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Plant Pathogenic Fungi and Oomycetes pp 139–150Cite as

Host-Induced Gene Silencing (HIGS) for Elucidating Puccinia Gene Function in Wheat

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

Abstract

Biotrophic fungi (Puccinia spp.) cause devastating diseases of wheat and other cereal species globally. The function of large repertories of genes from Puccinia spp. still needs to be discovered to understand the infection process of these obligate parasites, eventually to protect plants from rust diseases. Functional analysis of targeted genes is challenging due to the inherent difficulties with culturing the fungus and transforming the host. RNA interference (RNAi) is a conserved gene regulation process in eukaryotes and known to be a powerful genetic tool in plant biotechnology. More recently, host-induced gene silencing (HIGS) has been developed to assess pathogen gene function in plants. HIGS is an RNAi-based process where double stranded RNA (dsRNA) homologous to a pathogen gene can be expressed in a plant to induce targeted silencing of the pathogen gene. Here we described a detailed HIGS protocol for functional analysis of rust genes from Puccinia species in cereals. As an example we describe an experiment silencing the tryptophan 2-monooxygenase gene (Pgt-IaaM) from Puccinia graminis f. sp. tritici (Pgt) that is involved in virulence to wheat.

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Acknowledgments

This work was funded by National Institute of Food and Agriculture to S.H. (Grant No. 2010-65108-20568).

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Authors and Affiliations

  1. Department of Plant Pathology, Washington State University, Pullman, WA, USA

    Chuntao Yin & Scot H. Hulbert

Authors
  1. Chuntao Yin
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  2. Scot H. Hulbert
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Corresponding author

Correspondence to Scot H. Hulbert .

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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Yin, C., Hulbert, S.H. (2018). Host-Induced Gene Silencing (HIGS) for Elucidating Puccinia Gene Function in Wheat. 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_12

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  • DOI: https://doi.org/10.1007/978-1-4939-8724-5_12

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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

  • eBook Packages: Springer Protocols

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