Genomics-Based Marker Discovery and Diagnostic Assay Development for Wheat Blast
Genomics-Based Marker Discovery and Diagnostic Assay Development for Wheat Blast
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DOI：
10.1094/pdis-04-16-0500-re
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发表时间：
2017-01-01
期刊：
影响因子：
4.5
通讯作者：
Pedley, Kerry F.
中科院分区：
文献类型：
--
作者：
Pieck, Michael L.;Ruck, Amy;Pedley, Kerry F.
麦瘟已成为南美洲小麦生产的主要威胁。尽管最初仅限于巴西，但此后在阿根廷、玻利维亚和巴拉圭等邻国也观察到了这种疾病，最近从孟加拉国受感染的小麦中分离出了病原体，即 Magnaporthe oryzae Triticum 致病型。人们越来越担心这种病原体可能会继续传播到世界其他地区，包括美国，几种米霉致病型在美国流行。米霉致病型在形态上无法区分，因此必须进行基因型表征。小麦瘟病的症状包括头部白化，这与镰刀菌赤霉病的症状非常相似，这使得监测田间病原体存在的工作进一步复杂化。我们使用基于基因组学的方法来鉴定米霉小麦致病型特有的分子标记。其中一个标记 MoT3 被选择用于开发基于聚合酶链式反应 (PCR) 的诊断测定，使用从多种宿主物种中收集的 284 个米霉分离株的 DNA 评估其特异性。传统的 PCR 引物设计用于扩增 361 bp 的产物，并且该方案可以从低至 0.1 ng 的纯化 DNA 中进行一致的扩增。还使用镰刀菌属评估了基于 MoT3 的测定的特异性。 DNA，未检测到扩增子。
Wheat blast has emerged as a major threat to wheat production in South America. Although originally restricted to Brazil, the disease has since been observed in the neighboring countries of Argentina, Bolivia, and Paraguay and recently the pathogen, Magnaporthe oryzae Triticum pathotype, was isolated from infected wheat in Bangladesh. There is growing concern that the pathogen may continue to spread to other parts of the world, including the United States, where several M. oryzae pathotypes are endemic. M. oryzae pathotypes are morphologically indistinguishable and, therefore, must be characterized genotypically. Symptoms of wheat blast include bleaching of the head, which closely resembles the symptoms of Fusarium head blight, further complicating efforts to monitor for the presence of the pathogen in the field. We used a genomics-based approach to identify molecular markers unique to the Triticum pathotype of M. oryzae. One of these markers, MoT3, was selected for the development of a polymerase chain reaction (PCR)-based diagnostic assay that was evaluated for specificity using DNA from 284 M. oryzae isolates collected from a diverse array of host species. Conventional PCR primers were designed to amplify a 361-bp product, and the protocol consistently amplified from as little as 0.1 ng of purified DNA. The specificity of the MoT3-based assay was also evaluated using Fusarium spp. DNA, from which no amplicons were detected.