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Coniochaeta ligniaria: antifungal activity of the cryptic endophytic fungus associated with autotrophic tissue cultures of the medicinal plant Smallanthus sonchifolius (Asteraceae)

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Abstract

Few studies have addressed the presence and bioactivity of endophytic fungi living in plantlets growing under in vitro conditions. After unfruitful attempts to grow axenic shoot cultures of the medicinal plant Smallanthus sonchifolius (yacon) were made, healthy shoots grew on half strength Murashigue and Skoog media supplemented with 2.2 μM benzylaminopurine without sucrose. We isolated a fungus UM109 from these autotrophic tissue cultures and it was identified as Coniochaeta ligniaria using molecular, physiological and morphological methods. Dichloromethane extracts from C. ligniaria and its host S. sonchifolius exhibited antifungal activity against phytopathogenic fungi Colletotrichum acutatum, C. fragariae and C. gloeosporioides. Both extracts of C. ligniaria and S. sonchifolius were subjected to antifungal bioassay-directed fractionation using NMR spectroscopy and GC-FID analysis. Twelve antifungal fatty acids were identified and 8 out of the total were produced by the fungus and the plant including caproic, caprylic, myristic, palmitic, heptadecanoic, stearic, oleic and linoleic acids. Additionally, caproic, caprylic and palmitic acids were produced at high concentrations by the endophytic fungus and its host. The detection of these antifungal fatty acids produced by both C. ligniaria and S. sonchifolius suggests that these bioactive compounds may be partially responsible for the high resistance of S. sonchifolius to phytopathogenic fungal attacks. This finding also indicates the existence of an interesting chemical symbiosis between an endophytic fungus and its host. Furthermore, the isolation of C. ligniaria from tissue culture of S. sonchifolius demonstrates that plantlets growing in vitro as autotrophic cultures can shelter specific endophytic fungal communities. The use of autotrophic tissue cultures may become an important tool for studies on the taxonomy, ecology, evolution and biotechnological application of endophytes.

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Acknowledgements

This work received support of the Conselho Nacional of the Desenvolvimento Científico and Tecnológico (processo 200774/2011-5), Fundação of Amparo the Pesquisa of Minas Gerais (FAPEMIG), and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES/PGCI 036/13). The authors thank Ms. J.L. Robertson, Mrs. R. Pace, Mrs. Amber Reichley, Mr. Solomon Green III and Mrs. Marilyn Ruscoe for technical support. We also thank Dr. N. Tabanca and Dr. A. L. Cerdeira for valuable comments on the manuscript. The National Center for Natural Products Research, University of Mississippi is partially supported through a cooperative scientific agreement 58-6408-2-0009 from the USDA, Agricultural Research Service.

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Authors and Affiliations

  1. Laboratório de Sistemática e Biomoléculas de Fungos, Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, P. O. Box 486, 31270-901, Brazil

    Luiz H. Rosa

  2. EMBRAPA Meio Ambiente, P.O Box 69, Jaguariúna, 13820-000, SP, Brazil

    Sonia C. N. Queiroz

  3. National Center for Natural Products Research, Research Institute of Pharmaceutical Sciences, School of Pharmacy, University of Mississippi, University, MS, 38677, USA

    Rita M. Moraes & Natascha Techen

  4. Center for Water and Wetland Resources, Biological Field Station, University of Mississippi, 15 County Road 2078, Abbeville, MS, 38601, USA

    Rita M. Moraes

  5. Department of Medicinal Chemistry, University of Mississippi, MS, USA

    Xiaoning Wang

  6. USDA-ARS, National Products Utilization Research Unit (NPURU), University, MS, 38677, USA

    Zhiqiang Pan, Charles L. Cantrell & David E. Wedge

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  1. Luiz H. Rosa
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  2. Sonia C. N. Queiroz
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  3. Rita M. Moraes
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  7. Charles L. Cantrell
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  8. David E. Wedge
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Correspondence to Luiz H. Rosa or David E. Wedge.

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Rosa, L.H., Queiroz, S.C.N., Moraes, R.M. et al. Coniochaeta ligniaria: antifungal activity of the cryptic endophytic fungus associated with autotrophic tissue cultures of the medicinal plant Smallanthus sonchifolius (Asteraceae). Symbiosis 60, 133–142 (2013). https://doi.org/10.1007/s13199-013-0249-8

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