA3: Integrated software for Molecular Evolutionary Genetics Analysis and sequence alignment. Briefings Bioinform 5:150–153

    Article  CAS  Google Scholar 

  • LoBuglio KF, Taylor JW (1995) Phylogeny and PCR identification of the human pathogenic fungus Penicillium maeneffei. J Clin Microbiol 33:85–89

    PubMed  CAS  Google Scholar 

  • Nunes FV (2004) Isolamento e identificação de bactérias endofíticas do café (Coffea arabica e Coffea robusta) e seu potencial biotecnológico. Tese de Mestrado. Pós-graduação em Biotecnologia, Universidade de São Paulo, Brasil

  • Peintne U, Moser MM, Thomas KA, Manimohan (2003) First records of ectomycorrhizal Cortinarius species (Agaricales, Basidiomycetes) from tropical India and their phylogenetic position based on rDNA ITS sequences. Mycological Research 107:485–494

    Article  PubMed  CAS  Google Scholar 

  • Peláez F, Collado J, Arenal F, Basilio A, Cabello A, Matas MTD, Garcia JB, del Val AG, Gonzalez V, Gorrochategui J, Hernandez P, Martin I, Platas G, Vicente F (1998) Endophytic fungi from plants living on gypsum soils as a source of secondary metabolites with antimicrobial activity. Mycol Res 102:755–761

    Article  Google Scholar 

  • Peláez F, Cabello A, Platas G, Diez MT, del Val AG, Basilio A, Martan I, Vicente F, Bills GF, Giacobbe RA, Schwartz RE, Onishi JC, Meinz MS, Abruzzo GK, Flattery AM, Kong L, Kurtz MB (2000) The discovery of enfumafungin, a novel antifungal compound produced by an endophytic Hormonema species biological activity and taxonomy of the producing organisms. Systematic Appl Microbiol 23:333–343

    Google Scholar 

  • Radu S, Kqueen CY (2000) Preliminary screening of endophytic fungi from medicinal plants in malaysia for antimicrobial and antitumor activity. Malaysian J Med Sci 9:23–33

    Google Scholar 

  • Redecker D (2000) Specific PCR primers to identify arbuscular mycorrhizal fungi within colonized roots. Mycorrhiza 10:73–80

    Article  CAS  Google Scholar 

  • Rehder VLG, Machado ALM, Delarmelina C, Sartoratto A, Figueira GM, Duarte MCT (2004) Chemical composition and antimicrobial activity os essencial oil from Origanum applii e Origanum vulgare. Brazilian J Med Plants 6:67–71

    CAS  Google Scholar 

  • Sartoratto A, Machado ALM, Delarmelina C, Figueira GM, Duarte MCT, Rehder VLG (2004) Composition and antimicrobial activity of essential oils from aromatic plants used in Brazil. Brazilian J Microbiol 35:275–280

    CAS  Google Scholar 

  • Schabereiter-Gurtner C, Piñar G, Lubita W, Rölleke S (2001) Analysis of fungal communities on historical church window glass by denaturing gradient gel eletrophoresis and phylogenetic 18 rDNA sequence analysis. J Microbiol Methods 47:345–354

    Article  PubMed  CAS  Google Scholar 

  • Sette LD, Oliveira VM, Manfio GP (2005) Isolation and characterization of alachlor-degrading actinomycetes from soil. Antonie van Leeuwenhoek 87:81–89

    Article  PubMed  CAS  Google Scholar 

  • Singh BB, Zink D, Polishook J, Vaentino A, Shafiee D, Silverman K, Felock P, Teran A, Vilella D, Hazuda DJ, Lingham RB (1999) Structure and absolute stereochemistry of HIV−1 integraseinhinbitor integric acid, a novel eremophilanesequisterpenoid produced by a Xylaria sp. Tetrahedrone Lett 40:8775–8779

    Article  CAS  Google Scholar 

  • Smit E, Leeflang P, Glandorf B, va Elsas JD, Wernars K (1999) Analysis of fungal diversity in the eheat rhizosphere by sequencing of cloned PCR-amplified genes encoding 18S rRNA and temperature gradient gel electrophoresis. Appl Environ Microbiol 65:2614–2621

    PubMed  CAS  Google Scholar 

  • Souza AQL, Souza ADL, Astolfi Filho S, Belém Pinheiro ML, Sarquis MIM, Pereira JO (2004) Atividade antimicrobiana de fungos endofíticos isolados de plantas tóxicas da Amazônia: Palicourea longiflora (aubl.) rich e Strychnos cogens bentham. Acta Amazônica 34:185–195

    Google Scholar 

  • Sterflinger K, Prillinger H (2001) Molecular taxonomy and biodiversity of rock fungal communities in an urban environment (Vienna, Austria). Antonie van Leeuwenhoek 80:275–286

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Sutar R, David JK, Ghosh AK, Singhi S, Chakrabarti A, Bachhawat AK (2004) Comparison of ITS and IGS regions for strain typing of clinical and non-clinical isolates of Pichia anomala. J Med Microbiol 53:1–5

    Article  PubMed  CAS  Google Scholar 

  • Thompson JD, Higgins DG, Gibson TJ, Clustal W (1994) Improving the sensitiviy of rogressive multiple alignment through sequence weighting, positions-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680

    PubMed  CAS  Google Scholar 

  • van Elsas JD, Duarte GF, Keijzer-Wolters, Smit E (2000) Analysis of the dynamics of fungal communities in soil via fungal-specific PCR of soil DNA followed by denaturing gradient gel electrophoresis. J Microbiol Methods 43:133–151

    Article  PubMed  Google Scholar 

  • Wu Z, Tsumura T, Blonquist G, Wang X (2003) 18 rRNA gene variation among common airborne fungi and development of specific oligonucleotide probe for the detection of fungal isolates. Appl Environ Microbiol 69:5389–5397

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

This research was funded by Microbiology and Microbial Resource Divisions (CPQBA-UNICAMP). We thank Manuela da Silva (INCQS-FIOCRUZ) for the article criticisms and suggestions, Andressa Almeida Soares and Claudia Beatriz Afonso de Menezes for helping in preparation of extracts for antimicrobial assay, and Ana Paula Macedo Zibordi (CBMAI mycologist) for the supply of the morphologic characterization data.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to L. D. Sette.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sette, L.D., Passarini, M.R.Z., Delarmelina, C. et al. Molecular characterization and antimicrobial activity of endophytic fungi from coffee plants. World J Microbiol Biotechnol 22, 1185–1195 (2006). https://doi.org/10.1007/s11274-006-9160-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11274-006-9160-2

Keywords

Navigation