{"id":1134,"date":"2013-01-15T17:30:39","date_gmt":"2013-01-15T16:30:39","guid":{"rendered":"http:\/\/sppadbase.ipp.cnr.it\/?p=1134"},"modified":"2013-01-15T17:36:47","modified_gmt":"2013-01-15T16:36:47","slug":"real-time-pcr-as-a-promising-tool-to-monitor-growth-of-venturia-spp-in-scab-susceptible-and-resistant-apple-leaves","status":"publish","type":"post","link":"https:\/\/sppadbase.ipsp.cnr.it\/?p=1134","title":{"rendered":"Real-time PCR as a promising tool to monitor growth of Venturia spp. in scab-susceptible and -resistant apple leaves"},"content":{"rendered":"
\n

\"nM2EmmPPZPjQhzkRtf9i3Tl72eJkfbmt4t8yenImKBVaiQDB_Rd1H6kmuBWtceBJ\"<\/a>Apple scab, the most important disease of apple worldwide, is caused by Venturia inaequalis<\/em>. Currently, evaluation of fungal pathogenicity and host resistance is based on a symptomatic disease rating. However, this method does not provide an accurate measurement of the degree of infection and cannot detect early fungal development in symptomless leaves. In this study, a Venturia<\/em>-specific real-time PCR assay was developed using primers designed around the specific internal transcribed spacer 2 (ITS2) region of the 5.8S rRNA gene. Using SYBR\u00ae Green I technology, the assay can accurately quantify Venturia<\/em> DNA over a concentration range of at least five orders of magnitude. Detection sensitivities were in the order of 100\u00a0fg. The method was used to quantify Venturia<\/em> genomic DNA levels in leaves of three apple cultivars with different levels and types of scab resistance and artificially infected with V. inaequalis<\/em>. The assay clearly discriminated between Venturia<\/em> levels in monogenic resistant (\u2018Topaz\u2019), polygenic resistant (\u2018Discovery\u2019), and susceptible (\u2018Golden Delicious\u2019) cultivars, and proved especially useful to quantify pathogen levels during the initial latent stage of infection. The real-time PCR data of \u2018Golden Delicious\u2019 were consistent with the observed evolution of the degree of sporulation during a time-course experiment. Although measurements were influenced by the presence of co-extracted PCR-inhibitors, the impact of these compounds was independent of the apple cultivar or the initial amount of fungal DNA present. In conclusion, real-time PCR amplification of the ITS2-5.8S rDNA of Venturia<\/em> spp. is a faster, more objective and more sensitive method to monitor fungal growth and to evaluate host resistance than phenotypic disease rating scores.<\/p>\n

<\/div>\n

Bruno Dani\u00ebls, Anke De landtsheer, Rozemarijn Dreesen, Mark W. Davey, Johan Keulemans<\/strong>
\nEuropean Journal of Plant Pathology
\nDecember 2012, Volume 134, Issue 4, pp 821-833<\/p>\n

DOI:10.1007\/s10658-012-0058-6<\/p>\n

via Real-time PCR as a promising tool to monitor growth of Venturia spp. in scab-susceptible and -resistant apple leaves – Springer<\/a>.<\/p>\n","protected":false},"excerpt":{"rendered":"\n

Apple scab, the most important disease of apple worldwide, is caused by Venturia inaequalis. Currently, evaluation of fungal pathogenicity and host resistance is based on a symptomatic disease rating. However, this method does not provide an accurate measurement of the degree of infection and cannot detect early fungal development in symptomless leaves. In . . . → Read More: Real-time PCR as a promising tool to monitor growth of Venturia spp. in scab-susceptible and -resistant apple leaves<\/a><\/span><\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":[],"categories":[15,3,11,10,95,96],"tags":[23,16,116,58,19,59,158],"_links":{"self":[{"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=\/wp\/v2\/posts\/1134"}],"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=1134"}],"version-history":[{"count":12,"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=\/wp\/v2\/posts\/1134\/revisions"}],"predecessor-version":[{"id":1290,"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=\/wp\/v2\/posts\/1134\/revisions\/1290"}],"wp:attachment":[{"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=1134"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=1134"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=1134"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}