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Morphological and molecular characterization of Tulasnella spp. fungi isolated from the roots of Epidendrum secundum, a widespread Brazilian orchid

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Abstract

Tulasnella spp. are the main fungal symbionts of Brazilian Epidendrum orchids. The taxonomy of these fungi is largely based on ITS rDNA similarity, but culture dependent techniques are still essential to establish the true biological entity of the mycobiont. The aim of this study was to characterize morphologically and molecularly 16 Tulasnella spp. fungi isolated from three different populations of E. secundum and to test the coincidences between morphological and molecular characterization. Two uninucleate rhizoctonia fungi, obtained from Oncidium barbaceniae, and two phytopathogenic isolates were included as outgroups. Qualitative and quantitative morphological characteristics were analyzed using multivariate statistics and were able to distinguish Ceratobasidium, Tulasnella and Thanatephorus genera and separate the isolates of Tulasnella spp. into two groups. Analysis of RAPD (Random Amplified Polymorphic DNA) and ITS rDNA sequences validated the morphological data. Symbionts of O. barbaceniae presented identity to ITS sequences of Ceratobasidium genus, while E. secundum isolates presented identity to two species of Tulasnella. We observed homogeneity among Tulasnella spp. obtained from a single population and from neighboring populations, but there was higher variability among isolates obtained from populations of regions that were farther apart. Morphological data associated with multivariate statistics proved to be a useful tool in the multi-level taxonomy of these orchid-associated fungi and in estimating the diversity of orchid mycorrhizal fungi.

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Acknowledgments

The authors thank the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and the Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) for financial support of project and for the PhD scholarship of the first author; the Instituto Estadual de Florestas (IEF) for granting us permission to collect material in the conservation area Serra do Brigadeiro, Minas Gerais; Gilmar Valente for his help during collections; two anonymous referees and Manuscript Editor of Symbiosis for improvements and advice.

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Authors and Affiliations

  1. Instituto de Ciências Biológicas e da Saúde, Universidade Federal de Viçosa, Campus de Rio Paranaíba, MG230, Km 7, Caixa Postal 22, CEP 38810-000, Rio Paranaíba, MG, Brazil

    Marlon Corrêa Pereira

  2. Departamento de Imunologia e Microbiologia Veterinária, Instituto de Veterinária, Universidade Federal do Rio de Janeiro, BR 465, Km 7, CEP 23890-000, Seropedica, RJ, Brazil

    Irene da Silva Coelho

  3. Departamento de Ciência do Solo, Escola Superior de Agricultura “Luiz de Queiroz”, Universidade de São Paulo, Avenida Pádua Dias, 11, CEP 13418-900, Piracicaba, SP, Brazil

    Rafael Borges da Silva Valadares

  4. Departamento de Microbiologia, Universidade Federal de Viçosa, Viçosa, Avenida Peter Henry Rolfs, s/n, CEP 36570-000, Viçosa, MG, Brazil

    Sabrina Feliciano Oliveira, Melissa Bocayuva, Elza Fernandes Araújo & Maria Catarina Megumi Kasuya

  5. Departamento de Fitopatologia, Universidade Federal de Viçosa, Avenida Peter Henry Rolfs, s/n, CEP 36570-000, Viçosa, MG, Brazil

    Olinto Liparini Pereira

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  1. Marlon Corrêa Pereira
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  2. Irene da Silva Coelho
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  3. Rafael Borges da Silva Valadares
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  4. Sabrina Feliciano Oliveira
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  5. Melissa Bocayuva
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  6. Olinto Liparini Pereira
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  7. Elza Fernandes Araújo
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  8. Maria Catarina Megumi Kasuya
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Correspondence to Marlon Corrêa Pereira.

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Pereira, M.C., da Silva Coelho, I., da Silva Valadares, R.B. et al. Morphological and molecular characterization of Tulasnella spp. fungi isolated from the roots of Epidendrum secundum, a widespread Brazilian orchid. Symbiosis 62, 111–121 (2014). https://doi.org/10.1007/s13199-014-0276-0

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  • DOI: https://doi.org/10.1007/s13199-014-0276-0

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