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应用种特异性PCR技术快速鉴定窝梗天牛

发表时间:2015-11-15  浏览量:1945  下载量:669
全部作者: 郑斯竹,徐梅,杨晓军,安榆林
作者单位: 苏州出入境检验检疫局外来有害生物防控技术中心;江苏出入境检验检疫局
摘 要: 针对窝梗天牛(Arhopalus foveicollis)与其他幽天牛族昆虫难以准确快速识别的问题,以窝梗天牛为靶标,并以14种幽天牛为参照,采用基于线粒体DNA(mtDNA)细胞色素C氧化酶亚基Ⅰ(COI)基因序列的种特异性(species-specific COI,SS-COI)PCR方法,研究其快速分子检测技术。通过已知鞘翅目COI基因序列通用型引物J1718/N2191,获得窝梗天牛及其他14种天牛的COI基因序列,根据测序结果设计SS-COI引物1对(WGF1/WGR1),其扩增片段大小为254 bp. 种特异性检验结果指出,该引物只对窝梗天牛的COI基因可扩增出254 bp大小的片段。研究结果表明,该技术体系完全可用于窝梗天牛的准确识别及检测监测,对有效阻截其进入我国具有重要意义。
关 键 词: 森林保护学;窝梗天牛;幽天牛族;种特异性PCR;分子检测;快速鉴定
Title: Rapid identification of Arhopalus foveicollis (Cerambycidae: Asemini) by using species-specific PCR technique
Author: ZHENG Sizhu, XU Mei, YANG Xiaojun, AN Yulin
Organization: Technology Center for Prevention and Control of Alien Pests, Suzhou Entry-Exit Inspection and Quarantine Bureau; Jiangsu Entry-Exit Inspection and Quarantine Bureau
Abstract: In the present study, a method with Arhopalus foveicollis as target and 14 other kinds of species of Asemini as reference was described on the development of DNA markers for rapidly identifying A. foveicollis for the problem of fast recognition between species of Asemini and A. foveicollis. A pair of universal primers was designed based on mitochondrial DNA (mtDNA) cytochrome C oxidase subunit I (COI) gene sequences using species-specific COI (SS-COI). Using the universal primers (J1718/N2191) of Coleoptera, the mtDNA COI genes of A. foveicollis and its similar species were amplified and sequenced. And then one pair of SS-COI primers (WGF1/WGR1) was designed, which was amplified a single band of 254 bp. The results showed that only COI gene of A. foveicollis specimens were detected with 254 bp. The diagnostic PCR assay developed here provides a quick, simple and reliable molecular technique for the identification and monitoring of A. foveicollis, which will be useful in intercepting and blocking the further spreading of A. foveicollis.
Key words: forest protection; Arhopalus foveicollis; Asemini; species-specific PCR; molecular detection; rapid identification
发表期数: 2015年11月第21期
引用格式: 郑斯竹,徐梅,杨晓军,等. 应用种特异性PCR技术快速鉴定窝梗天牛[J]. 中国科技论文在线精品论文,2015,8(21):2279-2283.
 
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