腐烂茎线虫种内群体特异性检测研究  

Species-specific detection of inter-populations of Ditylenchus destructor

作　　者：刘斌[1] 梅圆圆[1] 郑经武[1] 

机构地区：[1]浙江大学生物技术研究所,浙江杭州310029

出　　处：《浙江大学学报（农业与生命科学版）》2007年第5期490-496,共7页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：杭州市科技局资助项目(200432239).

摘　　要：对腐烂茎线虫（Ditylenchus destructor Thorne）7个不同地理群体rDNA中的ITS区进行PCR-RFLP和序列测定,发现种内群体间无论在序列长度还是酶切图谱均存在一定的变异.根据其序列特征设计出种特异性引物,对所供试的腐烂茎线虫群体及属间不同种群体样品的rDNAITS区相应区段扩增以验证其特异性,同时对分别由1～5条腐烂茎线虫成虫和幼虫提取的DNA进行特异性扩增以验证其灵敏度.结果表明：所有供试的腐烂茎线虫群体均可以特异性扩增一个346bp的片段,而所有供试的属外种均没有扩增片段;由1～5条腐烂茎线虫成虫和4～5条幼虫进行的特异性扩增也获得稳定的目的片段.显示该特异性引物具有较高的稳定性、特异性和灵敏度,能快速、准确地检测出腐烂茎线虫.　PCR-RFLP of rDNA ITS regions of seven geographic populations of D.destructor were made,and the variation in the sequence length and digestion patterns among intra-species populations was detected.According to the sequence character of the ITS region of rDNA of D.destructor,a pair of specific primers were designed,and effectiveness was proved by specific amplifying the corresponding ITS region of all the D.destructor populations.To test the sensitivity of the primers,one to five adult and juvenile specimens of D.destructor were chosen for specific amplification.All the D.destructor populations yielded a specific fragment with the length of 346 bp,and the inter-genus species had no specific amplification.The target fragment was also amplified with 1 to 5 adult specimens and 4 to 5 larva specimens of D.destructor.It is indicated that the primers designed for specific detection of D.destructor were steady,specific and sensitive,and the procedure was fast.

关 键 词：腐烂茎线虫 rDNAITS区 特异性检测 灵敏性 

分 类 号：Q78[生物学—分子生物学] S435.313[农业科学—农业昆虫与害虫防治]