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本研究选择线粒体DNA(mtDNA)细胞色素氧化酶Ⅰ基因(COⅠ)为分子标记基因,以双翅目实蝇科昆虫DNA序列为目标,建立了我国进境植物检疫害虫地中海实蝇Ceratitis capitata、芒果小条实蝇C.cosyra和纳塔尔小条实蝇C.rosa等生物芯片检测方法。地中海实蝇及其近缘种检测芯片由检测探针(实蝇科通用探针1条,小条实蝇属通用探针1条,地中海实蝇、芒果小条实蝇和纳塔尔小条实蝇近缘种探针2条和种特异探针4条)、质控探针(定位点探针、阳性质控、阴性质控和空白对照探针各1条)组成。芯片检测结果表明,检测探针特异性强,能实现上述3种实蝇的种类快速区分和准确鉴定;检测方法稳定性好,地中海实蝇不同虫态(卵、幼虫、蛹和成虫)和不同地理种群检测结果完全一致。地中海实蝇生物芯片检测技术将为我国进口果蔬中检疫性实蝇快速筛查和种类鉴定提供检测方法,同时,还可应用到其他属的实蝇以及相关害虫的检疫中,为有害生物的快速鉴定提供了新方法。
In this study, mitochondrial DNA (mtDNA) cytochrome oxidase Ⅰ gene (COⅠ) was selected as the molecular marker gene. Based on the DNA sequences of the Diptera tetrafidae, we established the phytotoxin-producing pests such as Ceratitis capitata, C.cosyra, small fruit fly and Natal strip fly C.rosa bio-chip detection methods. Detection of Mediterranean fruit flies and its related species chips by the detection probe (Progenitor common probe 1, a small real fruit genus generic probe 1, the Mediterranean fruit fly, mango small fruit fly and Natal strip Fruit fly relatives probe 2 and species-specific probe 4), quality control probe (positioning point probe, positive quality control, negative quality control and blank control probe each one). The chip test results showed that the specificity of the detection probe was strong, and the species of the three fruit flies could be quickly distinguished and accurately identified. The detection method was stable, and the different morphologies (eggs, larvae, pupae and adult) of the Mediterranean fruit fly were different Geographical population test results are exactly the same. The detection technology of the fruit fly biochip will provide the detection method for the rapid screening and species identification of the imported fruit and vegetable quarantine fruit flies in China. At the same time, it can also be applied to the quarantine of fruit flies and related pests of other genera, Identification provides a new way.