摘要
通过对已测序和已报道的葡萄霜霉病菌[Plasmopara viticola(Berket Curtis)Ber.et de Toni]细胞色素c氧化酶亚型2(cytochrome c oxidaseⅡ,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FPv/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600bp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3.3pg/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%。
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.
出处
《植物保护》
CAS
CSCD
北大核心
2014年第2期99-102,108,共5页
Plant Protection
基金
公益性行业(农业)科研专项(201203035)
国家现代农业产业技术体系专项基金(nycytx-30)
