Differentiation and quantification of the cereal eyespot fungi Tapesia yallundae and Tapesia acuformis using a PCR assay
Differentiation and quantification of the cereal eyespot fungi Tapesia yallundae and Tapesia acuformis using a PCR assay
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DOI：
10.1046/j.1365-3059.1997.d01-74.x
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发表时间：
1997-12-01
期刊：
影响因子：
2.7
通讯作者：
Joyce, D
中科院分区：
文献类型：
--
作者：
Nicholson, P;Rezanoor, HN;Joyce, D
对黄带绦虫和尖锐带绦虫的分离株进行了随机扩增多态DNA分析。对两个物种的分离物共同的扩增产物进行克隆，并从每个序列中产生用于常规聚合酶链式反应的引物。这对来自黄粉病特异的RAPD标记的引物只能从黄粉病分离物的DNA中扩增出产物，而不能从与谷类茎基病相关的一系列其他真菌物种的DNA中扩增出产物。类似地，从尖端弓形虫特异的RAPD标记产生的引物对仅从尖端锥虫分离株的DNA中扩增出产物。利用这两对引物建立了两种带状线虫的定量分析方法，利用竞争性PCR方法测定了每种带状线虫在温室和田间接种的谷类寄主上的定植水平，并与常规苗期病害评估结果进行了比较。
Isolates of Tapesia yallundae and Tapesia acuformis were subjected to Random Amplified Polymorphic DNA (RAPD) assay. Amplification products common to isolates of either species were cloned and primers were generated from each sequence for use in conventional PCR. The primer pair derived from a T. yallundae specific RAPD marker amplified a product only from DNA of T. yallundae isolates and not from DNA of a range of other fungal species associated with the stem base disease complex of cereals. Similarly, the primer pair generated from a T. acuformis-specific RAPD marker amplified a product only from DNA of T. acuformis isolates. Quantitative assays were developed for both species of Tapesia from these primer pairs, using competitive PCR Competitive PCR was used to determine the level of colonization of seedlings by each species in glasshouse- and field-inoculated cereal hosts and results compared to those for conventional seedling disease assessment.