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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 body size is small, large variation in field populations, the traditional morphological identification method takes a long time, technology is difficult to grasp. In order to search for the rapid molecular identification technique of Tetranychus truncatus Ehara, we screened specific primers of Tetranychus truncatus Ehara in RAPD amplification bands and transformed into stable SCAR markers. The results showed that there was a 717 bp specific band (GenBank accession number JF816665) amplified by primer OPB-04, and a pair of specific SCAR primers (Tt-303F and Tt -303R), PCR amplification conditions and procedures were optimized, a specific 303 bp DNA band was successfully amplified from different developmental stages of Tetranychus urticae and not amplified in other species The SCAR primer pair was also successfully verified in the field population of Tetranychus urticae collected from eggplant and kidney bean hosts in Beijing.