南方镰孢Fusarium meridionale特异性PCR检测方法的建立与应用

Establishment and application of a specific PCR method for Fusarium meridionale detection

为建立快速、稳定的南方镰孢Fusarium meridionale特异性检测方法,对已报道的镰孢属reductase-like基因部分序列进行比对分析,寻找特异性SNP位点,设计出特异性检测引物F-Fm/R-Fm3。利用该引物对包括南方镰孢在内的30株镰孢的基因组DNA进行PCR扩增。结果显示仅在7株南方镰孢中均扩增出400 bp左右的特异性条带。PCR灵敏度试验结果表明该方法的检测灵敏度达到500 pg基因组DNA。

In order to establish a rapid and stable PCR detection method for Fusarium meridionale,the partial reductase-like gene sequences from F.meridionale were analyzed by alignment to screen specific SNPs and a pair of specific primer(F-Fm/R-Fm3)were designed according to the specific SNPs.With the specific primers,the genomic DNAs of 30 strains from 21 Fusarium species,including Fusarium meridionale etc.,were amplified by PCR.The results revealed that a specific amplicon of approximately 400 bp was amplified from seven strains of F.meridionale,while no amplicon was amplified from other 23 control strains and their negative control groups.Moreover,the sensitivity of this method reached 500 pg genomic DNA.