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Plant Fungal Pathogens pp 219–236Cite as

The Application of Laser Microdissection to Profiling Fungal Pathogen Gene Expression in planta

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 835))

Abstract

Laser microdissection (LM) of plant tissues infected with a fluorescent protein-tagged fungus is a useful method for obtaining samples highly enriched in fungal RNA for downstream analysis such as hybridization to a microarray. This paper outlines the requirements for successful LM of infected tissues and details a set of protocols for (1) preparing and sectioning infected tissue samples under conditions that preserve both RNA integrity and cytological features; (2) capturing fungal structures via LM; and (3) extraction and amplification of transcripts for further analysis.

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Acknowledgments

We thank K. Ohtsu and P. Schnabel for discussions of LM technique, and Barbara Valent for stimulating discussions. We also thank Xiang Tang, Xiao-Wei Zhang, Zhi-Yong Zhang, Dong Zhang, and Ting-Lu Yuan for help in organizing the lab protocols. Wei-Hua Tang’s research is supported by the Ministry of Science and Technology of China (grant nos. 2007CB108700 and 2006CB101901).

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Authors and Affiliations

  1. National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China

    Wei-Hua Tang & Yan Zhang

  2. Department of Genetics, Development and Cell Biology, Iowa State University, Ames, IA, USA

    Jon Duvick

Authors
  1. Wei-Hua Tang
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  2. Yan Zhang
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  3. Jon Duvick
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Corresponding author

Correspondence to Jon Duvick .

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Editors and Affiliations

  1. Northern Crops Science Laboratory, USDA-ARS, North 18th Street 1307, Fargo, 58105-5677, North Dakota, USA

    Melvin D. Bolton

  2. Lab. Phytopathology, Wageningen University, Droevendaalsesteeg 1, Wageningen, 6708 PB, Netherlands

    Bart P.H.J. Thomma

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Tang, WH., Zhang, Y., Duvick, J. (2012). The Application of Laser Microdissection to Profiling Fungal Pathogen Gene Expression in planta . In: Bolton, M., Thomma, B. (eds) Plant Fungal Pathogens. Methods in Molecular Biology, vol 835. Humana Press. https://doi.org/10.1007/978-1-61779-501-5_14

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  • DOI: https://doi.org/10.1007/978-1-61779-501-5_14

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-500-8

  • Online ISBN: 978-1-61779-501-5

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