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首页> 外文期刊>Journal of general plant pathology >DNA fingerprinting of Pyricularia grisea by rep-PCR using a single primer based on the terminal inverted repeat from either of the transposable elements Pot2 and MGR586
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DNA fingerprinting of Pyricularia grisea by rep-PCR using a single primer based on the terminal inverted repeat from either of the transposable elements Pot2 and MGR586

机译：使用基于转座子Pot2和MGR586中任一末端末端反向重复序列的单一引物通过rep-PCR进行稻瘟病菌的DNA指纹识别

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摘要

We investigated the use of single primers complementary to sequences in the terminal inverted repeat (TIR) of either Pot2 or MGR586, transposable elements found in Pyricularia grisea, for DNA fingerprinting by repetitive-element-based polymerase chainreaction (rep-PCR). Under standard amplification conditions, rep-PCR with each single primer generated distinct fingerprint patterns among rice-infecting P. grisea isolates collected in Japan. With the Pot2-TIR primer, bands ranging in size from 0.2 to8kb and in number from 8 to 13 per isolate were amplified. Although fewer bands were amplified with the MGR586-TIR primer, this molecular technique should be more reliable to identify and classify P. grisea isolates by combining the data of fingerprint patterns from each TIR primer. In a cluster analysis based on DNA fingerprints from this rep-PCR with the Pot2-TIR primer, 10 reference isolates and 12 field isolates from Saga Prefecture in 2002 were separated into six clonal lineages. We also demonstrated that the 12 field isolates belonged to one clonal lineage. Thus, this rep-PCR method using the single primer Pot2-TIR will be useful for the analysis of the population structure of rice blast pathogens.
机译：我们研究了使用互补于Pot2或MGR586（在稻瘟病菌中的转座子）的末端反向重复序列（TIR）中的序列的单个引物，通过基于重复元素的聚合酶链反应（rep-PCR）进行DNA指纹识别。在标准扩增条件下，在日本收集的水稻感染稻瘟病菌中，每个引物的rep-PCR产生不同的指纹图谱。使用Pot2-TIR引物，每个分离物的条带大小在0.2到8kb之间，条带从8到13条之间。尽管使用MGR586-TIR引物扩增的条带较少，但该分子技术应更可靠地通过结合每个TIR引物的指纹图谱数据来鉴定和分类稻瘟病菌。在使用Pot2-TIR引物基于基于rep-PCR的DNA指纹的聚类分析中，将2002年佐贺县的10个参考分离株和12个野外分离株分为6个克隆谱系。我们还证明了这12个野外分离株属于一个克隆谱系。因此，这种使用单引物Pot2-TIR的rep-PCR方法将对稻瘟病菌的种群结构分析有用。

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