{"title":"使用 qPCR 和 LAMP 方法特异性检测 Pyricularia oryzae 病理型 Triticum","authors":"Ken-ichi Ikeda, Kaichi Uchihashi, Itsuki Okuda, Zikai Xiang, Hitoshi Nakayashiki","doi":"10.1007/s10327-023-01162-0","DOIUrl":null,"url":null,"abstract":"<p>Wheat blast is an important disease that affects cross-border food security. The causal pathogen, <i>Pyricularia oryzae</i> (syn. <i>Magnaporthe oryzae</i>) <i>Triticum</i> pathotype (MoT), forms a phylogenetic group distinct from the other <i>P. oryzae</i> pathotypes pathogenic on various gramineous plants. Here we developed MoT-specific primer sets for qPCR and LAMP detection methods, and their amplification specificity and efficiency were tested in comparison with previous reported MoT-specific primers. The amplification efficiency and specificity were affected by the polymerase premix. To evaluate the detection specificity, purified DNA samples of 15 MoT isolates, 89 non-MoT isolates of <i>Pyricularia</i> spp., and crude DNA from wheat plants inoculated with MoT were used. We conclude that the Sp3 primer sets for qPCR and the MoT3<i>L</i> primer sets for LAMP were the best in this research.</p>","PeriodicalId":15825,"journal":{"name":"Journal of General Plant Pathology","volume":"72 1","pages":""},"PeriodicalIF":1.0000,"publicationDate":"2023-12-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Specific detection of Pyricularia oryzae pathotype Triticum using qPCR and LAMP methods\",\"authors\":\"Ken-ichi Ikeda, Kaichi Uchihashi, Itsuki Okuda, Zikai Xiang, Hitoshi Nakayashiki\",\"doi\":\"10.1007/s10327-023-01162-0\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Wheat blast is an important disease that affects cross-border food security. The causal pathogen, <i>Pyricularia oryzae</i> (syn. <i>Magnaporthe oryzae</i>) <i>Triticum</i> pathotype (MoT), forms a phylogenetic group distinct from the other <i>P. oryzae</i> pathotypes pathogenic on various gramineous plants. Here we developed MoT-specific primer sets for qPCR and LAMP detection methods, and their amplification specificity and efficiency were tested in comparison with previous reported MoT-specific primers. The amplification efficiency and specificity were affected by the polymerase premix. To evaluate the detection specificity, purified DNA samples of 15 MoT isolates, 89 non-MoT isolates of <i>Pyricularia</i> spp., and crude DNA from wheat plants inoculated with MoT were used. We conclude that the Sp3 primer sets for qPCR and the MoT3<i>L</i> primer sets for LAMP were the best in this research.</p>\",\"PeriodicalId\":15825,\"journal\":{\"name\":\"Journal of General Plant Pathology\",\"volume\":\"72 1\",\"pages\":\"\"},\"PeriodicalIF\":1.0000,\"publicationDate\":\"2023-12-28\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of General Plant Pathology\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://doi.org/10.1007/s10327-023-01162-0\",\"RegionNum\":4,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"PLANT SCIENCES\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of General Plant Pathology","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.1007/s10327-023-01162-0","RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"PLANT SCIENCES","Score":null,"Total":0}
引用次数: 0
摘要
小麦稻瘟病是一种影响跨境粮食安全的重要病害。病原菌 Pyricularia oryzae (syn. Magnaporthe oryzae) Triticum pathotype (MoT) 与其他在各种禾本科植物上致病的 P. oryzae 病原型形成了一个不同的系统发育群。在此,我们开发了用于 qPCR 和 LAMP 检测方法的 MoT 特异性引物组,并与之前报道的 MoT 特异性引物进行了扩增特异性和效率的比较测试。扩增效率和特异性受聚合酶预混剂的影响。为了评估检测特异性,我们使用了 15 个 MoT 分离物、89 个非 MoT 分离物的 Pyricularia spp.我们得出结论:在本研究中,用于 qPCR 的 Sp3 引物组和用于 LAMP 的 MoT3L 引物组效果最佳。
Specific detection of Pyricularia oryzae pathotype Triticum using qPCR and LAMP methods
Wheat blast is an important disease that affects cross-border food security. The causal pathogen, Pyricularia oryzae (syn. Magnaporthe oryzae) Triticum pathotype (MoT), forms a phylogenetic group distinct from the other P. oryzae pathotypes pathogenic on various gramineous plants. Here we developed MoT-specific primer sets for qPCR and LAMP detection methods, and their amplification specificity and efficiency were tested in comparison with previous reported MoT-specific primers. The amplification efficiency and specificity were affected by the polymerase premix. To evaluate the detection specificity, purified DNA samples of 15 MoT isolates, 89 non-MoT isolates of Pyricularia spp., and crude DNA from wheat plants inoculated with MoT were used. We conclude that the Sp3 primer sets for qPCR and the MoT3L primer sets for LAMP were the best in this research.
期刊介绍:
The Journal of General Plant Pathology welcomes all manuscripts dealing with plant diseases or their control, including pathogen characterization, identification of pathogens, disease physiology and biochemistry, molecular biology, morphology and ultrastructure, genetics, disease transmission, ecology and epidemiology, chemical and biological control, disease assessment, and other topics relevant to plant pathological disorders.