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

Content Provider	Springer Nature Link
Author	Woodhall, James W. Webb, Kathryn M. Giltrap, Patricia M. Adams, Ian P. Peters, Jeff C. Budge, Giles E. Boonham, Neil
Copyright Year	2012
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 (R² = 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.
Starting Page	467
Ending Page	473
Page Count	7
File Format	PDF
ISSN	09291873
Journal	European Journal of Plant Pathology
Volume Number	134
Issue Number	3
e-ISSN	15738469
Language	English
Publisher	Springer Netherlands
Publisher Date	2012-07-14
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Onion White rot TaqMan Allium Rhizoctonia solani Botrytis Agriculture Ecology Plant Pathology Life Sciences Plant Sciences
Content Type	Text
Resource Type	Article
Subject	Plant Science Horticulture Agronomy and Crop Science

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