摘要
叶螨体型微小,田间种群变异较大,传统的形态鉴定方法耗时长,技术难以掌握。为了寻找截形叶螨Tetranychus truncatus Ehara的快速分子鉴定技术,本研究筛选RAPD扩增条带中截形叶螨的特异性引物,并由此转化为稳定的SCAR标记。结果表明,由引物OPB-04扩增截形叶螨出现了一条717 bp的特异性条带(GenBank登录号为JF816665),基于此片段设计了一对特异性的SCAR引物(Tt-303F和Tt-303R),优化PCR扩增条件和程序,从截形叶螨的不同发育阶段中均可成功扩增出一条特异性的303 bp的DNA条带,而在其它近似叶螨种类中未扩增到该条带,且该SCAR引物对在北京地区茄子和菜豆寄主上采集的截形叶螨田间种群上也成功得到了验证。
Spider mites are very tiny animals which frequently display morphological variation between populations. Traditional identification methods take a long time to apply and their key characteristics are difficult to grasp. In order to find a rapid molecular identification method for the spider mite, Tetranychus truncatus Ehara, Random Amplified Polymorphic DNA （RAPD） was used to obtain specific bands and the SCAR marker. One prominent and specific fragment of 717 bp was screened and sequenced （GenBank accession number JF816665） using the OPB-04 primer. A pair of SCAR primers （Tt- 303F and Tt-303R） specific to T. truncatus were designed on the basis of the sequencing results of the 717 bp band. After optimization for the amplification procedure, these primers were used to successfully obtain a 303 bp band from all developmental stages of T. truncatus. This band was not found in other related species. The two SCAR primers were also shown to be effective in monitoring T. truncatus populations collected on eggplant and bean crops in Beijing.
出处
《应用昆虫学报》
CAS
CSCD
北大核心
2013年第2期388-394,共7页
Chinese Journal of Applied Entomology
基金
公益性行业(农业)科研专项(201103020)
