| 秦文韬，黄晓庆，孔繁芳，王忠跃*，张昊*.葡萄霜霉病菌PCR检测方法的建立与应用[J].植物保护,2014,40(2):99-102. |
| 葡萄霜霉病菌PCR检测方法的建立与应用 |
| Establishment and application of PCR for detection of Plasmopara viticola |
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| DOI： |
| 中文关键词: 葡萄霜霉病菌 cox2 聚合酶链式反应 检测 |
| 英文关键词:Plasmopara viticola cox2 PCR detection |
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| 中文摘要: |
| 通过对已测序和已报道的葡萄霜霉病菌［Plasmopara viticola (Berk et Curtis) Ber.et de Toni］细胞色素c氧化酶亚型2（cytochrome c oxidase Ⅱ，cox2）的基因序列进行同源性比对分析，设计合成了一对用于P.viticola的检测引物F Pv/R Pv。利用该引物对包括P.viticola在内的30种常见植物病原真菌（包括15种Plasmopara属真菌、8种葡萄常见致病菌）的基因组DNA进行了PCR扩增，验证引物F Pv/R Pv的特异性，结果表明：该引物特异性强，仅P.viticola基因组DNA作为模板的PCR扩增产物呈现一条600 bp左右的特异性条带，其他参照菌株及阴性对照均无任何条带；灵敏度验证结果表明，该检测法可以检测出3.3 pg/μL 水平的P.viticola基因组DNA；应用该方法对来自全国不同葡萄产区的不同品种的78份葡萄霜霉病菌进行了检测，结果表明，该检测法适用范围广，且检测准确率达100%。 |
| 英文摘要: |
| This study designed a pair of primers F Pv/R Pv for Plasmopara viticola detection based on the differences among sequenced and reported cox2 gene sequences of P.viticola. Totally 30 kinds of plant pathogens, including 15 kinds of fungi belonging to Plasmopara and 8 kinds of grape pathogens, were screened to validate the primers F Pv/R Pv. The results showed that the primers F Pv/R Pv were of high specificity, and only a single product of about 600 bp was amplified from P.viticola genomic DNA. The sensitivity of the detection was 3.3 pg/μL genomic DNA per 20 μL PCR reaction volume. Genomic DNAs of 78 samples from different grape plantation areas in China were specifically detected by PCR assay with F Pv/R Pv, suggesting that this method was applicable to a wide scope with 100% detection accuracy rate. |
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