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Plant Pathogenic Fungi and Oomycetes pp 39–51Cite as

Protocols for Investigating the Leaf Mycobiome Using High-Throughput DNA Sequencing

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

Abstract

High-throughput sequencing of taxon-specific loci, or DNA metabarcoding, has become an invaluable tool for investigating the composition of plant-associated fungal communities and for elucidating plant–fungal interactions. While sequencing fungal communities has become routine, there remain numerous potential sources of systematic error that can introduce biases and compromise metabarcoding data. This chapter presents a protocol for DNA metabarcoding of the leaf mycobiome based on current best practices to minimize errors through careful laboratory practices and validation.

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Acknowledgments

The authors are grateful to Maggie Wagner, Edward Barge, Kyle Gervers, Thomas Wolpert, and Wenbo Ma for helpful feedback in the development of this protocol. This research was supported by the Department of Energy Feedstock Genomics Award 219086 (PEB) and the National Science Foundation Science Engineering and Education for Sustainability Award 1314095 (PEB).

Author information

Authors and Affiliations

  1. Department of Biological Sciences, University of Memphis, Memphis, TN, USA

    Shawn P. Brown

  2. Department of Botany and Plant Pathology, Oregon State University, Corvallis, OR, USA

    Devin R. Leopold & Posy E. Busby

Authors
  1. Shawn P. Brown
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  2. Devin R. Leopold
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  3. Posy E. Busby
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Corresponding author

Correspondence to Posy E. Busby .

Editor information

Editors and Affiliations

  1. Department of Microbiology and Plant Pathology, University of California Riverside, Riverside, CA, USA

    Wenbo Ma

  2. Oregon State University, Corvallis, OR, USA

    Thomas Wolpert

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Brown, S.P., Leopold, D.R., Busby, P.E. (2018). Protocols for Investigating the Leaf Mycobiome Using High-Throughput DNA Sequencing. 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_4

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  • DOI: https://doi.org/10.1007/978-1-4939-8724-5_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8723-8

  • Online ISBN: 978-1-4939-8724-5

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