Kyushu Okinawa Agricultural Research Center, National Agriculture and Food Research Organization, Koshi, Kumamoto 861-1192, Japan.
Hyogo Prefectural Technology Center for Agriculture, Forestry and Fisheries, Kasai, Hyogo 679-0198, Japan.
Plant Dis. 2021 Mar;105(3):643-649. doi: 10.1094/PDIS-05-20-1095-RE. Epub 2021 Jan 19.
An outbreak of downy mildew disease of onion, caused by , in Japan in 2016 necessitated a reevaluation of the primary inoculum sources to optimize disease management. Detection of the pathogen in plants with asymptomatic infection and in soil would guide the application of fungicides according to the extent of infection before disease development. Here, we detected in both plants and soil using newly developed primer sets (Pd ITS and Pd ITS 614) by both conventional and real-time PCR. Validation by real-time PCR with Pd ITS 614 showed that DNA was amplified from symptomless seedlings at 3.7 × 10 to 1.0 × 10 conidium cells/50 mg leaf tissue, suggesting the detection of asymptomatic infection. Real-time PCR with Pd ITS amplified pathogen DNA from field soils at 1.6 × 10 to 8.3 × 10 oospore cells/g of soil. This real-time PCR assay provides a useful tool for identifying and quantifying inoculum sources, which may be the foundation of the design of integrated disease management strategies.
2016 年，日本暴发葱霜霉病，由 引起，这使得人们有必要重新评估主要初侵染源，以优化疾病管理。在发病前，根据感染程度检测无症状感染植物和土壤中的病原菌，可指导杀菌剂的使用。本文中，我们利用新开发的引物（Pd ITS 和 Pd ITS 614），通过常规 PCR 和实时 PCR 检测到了病原菌。利用 Pd ITS 614 进行实时 PCR 验证表明，3.7×10 至 1.0×10 个分生孢子细胞/50mg 叶组织可检测到无症状侵染的病原菌 DNA。Pd ITS 实时 PCR 可从田间土壤中检测到 1.6×10 至 8.3×10 个卵孢子细胞/g 土壤中的病原菌 DNA。该实时 PCR 检测方法为鉴定和定量侵染源提供了一种有用的工具，这可能是综合疾病管理策略设计的基础。
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