剪股颖粒线虫幼虫形态与分子检测方法  

Molecular identification bentgrass nematode, bying the single juvenile.

作　　者：王金成 [1] 马以桂 [1] 周春娜 [2] 杜宇 [3] 黄国明 [1] 丁元明 [3] 谢辉 [2] 

机构地区：[1]天津出入境检验检疫局 3004562 [2]华南农业大学 [3]云南出入境检验检疫局

出　　处：《植物检疫》2005年第2期84-86,共3页Plant Quarantine

基　　金：国家食品安全关键技术专项课题 (2 0 0 1BA80 4A2 2 )

摘　　要：剪股颖粒线虫 (Anguinaagrostis)是牧草、草坪上的重要线虫 ,该线虫在进口草籽中常以幼虫形态出现 ,与其它粒线虫属幼虫形态相似 ,很难快速鉴定到种。本实验通过对单条幼虫DNA进行提取和PCR扩增 ,获得其rDNA的ITS区域 ,测序后与基因库中的剪股颖粒线虫的rDNA的ITS区域序列进行比较 ,其结果与基因库中编码为AF396 343 1的核酸序列吻合 ,从而鉴定为剪股颖粒线虫。此外 ,还使用 5种限制性内切酶对PCR产物进行了酶切 ,获得了剪股颖粒线虫的ITS RFLP指纹图谱。Bentgrass nematode, Anguina agrostis,parasitize the grazing and lawn grass is a kind of dangerous pest. In most cases, only juveniles are isolated from the seed galls infected by Anguina agrostis,so molecular techniques are necessary in the species-level determination. Through PCR amplification using single nematode nucleotide as the template, two internal transcribed spacers, i.e. ITS1and ITS2, were obtained. Sequenced the PCR product and then compared it with the internet-submitted nucleotide sequences accessing through BLASTN in NCBI website. The comparison results confirmed our morphological identification in 2000, which may imply that ITS regions are applicable in the differentiation of the nematode species under genus Anguina.

关 键 词：线虫 草籽 分子检测 基因库 草坪 牧草 幼虫形态 ITS区 DNA PCR产物 

分 类 号：S41[农业科学—植物保护]