Adam Kuzdraliński, Hubert Szczerba, Anna Kot, Agnieszka Ostrowska, Michał Nowak, Marta Muszyńska
{"title":"小麦蓝灰菌PCR检测新方法的建立与应用。","authors":"Adam Kuzdraliński, Hubert Szczerba, Anna Kot, Agnieszka Ostrowska, Michał Nowak, Marta Muszyńska","doi":"10.1159/000494432","DOIUrl":null,"url":null,"abstract":"<p><p>We developed new PCR assays that target beta-tubulin (TUB2) and 14 alpha-demethylase (CYP51) genes and used them for the species-specific detection of Blumeria graminis f. sp. tritici (Bgt). Based on fungi DNA sequences available in the NCBI (National Center for Biotechnology Information) GenBank database we developed simplex and duplex PCR assays. The specificities of the primer sets were evaluated using environmental samples of wheat leaves collected during the 2015/2016 growing season across Poland. Primer sets LidBg17/18 and LidBg21/22 strongly amplified fragments of the expected length for all 67 tested samples. Primer specificity was confirmed using field samples of Zymoseptoria tri-tici, Puccinia triticina (syn. P. recondita f. sp. tritici), P. striiformis f. sp. tritici, and Pyrenophora tritici-repentis.</p>","PeriodicalId":16370,"journal":{"name":"Journal of Molecular Microbiology and Biotechnology","volume":"28 3","pages":"137-146"},"PeriodicalIF":1.2000,"publicationDate":"2018-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1159/000494432","citationCount":"2","resultStr":"{\"title\":\"Development and Application of a New PCR Method for Detection of Blumeria graminis f. sp. tritici.\",\"authors\":\"Adam Kuzdraliński, Hubert Szczerba, Anna Kot, Agnieszka Ostrowska, Michał Nowak, Marta Muszyńska\",\"doi\":\"10.1159/000494432\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>We developed new PCR assays that target beta-tubulin (TUB2) and 14 alpha-demethylase (CYP51) genes and used them for the species-specific detection of Blumeria graminis f. sp. tritici (Bgt). Based on fungi DNA sequences available in the NCBI (National Center for Biotechnology Information) GenBank database we developed simplex and duplex PCR assays. The specificities of the primer sets were evaluated using environmental samples of wheat leaves collected during the 2015/2016 growing season across Poland. Primer sets LidBg17/18 and LidBg21/22 strongly amplified fragments of the expected length for all 67 tested samples. Primer specificity was confirmed using field samples of Zymoseptoria tri-tici, Puccinia triticina (syn. P. recondita f. sp. tritici), P. striiformis f. sp. tritici, and Pyrenophora tritici-repentis.</p>\",\"PeriodicalId\":16370,\"journal\":{\"name\":\"Journal of Molecular Microbiology and Biotechnology\",\"volume\":\"28 3\",\"pages\":\"137-146\"},\"PeriodicalIF\":1.2000,\"publicationDate\":\"2018-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1159/000494432\",\"citationCount\":\"2\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Molecular Microbiology and Biotechnology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1159/000494432\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2018/12/6 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q2\",\"JCRName\":\"Biochemistry, Genetics and Molecular Biology\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Molecular Microbiology and Biotechnology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1159/000494432","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2018/12/6 0:00:00","PubModel":"Epub","JCR":"Q2","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 2
摘要
建立了针对β -微管蛋白(TUB2)和14 α -去甲基化酶(CYP51)基因的PCR检测方法,并将其用于小麦蓝灰菌(Blumeria graminis f. sp. tritici, Bgt)的种特异性检测。基于NCBI (National Center for Biotechnology Information) GenBank数据库中的真菌DNA序列,我们开发了单链和双链PCR检测方法。利用波兰2015/2016生长季收集的小麦叶片环境样本,对引物组的特异性进行了评估。引物LidBg17/18和LidBg21/22对所有67个测试样品的预期长度进行了强烈扩增。利用田间样品对稻瘟病菌(Zymoseptoria tritici)、小麦锈病菌(p.r recondita f.p tritici)、小麦纹状菌(p.s striiformis f.p tritrireentis)和稻瘟病菌(Pyrenophora tritrii -repentis)的引物进行特异性鉴定。
Development and Application of a New PCR Method for Detection of Blumeria graminis f. sp. tritici.
We developed new PCR assays that target beta-tubulin (TUB2) and 14 alpha-demethylase (CYP51) genes and used them for the species-specific detection of Blumeria graminis f. sp. tritici (Bgt). Based on fungi DNA sequences available in the NCBI (National Center for Biotechnology Information) GenBank database we developed simplex and duplex PCR assays. The specificities of the primer sets were evaluated using environmental samples of wheat leaves collected during the 2015/2016 growing season across Poland. Primer sets LidBg17/18 and LidBg21/22 strongly amplified fragments of the expected length for all 67 tested samples. Primer specificity was confirmed using field samples of Zymoseptoria tri-tici, Puccinia triticina (syn. P. recondita f. sp. tritici), P. striiformis f. sp. tritici, and Pyrenophora tritici-repentis.
期刊介绍:
We are entering a new and exciting era of microbiological study and application. Recent advances in the now established disciplines of genomics, proteomics and bioinformatics, together with extensive cooperation between academic and industrial concerns have brought about an integration of basic and applied microbiology as never before.