甘蔗黑穗病菌DNA提取及PCR检测  

DNA Extraction and PCR Detection of Pathogen of Sugarcane Smut

作　　者：卢文洁[1] 李文凤[1] 黄应昆 罗志明[1] 王明强[1] 王晓燕[1] 

机构地区：[1]云南省农业科学院甘蔗研究所,云南省甘蔗遗传改良重点实验室,云南开远661600

出　　处：《西南农业学报》2012年第2期531-533,共3页Southwest China Journal of Agricultural Sciences

基　　金：公益性行业(农业)科研专项(nyhyzx07-019);现代农业产业技术体系建设专项资金资(nycytx-024-01-09);云南省"人才培养"项目(2008PY087)

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 detected 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.

关 键 词：甘蔗黑穗病菌 基因组DNA 提取方法 PCR检测 

分 类 号：S435.661[农业科学—农业昆虫与害虫防治]