导　　师： 尹文英

学科专业： G1002

授予学位： 博士

作　　者： ;

机构地区： 中国科学院上海生命科学研究院植物生理生态研究所

摘　　要： 　　本项研究以国内外普遍关注的检疫性实蝇为研究对象，测定了果实蝇属bactrocera和小条实蝇属ceratitis共10亚属28种检疫性实蝇的mtdnacoi基因部分序列，获得32条基因片段(长约670bp)；对这些基因序列与已公开报道的其它同源序列进行比较分析，自行设计和筛选特异引物14对(亚属特异引物2对，种特异引物12对)和taqmanmgb探针4条，用于检疫性实蝇的分子生物学快速鉴定研究。 本研究建立的分子生物学方法不仅能鉴定实蝇的成虫而且能快速鉴定幼虫、蛹和卵等虫态，使实蝇的鉴定周期从传统的形态学鉴定所需20天时间缩短到2天以内，在植物检疫检疫性实蝇的日常鉴定中有极大的优势。 本研究搭建了出入境植物检疫性害虫分子生物学鉴定的技术平台，为逐步建立昆虫的分子鉴定体系奠定了基础。 Tephritidae fruit fly is one of the largest families of Diptera. There are about 4300 species in more than 450 genera throught the world. About 70 species of this family, including the Miditerranean fruit fly, Ceratitis capitata, are of the quarantine importance. Identification of fruit flies is one of critical questions in international commercial fruits and vegetables trade. It is imperative to establish rapid and reliable methods to identify quarantine fruit flies especially in immature stages in plant quarantine. In this study, 28 species were selected from10 subgenera of Bactrocera and Ceratitis genera, which are of quarantine importance. Partial mtDNA COI gene sequences /(~670bp/) of these fruit flies were sequenced. These fragments were analyzed and aligned with the homologues of others closely related sepecies reported previously. Two of subgenus-specific, 11 of species-specific primers and four TaqMan MGB probes were designed for identification of these quarantine fruit flies. The main results were: I . The real-time PCR technique using SYBR Green I dye was firstly introduced in rapid identification of quarantine fruit flies. In SYBR Green real-time PCR /(SG-PCR/) assay, ten fruit flies, Ceratitis capitata, B. carambolae, B. correcta, B. dorsalis, B. latifrons, B. occipitalis, B. papayae, B. philippinensis, B. cucurbitae and B. tau, were accurately

关 键 词： 检疫性实蝇 基因 实时荧光 探针实时荧光 分子生物学 快速鉴定

领　　域： [生物学] [农业科学] [农业科学] [农业科学]