Development of a PCR assay to detect Fusarium poae in wheat
Development of a PCR assay to detect Fusarium poae in wheat
复制标题
开发检测小麦中早熟禾镰孢菌的 PCR 检测方法
DOI：
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发表时间：
1996
期刊：
影响因子：
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通讯作者：
P. Nicholson
中科院分区：
文献类型：
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作者：
D. Parry;P. Nicholson
随机扩增多态性DNA分析进行了一系列分离的F。POAE鉴定所有分离株共有的标记。分离、克隆了两个片段，并将其用于探针DNA的Southern杂交。Poae和来自一系列小麦种子和茎基病原体的分离物。其中一个片段与F.对Poae的部分序列进行了测定，并设计了两对引物（Fp 8 F/R和Fp 82 F/R）用于聚合酶链反应（PCR）。所有分离的F. Poae，而不是来自与谷物穗和种子的疾病相关的任何其它真菌物种。用引物Fp 82 F/R检测F.从被镰刀菌属污染的小麦种子样品提取物中检测到了早熟禾。该系统提供了确定F的存在的可能性。从而避免了通常与疾病的常规诊断和病原体的分离相关的问题。
Random amplified polymorphic DNA assays were carried out on a range of isolates of F. poae to identify markers common to all isolates. Two fragments were isolated, cloned and used to probe Southern blots of DNA from F. poae and isolates from a range of wheat seed and stem base pathogens. One fragment, which hybridized preferentially to DNA of F. poae was partially sequenced and two pairs of primers (Fp8 F/R and Fp82 F/R) were generated for use in the polymerase chain reaction (PCR). Amplification of target DNA occurred following PCR of all isolates of F. poae but not from any of a range of other fungal species associated with diseases of cereal ears and seed. The primer pair Fp82 F/R was used to detect F. poae in extracts from wheat seed samples contaminated with Fusarium species. This system offers the potential to determine the presence of F. poae in wheat and avoid problems commonly associated with conventional diagnosis of the disease and isolation of the pathogen.