{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,20]],"date-time":"2026-06-20T00:23:52Z","timestamp":1781915032135,"version":"3.54.5"},"reference-count":38,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2013,7,9]],"date-time":"2013-07-09T00:00:00Z","timestamp":1373328000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["scijournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Pest Management Science"],"published-print":{"date-parts":[[2013,9]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>BACKGROUND<\/jats:title><jats:p><jats:bold>Methyl benzimidazole carbamate (<jats:styled-content style=\"fixed-case\">MBC<\/jats:styled-content>) fungicides are used to control the oilseed rape pathogen <jats:italic>Pyrenopeziza brassicae<\/jats:italic>. Resistance to <jats:styled-content style=\"fixed-case\">MBCs<\/jats:styled-content> has been reported in <jats:italic>P. brassicae<\/jats:italic>, but the molecular mechanism(s) associated with reductions in sensitivity have not been verified in this species. Elucidation of the genetic changes responsible for resistance, hypothesised to be target\u2010site mutations in <jats:italic>\u03b2<\/jats:italic>\u2010tubulin, will enable resistance diagnostics and thereby inform resistance management strategies.<\/jats:bold><\/jats:p><\/jats:sec><jats:sec><jats:title>RESULTS<\/jats:title><jats:p><jats:bold><jats:italic>P. brassicae<\/jats:italic> isolates were classified as sensitive, moderately resistant or resistant to <jats:styled-content style=\"fixed-case\">MBCs<\/jats:styled-content>. Crossing <jats:italic>P. brassicae<\/jats:italic> isolates of different <jats:styled-content style=\"fixed-case\">MBC<\/jats:styled-content> sensitivities indicated that resistance was conferred by a single gene. The <jats:styled-content style=\"fixed-case\">MBC<\/jats:styled-content>\u2010target encoding gene <jats:italic>\u03b2\u2010tubulin<\/jats:italic> was cloned and sequenced. Reduced <jats:styled-content style=\"fixed-case\">MBC<\/jats:styled-content> sensitivity of field isolates correlated with <jats:italic>\u03b2<\/jats:italic>\u2010tubulin amino acid substitutions <jats:styled-content style=\"fixed-case\">L240F<\/jats:styled-content> and <jats:styled-content style=\"fixed-case\">E198A<\/jats:styled-content>. The highest level of <jats:styled-content style=\"fixed-case\">MBC<\/jats:styled-content> resistance was measured for isolates carrying <jats:styled-content style=\"fixed-case\">E198A<\/jats:styled-content>. Negative cross\u2010resistance between <jats:styled-content style=\"fixed-case\">MBCs<\/jats:styled-content> and the fungicides diethofencarb and zoxamide was only measured in <jats:styled-content style=\"fixed-case\">E198A<\/jats:styled-content> isolates. <jats:styled-content style=\"fixed-case\">PCR\u2010RFLP<\/jats:styled-content> was used to screen isolates for the presence of <jats:styled-content style=\"fixed-case\">L240F<\/jats:styled-content> and <jats:styled-content style=\"fixed-case\">E198A<\/jats:styled-content>. The substitutions <jats:styled-content style=\"fixed-case\">E198G<\/jats:styled-content> and <jats:styled-content style=\"fixed-case\">F200Y<\/jats:styled-content> were also detected in <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> samples from <jats:italic>P. brassicae<\/jats:italic> populations after cloning and sequencing of <jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content> products. The frequencies of <jats:styled-content style=\"fixed-case\">L240F<\/jats:styled-content> and <jats:styled-content style=\"fixed-case\">E198A<\/jats:styled-content> in different <jats:italic>P. brassicae<\/jats:italic> populations were quantified by pyrosequencing. There were no differences in the frequencies of these alleles between <jats:italic>P. brassicae<\/jats:italic> populations sampled from different locations or after fungicide treatment regimes.<\/jats:bold><\/jats:p><\/jats:sec><jats:sec><jats:title>CONCLUSIONS<\/jats:title><jats:p><jats:bold>The molecular mechanisms affecting sensitivity to <jats:styled-content style=\"fixed-case\">MBCs<\/jats:styled-content> in <jats:italic>P. brassicae<\/jats:italic> have been identified. Pyrosequencing assays are a powerful tool for quantifying fungicide\u2010resistant alleles in pathogen populations. \u00a9 2013 Society of Chemical Industry<\/jats:bold><\/jats:p><\/jats:sec>","DOI":"10.1002\/ps.3585","type":"journal-article","created":{"date-parts":[[2013,6,11]],"date-time":"2013-06-11T10:11:57Z","timestamp":1370945517000},"page":"1040-1048","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":33,"title":["Detection and molecular characterisation of <i>Pyrenopeziza brassicae<\/i> isolates resistant to methyl benzimidazole carbamates"],"prefix":"10.1002","volume":"69","author":[{"given":"Helen E","family":"Carter","sequence":"first","affiliation":[{"name":"Department of Biological Chemistry and Crop Protection Rothamsted Research Harpenden, Hertfordshire UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hans J","family":"Cools","sequence":"additional","affiliation":[{"name":"Department of 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