西花蓟马的SCAR 分子检测技术.docVIP

西花蓟马的SCAR分子检测技术 孟祥钦1，闵 亮1, 3，万方浩1, 2，周忠实1, 2，王文凯3，张桂芬1, 2, ( (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) 摘要：西花蓟马Frankliniella occidentalis (Pergande)是一种世界性入侵害虫，寄主范围广，危害严重，2003年首次在我国发生危害，并有进一步扩散蔓延的趋势。针对蓟马类害虫虫体微小、形态相似，难以准确快速区分的问题，采用特征序列扩增区域（SCAR）标记技术，以西花蓟马及与之同域发生的其他种类蓟马为对象，筛选出1对西花蓟马特异性引物（FOMF/FOMR），其扩增片段大小为320 bp。种特异性检测结果显示，该对引物只对西花蓟马的基因组DNA具有扩增能力，对同域发生的花蓟马F. intonsa (Trybom)、禾花蓟马F. tenuicornis (Uzel)、烟蓟马Thrips tabaci L.等41种蓟马不具有扩增效果。该对引物不仅对不同虫态的西花蓟马具有扩增能力，而且在西花蓟马发生地的寄主植物组织内亦检测到了其卵的存在。同时，该检测技术灵敏度高，对成虫的最低检出阈值为1/160头。本检测技术在口岸检疫以及花卉、蔬菜和种苗调运中的害虫检测和监测中具有重要意义。 关键词：西花蓟马；RAPD；SCAR；；中图分类号: Q966 文献标识码: A 文章编号: 0454-6296(2010)0-0000-00 SCAR marker for rapid identification of the western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) MENG Xiang-Qin1, MIN Liang1,3, WAN Fang-Hao1,2, ZHOU Zhong-Shi1,2, WANG Wen-Kai3, ZHANG Gui-Fen1,2,* (1. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China; 2. Center for Management of Invasive Alien Species, Ministry of Agriculture, Beijing 100081, China; 3. Agriculture College, Yangtze University, Jingzhou, Hubei 434025, China) Abstract: The western flower thirps, Frankliniella occidentalis (Pergande), is a worldwide pest, causing serious damage to a wide range of host plants. It was first reported in China in 2003, with a trend of further spreading. Most of thrips species are small in size and similar in morphology, which makes it hard to be distinguished quickly. In the present research, a pair of SCAR (sequence characterized amplified regions) primers (FOMF/FOMR) which are specific to western flower thrips was developed by using other nine familiar thrips species as the control. The fragment amplified by the primers FOMF/FOMR was 320 bp in length. Specificity tests performed with this pair of primers showed that all F. occidentalis specimens were detected positiv