Specific Isolation of RNA from the Grape Powdery Mildew Pathogen Erysiphe necator, an Epiphytic, Obligate Parasite

Authors

  • Lance Cadle-Davidson,

    1. Authors’ addresses: USDA-ARS, Grape Genetics Research Unit, 630 W. North St., Geneva
    2. Department of Plant Pathology and Plant Microbe Biology, Cornell University, Geneva, NY 14456, USA (correspondence to Lance Cadle-Davidson. E-mail: Lance.CadleDavidson@ars.usda.gov)
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      Equal first authors. Mention of trade names or commercial products is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the US Department of Agriculture.

  • Laura Wakefield,

    1. Department of Plant Pathology and Plant Microbe Biology, Cornell University, Geneva, NY 14456, USA (correspondence to Lance Cadle-Davidson. E-mail: Lance.CadleDavidson@ars.usda.gov)
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    • *

      Equal first authors. Mention of trade names or commercial products is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the US Department of Agriculture.

  • Robert C. Seem,

    1. Department of Plant Pathology and Plant Microbe Biology, Cornell University, Geneva, NY 14456, USA (correspondence to Lance Cadle-Davidson. E-mail: Lance.CadleDavidson@ars.usda.gov)
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  • David M. Gadoury

    1. Department of Plant Pathology and Plant Microbe Biology, Cornell University, Geneva, NY 14456, USA (correspondence to Lance Cadle-Davidson. E-mail: Lance.CadleDavidson@ars.usda.gov)
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USDA-ARS, Grape Genetics Research Unit, Geneva, USA

Abstract

RNA expression profiling of obligately parasitic plant microbes is hampered by the requisite interaction of host and parasite. This can be especially problematic in the case of powdery mildews, such as Erysiphe necator (syn. Uncinula necator), which grow superficially but tightly adhere to the plant epidermis. We developed and refined a simple and efficient technique in which nail polish was used to remove conidia, appressoria, hyphae, conidiophores, and developing ascocarps of E. necator from grapevine (Vitis vinifera) leaves and showed that RNA isolated after removal was not contaminated with V. vinifera RNA. This approach can be applied to expression analyses throughout fungal development and could be extended to other epiphytic pathogens and saprophytes.

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