香蕉穿孔线虫(Radopholus similis)特异性分子检测技术研究  

Species specific molecular diagnosis of burrowing nematode (Radopholus similis)

作　　者：王琼[1] 耿丽凤[1] 张东升[1] 彭德良[1] 

机构地区：[1]中国农业科学院植物保护研究所植物病虫害国家重点实验室,北京100193

出　　处：《植物病理学报》2011年第2期171-177,共7页Acta Phytopathologica Sinica

基　　金：公益性行业(农业)科研专项(200903040)资助;农业部重点实验室开放基金项目(MACKL0905)资助

摘　　要：香蕉穿孔线虫(Radopholus similis)是一种国际公认的极具毁灭性的有害线虫,也是我国重要的进境检疫性有害生物。利用线虫通用引物对香蕉穿孔线虫7个不同群体的ITS进行PCR扩增、克隆及测序,获得ITS片段序列长度为706bp。经与国外香蕉穿孔线虫种群及近缘种的ITS序列进行比对及同源性分析,构建设计了1对香蕉穿孔线虫的特异性引物RsF1/RsR1,特异性扩增片段长度为271 bp;同时引入D2A/D3B作内标,研究出特异性检测香蕉穿孔线虫的一步双重PCR分子技术和方法,该项检测技术特异性好,耗时较短、操作简便、可靠。The burrowing nematode,Radopholus similis is a quarantine nematode pest in the world.Using the universal primer pairs,the ITS region were amplified and sequenced,the ITS length were 706 bp,after analysis and alignment with known ITS sequences of R.similis.One species specific primer pairs,named RsF1/RsR1,was designed to diagnosis of burrowing nematode,the specific fragment was 271 bp.Combined the primer pairs D2A/D3B,the duplex PCR diagnosis for R.similis were developed to distinguish populations of R.similis from other plant parasitic nematode species,only R.similis populations could yield specie speci-fic 271 bp fragment,the other plant parasitic nematodes could not yield species specific fragment.

关 键 词：香蕉穿孔线虫 双重PCR 特异性引物 

分 类 号：S432.45[农业科学—植物病理学]