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Development and application of a TaqMan real-time PCR assay for rapid detection of Magnaporthe poae

机译：TaqMan实时荧光定量PCR检测试剂盒用于快速检测Magnaporthe poae的开发和应用

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摘要

In North America, one of the most important root diseases of Poa and Festuca turf is summer patch, caused by Magnaporthe poae. Detection and identification of M. poae in infected roots by conventional culture-based methods is difficult and time consuming, typically taking 3 wk or longer to accomplish. In this study, a culture-independent, TaqMan real-time PCR assay was developed for the detection of M. poae from the roots of fungicidetreated and non-treated Kentucky bluegrass (Poa pratensis) turf. The assay was validated with the target pathogen, closely related fungal species and a number of other microorganisms that inhabit the same host and soil environment. This assay was more sensitive (could detect as little as 3.88 pg genomic DNA of M. poae), rapid and accurate compared to direct microscopic observation and isolation on a selective medium. The real-time PCR detection results corresponded closely to visual assessments of disease severity in the field. Utilization of this assay in diagnostic laboratories will enable turfgrass managers to more quickly and effectively detect and potentially reduce fungicide usage through early and accurate identification of the pathogen.
机译：在北美，Poa和Festuca草皮最重要的根系疾病之一是夏季斑块，由Magnaporthe poae引起。通过传统的基于培养的方法检测和鉴定感染根中的M. poae既困难又耗时，通常需要3周或更长的时间才能完成。在这项研究中，开发了一种不依赖培养物的TaqMan实时PCR检测试剂盒，用于从经过杀真菌剂处理和未处理的肯塔基州蓝草（Poa pratensis）草皮的根中检测毛孢霉。使用目标病原体，密切相关的真菌物种以及居住在相同宿主和土壤环境中的许多其他微生物对试验进行了验证。与直接显微镜观察和在选择性培养基上分离相比，该测定法更加灵敏（可以检测到3.88 pg的波分枝杆菌基因组DNA），快速，准确。实时PCR检测结果与现场疾病严重程度的视觉评估非常接近。通过在诊断实验室中使用该检测方法，草皮草管理者可以通过早期和准确地识别病原体，更快，更有效地检测并可能减少杀菌剂的使用。

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