枣实蝇特异引物PCR鉴定技术  

Species-specific PCR Primers for Identification of Carpomyia vesuviana

作　　者：程晓甜[1] 阿地力.沙塔尔 张伟[2] 温俊宝[3] 李新泉[1] 陈梦 

机构地区：[1]新疆农业大学林学与园艺学院新疆教育厅干旱区林业生态与产业技术重点实验室,乌鲁木齐830052 [2]新疆出入境检验检疫局技术中心,乌鲁木齐830083 [3]北京林业大学省部共建森林培育与保护教育部重点实验室,北京100083 [4]新疆维吾尔自治区林业有害生物防治检疫局,乌鲁木齐830000

出　　处：《林业科学》2013年第11期98-102,共5页Scientia Silvae Sinicae

基　　金：新疆维吾尔自治区科技计划项目"特色林果重大病虫害持续高效绿色防控技术研究"(201130102-3);新疆维吾尔自治区重点学科森林培育资助项目

摘　　要：在设计的枣实蝇15对引物中，利用种特异引物PCR（SS—PCR）和琼脂糖凝胶电泳技术筛选出1对枣实蝇的特异性引物，引物的特异性用桔小实蝇、瓜实蝇、南瓜实蝇、番石榴实蝇和桃果实蝇等5种实蝇来验证。SS—PCR方法的检测灵敏度用40，20，10，1，0．1，0．01，0．001ng·μL^-1等7个不同浓度系列的枣实蝇DNA模板来检测。结果表明，SS．PCR方法的检测限度达0．01ng·μL^-1以下，最适模板DNA浓度为1～20ng·μL^-1。所设计的引物对枣实蝇不同虫态均能进行特异性扩增，结果准确可靠。该技术体系可用于枣实蝇的鉴定识别与检测监测，对阻止其进一步扩散蔓延具有重要意义。In this study SS-PCR and agarose gel electrophoresis technique were used to select a pair of specific primers from the 15 pairs of primers of designed for Carpomyia vesuviana. Five fruit fly species, Bactrocera dorsalis, B. cucurbitae, B. tau, B. correcta and B. zonata, were used to determine the primer specificity. A series of genomic DNA dilution of C. vesuviana, 40, 20, 10, 1 , 0. 1 , 0.01 , and 0. 001 ng·μL^-1, were used to detect the sensitivity of SS-PCR. The results showed that the detection limit of SS-PCR was less than 0.01 ng·μL^-1, and the optimum template DNA concentration was between 1 ng and 20 ng·μL^-1. The designed primers could be used for amplifying the DNA of C. vesuviana at different stages. The developed SS-PCR method can be used for identification, inspection and monitoring of C. vesuviana, which would have an important significance in preventing the insect from spreading any further.

关 键 词：种特异引物PCR 枣实蝇 琼脂糖凝胶电泳 鉴定 

分 类 号：Q965[生物学—昆虫学]