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首页> 外文期刊>DNA Barcodes >COI gene geographic variation of Gypsy moth (Lepidoptera: Lymantriidae) and a TaqMan PCR diagnostic assay
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COI gene geographic variation of Gypsy moth (Lepidoptera: Lymantriidae) and a TaqMan PCR diagnostic assay

机译：吉普赛蛾（鳞翅目：Lymantriidae）的COI基因地理变异和TaqMan PCR诊断分析

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Gypsy moth, an important forest/urban pest worldwide, is separated into the European and Asian subspecies, and has important quarantine significance. Diagnostic technique that can accurately and quickly distinguish subspecies is lacking. This study aimed to evaluate genetic difference between the subspecies, and subsequently to develop a reliable and high throughput molecular based diagnostic tool for distinguishing the subspecies. COI genes of 25 gypsy moth samples from China, Russia, Mongolia, Japan and the United States were sequenced. DNASTAR analysis revealed that gypsy moth COI gene was 1531bp long. The UPGM phylogenetic tree constructed based on the COI gene indicated that European subspecies (U.S. population) and Asian subspecies were distinctively divided into two branches. Japanese populations had a far distantly relationship with other Asian populations forming a separate branch. There was a single base substitution (base transformation only) at 14 consistent locations between Asian and American populations, but 13 of them coded the same amino acid. A MGB proper and TaqMan assay was designed base on the base substitution at 406th bp that coded a different amino acid. This allele typing assay took only 4 hours and could accurately distinguish gypsy moth subspecies of Europe and Asia. The study enriches the knowledge basis of genetic differentiation of gypsy moth subspecies. And more importantly the TaqMan assay is the first report of such diagnostic tool that could deliver rapid and accurate results and suitable for routine quarantine inspections to distinguish Asian and European gypsy moth subspecies. This study was supported by the Ministry of Science and Technology of the People’s Republic of China (Science and technology supporting project: 2012BAK11B03; International cooperation project: 2009DFA31950) and Jiangsu Entry and Exit Inspection and Quarantine Bureau (2014KJ45).
机译：吉普赛蛾是一种重要的全球性森林/城市害虫，被分为欧洲和亚洲的亚种，并具有重要的检疫意义。缺乏能够准确，快速区分亚种的诊断技术。这项研究旨在评估亚种之间的遗传差异，随后开发出一种可靠的，高通量的基于分子的诊断工具来区分亚种。对来自中国，俄罗斯，蒙古，日本和美国的25个吉普赛蛾样品的COI基因进行了测序。 DNASTAR分析显示，吉普赛蛾COI基因长1531bp。基于COI基因构建的UPGM系统树表明，欧洲亚种（美国种群）和亚洲亚种被明显地分为两个分支。日本人口与其他亚洲人口形成了一个独立的分支，有着非常遥远的关系。在亚裔和美洲人群之间的14个一致位置上有一个碱基取代（仅碱基转化），但其中13个编码相同的氨基酸。根据第406 bp的碱基取代（设计了不同的氨基酸），设计了MGB适当的TaqMan测定法。这种等位基因分型测定仅用了4个小时，就可以准确地区分欧洲和亚洲的吉普赛蛾亚种。该研究丰富了吉普赛蛾亚种遗传分化的知识基础。更重要的是，TaqMan测定法是这种诊断工具的第一份报告，该工具可提供快速而准确的结果，适合常规检疫检查以区分亚洲和欧洲的吉普赛蛾亚种。这项研究得到了中华人民共和国科学技术部的支持（科学技术支持项目：2012BAK11B03;国际合作项目：2009DFA31950）和江苏省出入境检验检疫局（2014KJ45）。

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