Real-Time PCR for Diagnosing and Quantifying Co-infection by Two Globally Distributed Fungal Pathogens of Wheat
Real-Time PCR for Diagnosing and Quantifying Co-infection by Two Globally Distributed Fungal Pathogens of Wheat
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DOI：
10.3389/fpls.2018.01086
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发表时间：
2018-08-09
影响因子：
5.6
通讯作者：
Zerihun, Ayalsew
中科院分区：
文献类型：
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作者：
Abdullah, Araz S.;Turo, Chala;Zerihun, Ayalsew
共同感染-由多种病原体物种或菌株入侵宿主植物是常见的，并且被认为对病原体生态和进化有影响。尽管合并感染具有明显的重要性，但它们受到的关注有限；部分原因是缺乏适当的定量工具来监测共同感染的病原体。在这里，我们报告了一项双工实时PCR检测，该检测同时区分和量化了小麦两种全球重要的真菌病原体：小麦Pyrenophora tritrii -repentis和nodorum Parastagonospora nodorum的共同感染。这些真菌具有共同的特征和宿主物种，对传统的疾病诊断和随后的管理策略提出了挑战。该试验使用唯一指定的荧光探针来量化真菌生物量作为核酸当量。该探针提供高度特异性的目标定量，对非目标密切相关的真菌物种和宿主基因进行准确的区分。真菌靶点的定量在宽范围内呈线性（5000-0.5 pg DNA μ l(-1)），重现性高（RSD）
Co-infections - invasions of a host-plant by multiple pathogen species or strains are common, and are thought to have consequences for pathogen ecology and evolution. Despite their apparent significance, co-infections have received limited attention; in part due to lack of suitable quantitative tools for monitoring of co-infecting pathogens. Here, we report on a duplex real-time PCR assay that simultaneously distinguishes and quantifies co-infections by two globally important fungal pathogens of wheat: Pyrenophora tritici-repentis and Parastagonospora nodorum. These fungi share common characteristics and host species, creating a challenge for conventional disease diagnosis and subsequent management strategies. The assay uses uniquely assigned fluorogenic probes to quantify fungal biomass as nucleic acid equivalents. The probes provide highly specific target quantification with accurate discrimination against non-target closely related fungal species and host genes. Quantification of the fungal targets is linear over a wide range (5000-0.5 pg DNA mu l(-1)) with high reproducibility (RSD