大豆蚜细胞色素氧化酶Ⅱ基因的克隆及其在捕食性天敌昆虫鉴定中的应用

Cloning and sequencing of cytochrome oxidase Ⅱ gene of Aphis glycines and its application in detecting natural enemies

利用通用引物,对大豆蚜Aphis glycinesMatsumura的细胞色素氧化酶Ⅱ(COⅡ)基因序列进行克隆和测序。测得的序列长度为810 bp,并与已在GenBank上登录的其他蚜虫COⅡ基因序列进行比较分析,验证其同源性,同时将该序列在GenBank进行了登记(登录号为DQ265743)。根据此片段的碱基序列设计了1对大豆蚜特异引物,其扩增片段约为270 bp;种特异性检验结果表明,该引物只对大豆蚜具有扩增能力,对其他相关蚜虫种类不具有扩增效果;并用此引物对大豆蚜捕食性天敌进行定性检测,结果表明,在取食过大豆蚜的异色瓢虫Harmonia axyridis、龟纹瓢虫Propylaeajaponica、大草蛉Chrysopa septempunctata幼虫以及小花蝽Orius similes成虫和幼虫等捕食性昆虫的中肠中均能检测到大豆蚜的DNA片段;而在未取食蚜虫的上述天敌中却未能扩增出来。

Cytochrome oxidase subunit Ⅱ gene segment of Aphis glycines was cloned and sequenced. The length of the sequence was 810 bp other aphids registered at GenBank primer was developed, and its this pair of primer showed the （GenBank accession no. DQ265743） to identify their molecular homology. amplified single band was about 270 bp. ~ The sequence was contrasted with One pair of sequence-characterized The specificity tests performed with presence of the band only in A. glycines. Molecular identification of the natural enemies fed on A. glycines was tested with the specific primer. The results indicated that the target DNA fragment of A. glycines was detected in guts of the predatory insects Harmonia axyridis, Propylaea japonica, Chrysopa septempunctata, and Orius similes, but not detected in the above predatory insects without feeding A. glycines.