{"id":1129,"date":"2013-01-15T16:14:18","date_gmt":"2013-01-15T15:14:18","guid":{"rendered":"http:\/\/sppadbase.ipp.cnr.it\/?p=1129"},"modified":"2013-01-15T17:02:21","modified_gmt":"2013-01-15T16:02:21","slug":"the-development-of-a-validated-real-time-taqman-pcr-for-detection-of-stagonosporopsis-andigena-and-s-crystalliniformis-in-infected-leaves-of-potato-and-tomato","status":"publish","type":"post","link":"https:\/\/sppadbase.ipsp.cnr.it\/?p=1129","title":{"rendered":"The development of a validated real-time (TaqMan) PCR for detection of Stagonosporopsis andigena and S. crystalliniformis in infected leaves of potato and tomato"},"content":{"rendered":"
\n

\"HdZ2LhspaThlWr3J1r4Vzjl72eJkfbmt4t8yenImKBVaiQDB_Rd1H6kmuBWtceBJ\"<\/a>Stagonosporopsis andigena<\/em> and S. crystalliniformis<\/em> are serious foliage pathogens on potato (Solanum tuberosum)<\/em> and tomato (Solanum lycopersicum<\/em>). As both species have been recorded only in the Andes area, S. andigena<\/em> is listed as an A1 quarantine organism in Europe. The actin region of isolates of Stagonosporopsis<\/em> and allied species of Boeremia<\/em>, Didymella<\/em>, Peyronellaea<\/em> and Phoma<\/em> was amplified using generic primers. DNA sequence differences of the actin gene were utilised to develop species-specific real-time (TaqMan) PCR assays for the detection of S. andigena<\/em> and S. crystalliniformis<\/em> in leaves of potato or tomato. The specificity of the TaqMan PCR assays was determined on genomic DNA extracted from two S. andigena<\/em> and two S. crystalliniformis<\/em> isolates and 16 selected isolates of Stagonosporopsis<\/em>, Phoma<\/em> and Boeremia<\/em>, which are the closest relatives. The validation of the methods developed included the DNA extraction and the TaqMan PCR assays. The performance criteria specificity, analytical sensitivity, reproducibility, repeatability and robustness of the TaqMan PCR assays demonstrated the reliability of both methods for the detection of S. andigena<\/em> and S. crystalliniformis<\/em> in leaf material. The TaqMan PCR assays were tested on symptomatic leaves of potato and tomato that were obtained after artificial inoculation of detached leaves with both pathogens under quarantine conditions. In the artificial inoculation experiments both S. andigena<\/em> and S. crystalliniformis<\/em> caused leaf infections on potato and tomato.<\/p>\n

<\/div>\n

Johannes de Gruyter, Marga P. E. van Gent-Pelzer, Joyce H. C. Woudenberg, Patricia C. J. van Rijswick, Ellis T. M. Meekes, Pedro W. Crous, Peter J. M. Bonants<\/strong>
\nEuropean Journal of Plant Pathology
\nOctober 2012, Volume 134, Issue 2, pp 301-313<\/p>\n

DOI: 10.1007\/s10658-012-9990-8<\/p>\n

via The development of a validated real-time (TaqMan) PCR for detection of Stagonosporopsis andigena and S. crystalliniformis in infected leaves of potato and tomato – Springer<\/a>.<\/p>\n<\/div>\n","protected":false},"excerpt":{"rendered":"\n

Stagonosporopsis andigena and S. crystalliniformis are serious foliage pathogens on potato (Solanum tuberosum) and tomato (Solanum lycopersicum). As both species have been recorded only in the Andes area, S. andigena is listed as an A1 quarantine organism in Europe. The actin region of isolates of Stagonosporopsis and allied species of Boeremia, Didymella, Peyronellaea . . . → Read More: The development of a validated real-time (TaqMan) PCR for detection of Stagonosporopsis andigena and S. crystalliniformis in infected leaves of potato and tomato<\/a><\/span><\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":[],"categories":[15,3,11,92,93,94],"tags":[23,16,116,58,19,59,156,157],"_links":{"self":[{"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=\/wp\/v2\/posts\/1129"}],"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=1129"}],"version-history":[{"count":13,"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=\/wp\/v2\/posts\/1129\/revisions"}],"predecessor-version":[{"id":1282,"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=\/wp\/v2\/posts\/1129\/revisions\/1282"}],"wp:attachment":[{"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=1129"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=1129"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=1129"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}