花斑皮蠹和黑斑皮蠹(鞘翅目:皮蠹科)的快速分子鉴定

Rapid Molecular Identification of Trogoderma glabrum and Trogoderma variabile(Coleoptera:Dermestidae)

斑皮蠹属昆虫个体小,形态高度相似,仅依据外部形态难以鉴定。用线粒体细胞色素氧化酶亚基I(COI)基因的通用引物,分别以黑斑皮蠹(Trogoderma glabrum)、花斑皮蠹(Trogoderma variabile)的DNA为模板,进行PCR扩增、测序。比较二者COI的碱基序列,找出所有序列多态性位点及相应的限制性内切酶,最终选取限制性内切酶AflⅡ对二者进行酶切。结果表明,黑斑皮蠹的COI序列可被限制酶AflⅡ切开,形成535bp和347bp两部分,而花斑皮蠹COI片段中因不具有AflⅡ酶切位点而不能被切开,从而将二者成功区分,实现对黑斑皮蠹和花斑皮蠹的快速分子鉴定。

In this paper, DNA isolated from species of T. glabrum and T. variabile was amplified by polymerase chain reaction （PCR） ,the products were directly sequenced, analyzed, and digested with restriction enzyme Afl II. The results proved that COI sequences of T. glabrum could be digested into two parts： 535bp and 347 bp, but T. variabile was not. Thus, T. gla- bruin and T. variabile were identified accurately and efficiently.