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首页> 外文期刊>Mycologia >Identification of Coniothyrium minitans isolates using PCR amplification of a dispersed repetitive element.
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Identification of Coniothyrium minitans isolates using PCR amplification of a dispersed repetitive element.

机译：使用分散重复元件的PCR扩增来鉴定小锥孢霉菌的分离物。

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摘要

A PCR (polymerase chain reaction) assay was developed to distinguish isolates of the sclerotial mycoparasite Coniothyrium minitans. In particular, the assay uniquely identified isolate A69 which was of interest due to its potential as a biocontrol agent for the plant pathogen Sclerotinia sclerotiorum. Random amplification of polymorphic DNA (RAPD) was used to screen isolates of C. minitans for polymorphic products which distinguished A69 from other C. minitans isolates. A 1.4 kb A69-specific RAPD fragment was cloned and sequenced. The sequence was found to contain 2 tandemly repeated 114 bp units separated by a single base. Southern blot analysis of C. minitans DNA showed that the cloned fragment hybridized to multiple bands in all isolates indicating that it was a dispersed repetitive element. Novel PCR primers were designed from the sequences flanking the tandem repeats. These primers were used to develop a PCR assay which distinguished some isolates of C. minitans and uniquely identified isolate A69. This assay will be used to study the ecology of C. minitans in soil.
机译：开发了一种PCR（聚合酶链反应）测定法，以区分出菌核霉菌寄生小锥虫（Coniothyrium minitans）的分离物。特别地，该测定法独特地鉴定了分离物A69，由于其作为植物病原菌菌核盘菌的生物防治剂的潜力而受到关注。多态性DNA（RAPD）的随机扩增被用来筛选C. minitans分离株的多态性产物，该产物将A69与其他C. minitans分离株区分开。克隆并测序了一个1.4 kb A69特异性的RAPD片段。发现该序列包含2个串联重复的114bp单位，由单个碱基分隔。对微小隐孢子虫DNA的Southern印迹分析表明，克隆的片段与所有分离物中的多条带杂交，表明它是分散的重复元件。从串联重复序列侧翼的序列设计新型PCR引物。这些引物用于开发PCR试验，该试验区分了微小隐孢子虫的某些分离株和独特鉴定的分离株A69。该测定法将用于研究土壤中的微小隐孢子虫的生态学。

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