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A new large scale soil DNA extraction procedure and real-time PCR assay for the detection of Sclerotium cepivorum in soil

Lookup NU author(s): Professor Giles Budge, Professor Neil Boonham

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Abstract

A real-time PCR assay specific for Sclerotium cepivorum, the causal agent of white rot in onions, was developed for use with a new DNA extraction method capable of processing up to 1 kg of soil in weight. The assay was specific for S. cepivorum when tested against 24 isolates representative of 14 closely related species and other pathogens of onion. The assay was highly sensitive when used with soil DNA extracted using the new DNA extraction procedure. In three different field soils tested, a good relationship between cycle threshold (Ct) and number of sclerotia was observed (R 2 = 0. 89). Twenty-nine soil samples from onion and leek crops were obtained and the pathogen was detected in four samples. All four positive samples were associated with current or past outbreaks of white rot of onion. Additional assays were also used on the 29 field soil samples, Botrytis aclada and Rhizoctonia solani AG8 were also detected, in one soil sample each. Rhizoctonia solani AG2-1 was more widespread and was detected in eight different soil samples. The method is therefore suitable to quantify levels of S. cepivorum in soil samples, with the added advantage that the technique allows other soil pathogens of interest to be assayed from the same DNA sample. The bulk soil DNA extraction method described here has the potential to be used to detect soil-borne pests and pathogens for other crops in a wide range of soil types. © 2012 Crown Copyright 2012.


Publication metadata

Author(s): Woodhall JW, Webb KM, Giltrap PM, Adams IP, Peters JC, Budge GE, Boonham N

Publication type: Article

Publication status: Published

Journal: European Journal of Plant Pathology

Year: 2012

Volume: 134

Issue: 3

Pages: 467-473

Print publication date: 01/11/2012

Online publication date: 14/07/2012

ISSN (print): 0929-1873

ISSN (electronic): 1573-8469

Publisher: Springer

URL: https://doi.org/10.1007/s10658-012-0025-2

DOI: 10.1007/s10658-012-0025-2


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