甘蔗黑穗病菌DNA提取及PCR检测

摘要

本研究利用尿素混合液直接从甘蔗黑穗病菌孢子提取基因组DNA进行PCR检测鉴定,经过反复试验,建立了一种简便、快速的检测鉴定方法.利用建立的方法对12个甘蔗黑穗病样品进行病菌基因组DNA提取,将甘蔗黑穗病菌特异性引物进行PCR检测,均扩增出大小为420 bp的预期DNA片段条带.PCR扩增产物经克隆测序,与GenBank数据库中的甘蔗黑穗病病原菌序列比对,同源性达99％.%The genomic DNA were extracted directly from sugarcane smut spores for PCR detection by using urea mixture in this study. After test again and again, A simple and rapid method of detection an identification for pathogen PCR was established. The expected bands of DNA fragment with 420 bp were amplified after extracting the genomic DNA of 12 samples from different planting sugarcane areas in this method and delected by PCR with specific primers of sugarcane smut. The homology of sequence of PCR amplified products was 99 % compared with the sequence of sugarcane smut pathogen in the GenBank after cloning and sequencing PCR amplified products.