Orchid Mycorrhizal Fungi: Isolation and Identification Techniques

  • Lawrence W. ZettlerEmail author
  • Laura L. Corey
Part of the Springer Protocols Handbooks book series (SPH)


In light of the importance of mycorrhizal fungi to the orchid life cycle from an evolutionary and ecological context, it is essential that conservationists adopt protocols to isolate, identify, and safeguard important fungal strains in this age of extinction. The purpose of this chapter is to summarize some of the methods used by researchers worldwide to isolate and identify peloton-forming fungi in the Rhizoctonia complex from temperate terrestrials and tropical epiphytes alike. The chapter begins with a summary of preparatory background information leading up to fungal isolation, including terminology, how to spot and identifying small seedlings in the field, how to avoid “senile populations,” collection permits, and viable transport of material. The second part of the chapter focuses on fungal isolation techniques, namely field collecting, peloton assessment, surface sterilization, and isolation from roots and protocorms. Maintenance of fungi through subculturing and storage is also discussed. The chapter concludes with methods aimed at provisional and precise fungal identification using cultural characteristics and modern molecular techniques, respectively, with emphasis on the Tulasnellaceae, Ceratobasidiaceae, and Sebacinaceae.

Key words

Seedlings Field collection Tulasnellaceae Ceratobasidiaceae Sebacinaceae Epiphytic and terrestrial orchids 



We kindly thank Michael E. Kane (University of Florida) for providing images of Tulasnella and Ceratobasidium in culture. The long-standing technical support of Andrew L. Stice (Illinois College) is much appreciated. We also thank our collaborators at the Royal Botanic Gardens, Kew (Viswambharan Sarasan, Kazutomo Yokoya, Jonathan Kendon), for the opportunity to collect samples in Madagascar that significantly enhanced this chapter. This chapter would not have been written without the expertise and guidance provided by Hanne N. Rasmussen (University of Copenhagen) for which we are grateful.


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Authors and Affiliations

  1. 1.Department of BiologyIllinois CollegeJacksonvilleUSA

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