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Virus-Induced Gene Silencing (VIGS) for Functional Characterization of Disease Resistance Genes in Barley Seedlings

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Barley

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1900))

Abstract

With the recent advances in sequencing technologies, many studies are generating lists of candidate genes associated with specific traits. The major bottleneck in functional genomics is the validation of gene function. This is achieved by analyzing the effect of either gene silencing or overexpression on a specific phenotypic or biochemical trait. This usually requires the generation of stable transgenic plants and this can take considerable time. Therefore any technique that expedites the validation of gene function is of particular benefit in cereals, including barley. One such technique is Virus-Induced Gene Silencing (VIGS), which evokes a natural antiviral defense mechanism in plants. VIGS can be used to downregulate gene expression in a transient manner, but long enough to determine its effects on a specific phenotype. It is particularly useful for screening candidate genes and selecting those with potential for disease control. VIGS based on Barley Stripe Mosaic Virus (BSMV) is a powerful and efficient tool for the analysis of gene function in cereals. Here we present a BSMV VIGS protocol for simple and robust gene silencing in barley and describe it to evaluate the role of the hormone receptor BRI1 (Brassinosteroid Insensitive 1) in barley leaf resistance to Fusarium infection.

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Acknowledgments

This work was supported by the Science Foundation Ireland research fund (14/IA/2508) and Department of Agriculture Research Stimulus Grant RSF 11/S/103.

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Authors and Affiliations

  1. School of Biology and Environmental Science and Earth Institute, College of Science, University College Dublin, Dublin, Ireland

    Alexandre Perochon & Fiona M. Doohan

  2. SPCL, USDA/ARS Beltsville Agricultural Research Center, Beltsville, MD, USA

    Shahin S. Ali

  3. Crop Production and Pest Control Research Unit, USDA-ARS, West Lafayette, IN, USA

    Steven R. Scofield

  4. Department of Agronomy, Purdue University, West Lafayette, IN, USA

    Steven R. Scofield

  5. Department of Plant, Food, and Environmental Sciences, Dalhousie University, Truro, NS, Canada

    Lokanadha R. Gunupuru

Authors
  1. Lokanadha R. Gunupuru
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  2. Alexandre Perochon
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  3. Shahin S. Ali
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  4. Steven R. Scofield
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  5. Fiona M. Doohan
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Corresponding author

Correspondence to Fiona M. Doohan .

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Editors and Affiliations

  1. Crop Transformation Group, Department of Crop Genetics, John Innes Centre, Norwich, UK

    Wendy A. Harwood

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Gunupuru, L.R., Perochon, A., Ali, S.S., Scofield, S.R., Doohan, F.M. (2019). Virus-Induced Gene Silencing (VIGS) for Functional Characterization of Disease Resistance Genes in Barley Seedlings. In: Harwood, W. (eds) Barley. Methods in Molecular Biology, vol 1900. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8944-7_7

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  • DOI: https://doi.org/10.1007/978-1-4939-8944-7_7

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8942-3

  • Online ISBN: 978-1-4939-8944-7

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