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Development of a real-time PCR assay for detection of Phytophthora kernoviae and comparison of this method with a conventional culturing technique

Lookup NU author(s): Professor Neil Boonham

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Abstract

Phytophthora kernoviae is a recently described pathogen causing leaf blight, aerial dieback and bleeding cankers on trees and shrubs in parts of Great Britain and Ireland and recently reported in New Zealand. This paper describes the development of a TaqMan real-time PCR assay based on internal transcribed spacer (ITS) sequence to aid diagnosis of this pathogen in culture and in plant material. The assay showed no cross reaction with 29 other Phytophthora species, including the closely related species P. boehmeriae, and detected at least 1.2 pg of P. kernoviae DNA per reaction. A rapid and simple method can be used to extract DNA prior to testing by real-time PCR, and a plant internal control assay can be used to aid interpretation of negative results. A comparison of real-time PCR and plating for 526 plant samples collected in the UK indicated that this assay is suitable for use in routine screening for P. kernoviae. © 2011 KNPV.

Publication metadata

Author(s): Hughes KJD, Tomlinson JA, Giltrap PM, Barton V, Hobden E, Boonham N, Lane CR

Publication type: Article

Publication status: Published

Journal: European Journal of Plant Pathology

Year: 2011

Volume: 131

Issue: 4

Pages: 695-703

Print publication date: 01/12/2011

Online publication date: 09/08/2011

ISSN (print): 0929-1873

ISSN (electronic): 1573-8469

Publisher: Springer

URL: https://doi.org/10.1007/s10658-011-9843-x

DOI: 10.1007/s10658-011-9843-x

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