摘要
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。
The present study eatablished genechip technique for Ceratitis capitata, C. rosa and C. cosyra, in which the mitochondrial cytochrome oxidase Ⅰ（CO Ⅰ ） gene was used as molecular marker, and the DNA sequences of fruit flies （Diptera： Tephritidae） were used as target sequence. The genechip detection system for fruit flies was composed of detecting probes and quality-control probes for C. capitata. The detecting probes included one family probe, one genus probe, three related species probes and four species specific-probes, and the quality-control probes included anchor probe, positive probe, negative probe and blank probe, with one for each. The results of gene chip showed that the specificity of detecting probes was specific to distinguish the three Ceratitis fruit flies. The detection method showed good repeatability and stability. The results of different stages （egg, pupa, larva, adult） and different geographical provenances of C. capitata were completely consistent. The genechip technique for Ceratitis fruit flies provided rapid detection and species identification method to quarantine fruit flies from imported fresh fruits and vegetables in China. Meanwhile, the technique also could be used in detecting other tephritid and related pests in plant quarantine.
出处
《昆虫学报》
CAS
CSCD
北大核心
2008年第1期61-67,共7页
Acta Entomologica Sinica
基金
科技部奥运科技(2008)行动计划资助项目(2004BA904B06-6)
关键词
实蝇科
近缘种
基因芯片
分类鉴定
Tephritidae
Ceratitis capitata
related species
genechip
taxonomic identification
