重大潜在入侵害虫番茄潜叶蛾的SS-COⅠ快速检测技术  

Species-specific CO Ⅰ primers for rapid identification of Tuta absoluta (Meyrick), a significant, potential alien species

作　　者：张桂芬[1,2] 刘万学[1,2] 郭建洋[1,2] 张毅波[1] 万方浩[1,2] 

机构地区：[1]中国农业科学院植物保护研究所,植物病虫害生物学国家重点实验室,北京100193 [2]农业部外来入侵生物预防与控制研究中心,北京100081

出　　处：《生物安全学报》2013年第2期80-85,共6页Journal of biosafety

基　　金：国家"973"计划项目(2009CB119200);公益性行业(农业)科研专项(201103026)

摘　　要：【背景】番茄潜叶蛾是源自南美洲的一种最具毁灭性的世界性入侵害虫,其适生区范围广,且与其他潜叶类昆虫难以准确、快速识别。【方法】以田间常见的其他6种潜叶类害虫为对照,采用基于mtDNA COⅠ基因的种特异性SS-COⅠ方法,研究番茄潜叶蛾快速分子检测技术。【结果】种特异性检测结果显示,SS-COⅠ引物只对番茄潜叶蛾mtDNA具有扩增能力,对美洲斑潜蝇、南美斑潜蝇、三叶草斑潜蝇、葱斑潜蝇、豌豆潜叶蝇、桃潜叶蛾等其他潜叶类害虫不具有扩增效果。该引物不仅对成虫和单粒卵具有很好的扩增效能,对单根触角、头部、胸部、腹部以及翅和足等成虫残体亦具有同样的扩增能力,其最低检出阈值为312.5pg·μL-1。【结论与意义】该方法在有效防范番茄潜叶蛾侵入我国,保障农业生产安全和生态环境安全中意义重大。[ Background]The tomato leafminer Tuta absoluta （Meyrick）, native to South America, is one of the most devastating and invasive pest species worldwide. Morphological identification of T. absoluta is limited by small size, the high degree of similarity and polymorphism. [ Method] In this study, a method was described for the development of DNA marker for identification of T. abso- luta. One pair of species-specific PCR primers （SS-COI） based on a fragment of known mitochondrial DNA cytochrome oxidase I （mtDNA COI） sequence was designed for T. absoluta. The specificity of the primer pair was validated using six other leafminer species, including Liriomyza sativae Blanchard, L. huidobrensis （Blanchard）, L. trifolli （Brugess）, L. chinensis （Kato）, Phytomyza horticola Goureau and Lyonetia clerkella L. , all of them commonly occurring in China. [ Result ] All T. absoluta specimens were detected, and no cross reactions with other leafminer species were observed. The method was tested on single egg and adult debris （including antenna, head, thorax, abdomen, wing, leg） of T. absoluta, and proved to be applicable. Even at the concentration of 312.5 pg · μL^-1 DNA, the target fragment could be detected in all replicates. [ Conclusion and significance ] The technique developed here should be useful in quarantine at ports of entry and in guarantee the safety of agricultural production and ecological environment in China.

关 键 词：番茄潜叶蛾 潜在入侵生物 种特异性SS-COⅠ引物 分子检测 快速鉴定 

分 类 号：S436.412.2[农业科学—农业昆虫与害虫防治] S41-30[农业科学—植物保护]