导　　师： 尹文英

学科专业： 071002

授予学位： 博士

作　　者： ;

机构地区： 中国科学院研究生院

摘　　要： 实蝇科Tephritidae 昆虫是双翅目Diptera 的大科之一,全世界已描述的种类超过450 属4300 余种,其中包括地中海实蝇Ceratitis capitata 在内的70 余种是农业上有重要经济意义的害虫。实蝇的鉴定是国际果蔬贸易中最突出的技术问题之一,建立检疫性实蝇特别是未成熟虫态快速可靠的鉴定方法在植物检疫上有重要的意义。 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mtDNA COI 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TaqMan MGB 探针4 条,用于检疫性实蝇的分子生物学快速鉴定研究。主要研究结果如下: I.首次建立了SYBR Green 实时荧光PCR 快速鉴定检疫性实蝇的方法。利用自行设计的特异引物,应用SYBR Green 实时荧光PCR 技术,分别建立了SYBR Green 实时荧光PCR/(SG-PCR/)快速鉴定地中海实蝇C. capitata、杨桃实蝇B. carambolae、番石榴实蝇B. correcta、桔小实蝇B. dorsalis、辣椒实蝇B. latifrons、芒果实蝇B. occipitalis、木瓜实蝇B. papayae、菲律宾实蝇B. philippinensis、瓜实蝇B. cucurbitae 和南瓜实蝇B. tau 等10 种检疫性实蝇的方法,SYBR Green 实时荧光PCR 产物的特异性通过溶解曲线分析/(MCA/)来确认。 II.在SYBR Green 实时荧光PCR 方法获得的特异引物的基础上,分别建立了种特异引物PCR/(SS-PCR/)快速鉴定10 种检疫性实蝇的方法。 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 鉴定 克隆

分 类 号： [S41]

领　　域： [农业科学]