{"id":1131,"date":"2013-01-14T16:44:33","date_gmt":"2013-01-14T15:44:33","guid":{"rendered":"http:\/\/sppadbase.ipp.cnr.it\/?p=1131"},"modified":"2013-01-14T16:45:41","modified_gmt":"2013-01-14T15:45:41","slug":"a-new-large-scale-soil-dna-extraction-procedure-and-real-time-pcr-assay-for-the-detection-of-sclerotium-cepivorum-in-soil","status":"publish","type":"post","link":"https:\/\/sppadbase.ipsp.cnr.it\/?p=1131","title":{"rendered":"A new large scale soil DNA extraction procedure and real-time PCR assay for the detection of Sclerotium cepivorum in soil"},"content":{"rendered":"<div class=\"textimage\">\n<p class=\"textimage\"><a href=\"http:\/\/sppadbase.ipp.cnr.it\/?attachment_id=1222\" rel=\"attachment wp-att-1222\"><img decoding=\"async\" loading=\"lazy\" src=\"http:\/\/sppadbase.ipp.cnr.it\/wordpress\/wp-content\/uploads\/2013\/01\/IYRdWGdujO7wja4F1VQWFDl72eJkfbmt4t8yenImKBVaiQDB_Rd1H6kmuBWtceBJ-300x198.jpg\" alt=\"IYRdWGdujO7wja4F1VQWFDl72eJkfbmt4t8yenImKBVaiQDB_Rd1H6kmuBWtceBJ\" width=\"300\" height=\"198\" class=\"alignnone size-medium wp-image-1222\" srcset=\"https:\/\/sppadbase.ipsp.cnr.it\/wp-content\/uploads\/2013\/01\/IYRdWGdujO7wja4F1VQWFDl72eJkfbmt4t8yenImKBVaiQDB_Rd1H6kmuBWtceBJ-300x198.jpg 300w, https:\/\/sppadbase.ipsp.cnr.it\/wp-content\/uploads\/2013\/01\/IYRdWGdujO7wja4F1VQWFDl72eJkfbmt4t8yenImKBVaiQDB_Rd1H6kmuBWtceBJ-150x99.jpg 150w, https:\/\/sppadbase.ipsp.cnr.it\/wp-content\/uploads\/2013\/01\/IYRdWGdujO7wja4F1VQWFDl72eJkfbmt4t8yenImKBVaiQDB_Rd1H6kmuBWtceBJ-400x264.jpg 400w, https:\/\/sppadbase.ipsp.cnr.it\/wp-content\/uploads\/2013\/01\/IYRdWGdujO7wja4F1VQWFDl72eJkfbmt4t8yenImKBVaiQDB_Rd1H6kmuBWtceBJ.jpg 455w\" sizes=\"(max-width: 300px) 100vw, 300px\" \/><\/a>A real-time PCR assay specific for <em>Sclerotium cepivorum<\/em>, the causal agent of white rot in onions, was developed for use with a new DNA extraction method capable of processing up to 1\u00a0kg of soil in weight. The assay was specific for <em>S. cepivorum<\/em> 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\u00b2\u2009=\u20090.89). 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, <em>Botrytis aclada<\/em> and <em>Rhizoctonia solani<\/em> AG8 were also detected, in one soil sample each. <em>Rhizoctonia solani<\/em> AG2-1 was more widespread and was detected in eight different soil samples. The method is therefore suitable to quantify levels of <em>S. cepivorum<\/em> 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.<\/p>\n<div class=\"clearer\"><\/div>\n<p><strong>James W. Woodhall, Kathryn M. Webb, Patricia M. Giltrap, Ian P. Adams, Jeff C. Peters, Giles E. Budge, Neil Boonham<\/strong><br \/>\nEuropean Journal of Plant Pathology<br \/>\nNovember 2012, Volume 134, Issue 3, pp 467-473<\/p>\n<p>DOI: 10.1007\/s10658-012-0025-2<\/p>\n<p>via <a href='http:\/\/link.springer.com\/article\/10.1007\/s10658-012-0025-2\/fulltext.html'>A new large scale soil DNA extraction procedure and real-time PCR assay for the detection of Sclerotium cepivorum in soil<\/a>.<\/p>\n","protected":false},"excerpt":{"rendered":"\n<p class=\"textimage\">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 <span style=\"color:#777\"> . . . &rarr; Read More: <a href=\"https:\/\/sppadbase.ipsp.cnr.it\/?p=1131\">A new large scale soil DNA extraction procedure and real-time PCR assay for the detection of Sclerotium cepivorum in soil<\/a><\/span><\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":[],"categories":[15,3,11,10,86,87],"tags":[23,16,116,58,19,59,152],"_links":{"self":[{"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=\/wp\/v2\/posts\/1131"}],"collection":[{"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=1131"}],"version-history":[{"count":10,"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=\/wp\/v2\/posts\/1131\/revisions"}],"predecessor-version":[{"id":1231,"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=\/wp\/v2\/posts\/1131\/revisions\/1231"}],"wp:attachment":[{"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=1131"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=1131"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=1131"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}