The European and Mediterranean Plant Protection Organization (EPPO) real-time PCR protocol for the detection of ash dieback (Hymenoscyphus fraxineus (T. Kowalski) Barak, Queloz, Hosoya) was optimized for diagnostics on ash (Fraxinus excelsior L.) fruits. Ash fruits were collected from a range of sites in Britain, from trees with various levels of infection. To assess the potential pathway and the presence of H. fraxineus, tissue components of ash fruits as well as ash flowers were tested for the presence of H. fraxineus DNA and RNA. The fungus was detected on ash fruits from trees on all five sampled sites including symptomless, lightly and highly infected sites. DNA of H. fraxineus was detected on the pericarps of all fruit lots and on seeds from fruit lots that included damaged and empty fruits, but not on embryos or flowers. RNA of H. fraxineus was never detected on any of the samples or sample types, indicating that H. fraxineus was only detected in an inactive form (i.e. dormant or dead). The absence of RNA from the pathogen suggests that only spores of H. fraxineus are present on ash fruits. A double hot-water treatment was evaluated as a control measure for the eradication of H. fraxineus on ash fruits; these were treated before and after warm stratification. Large proportions of ash fruits survived the double hot-water treatments, with a 60.4 ± 28.4 per cent germination success rate depending on crop year. Double hot-water treatment of ash fruits proved to be an efficient, low cost and low-tech approach for the eradication of H. fraxineus on ash fruits.

中文翻译：

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从灰 (Fraxinus excelsior) 种子中鉴定、检测和根除 Hymenoscyphus fraxineus

欧洲和地中海植物保护组织 (EPPO) 用于检测灰枯病 (Hymenoscyphus fraxineus (T. Kowalski) Barak, Queloz, Hosoya) 的实时 PCR 方案针对灰烬 (Fraxinus excelsior L.) 果实的诊断进行了优化。从英国的一系列地点，从感染不同程度的树木中收集灰果。为了评估潜在的途径和 H. fraxineus 的存在，测试了白蜡树果实和白蜡树花的组织成分中 H. fraxineus DNA 和 RNA 的存在。在所有五个采样地点的树木灰果上都检测到了真菌，包括无症状、轻度和高度感染的地点。H. fraxineus 的 DNA 在所有水果批次的果皮和包括受损和空果在内的水果批次的种子中检测到，但在胚胎或花上未检测到。H的RNA。从未在任何样品或样品类型中检测到 fraxineus，这表明仅以非活动形式（即休眠或死亡）检测到 H. fraxineus。病原体中 RNA 的缺失表明白蜡树果实上仅存在 H. fraxineus 的孢子。双重热水处理被评估为根除白蜡树上白蜡树的控制措施；这些在温暖分层之前和之后进行了处理。大部分白蜡果在两次热水处理中存活下来，发芽成功率为 60.4 ± 28.4%，具体取决于作物年份。白蜡果双热水处理被证明是一种高效、低成本和低技术的方法来根除白蜡果上的白曲霉。fraxineus 仅以非活动形式（即休眠或死亡）被发现。病原体中 RNA 的缺失表明白蜡树果实上仅存在 H. fraxineus 的孢子。双重热水处理被评估为根除白蜡树上白蜡树的控制措施；这些在温暖分层之前和之后进行了处理。大部分白蜡果在两次热水处理中存活下来，发芽成功率为 60.4 ± 28.4%，具体取决于作物年份。白蜡果双热水处理被证明是一种高效、低成本和低技术的方法来根除白蜡果上的白曲霉。fraxineus 仅以非活动形式（即休眠或死亡）被发现。病原体中 RNA 的缺失表明白蜡树果实上仅存在 H. fraxineus 的孢子。双重热水处理被评估为根除白蜡树上白蜡树的控制措施；这些在温暖分层之前和之后进行了处理。大部分白蜡果在两次热水处理中存活下来，发芽成功率为 60.4 ± 28.4%，具体取决于作物年份。白蜡果双热水处理被证明是一种高效、低成本和低技术的方法来根除白蜡果上的白曲霉。双重热水处理被评估为根除白蜡树上白蜡树的控制措施；这些在温暖分层之前和之后进行了处理。大部分白蜡果在两次热水处理中存活下来，发芽成功率为 60.4 ± 28.4%，具体取决于作物年份。白蜡果双热水处理被证明是一种高效、低成本和低技术的方法来根除白蜡果上的白曲霉。双重热水处理被评估为根除白蜡树上白蜡树的控制措施；这些在温暖分层之前和之后进行了处理。大部分白蜡果在两次热水处理中存活下来，发芽成功率为 60.4 ± 28.4%，具体取决于作物年份。白蜡果双热水处理被证明是一种高效、低成本和低技术的方法来根除白蜡果上的白曲霉。