南部松齿小蠹和长林小蠹幼虫分子鉴定

Molecular identification for larvae of Ips grandicollis(Eichhoff) and Hylurgus ligniperda Fabricius

小蠹幼虫在口岸中经常被截获。应用DNA条形码技术,对厦门口岸分别从美国进口的南方黄松和从澳大利亚进口的辐射松截获的小蠹幼虫进行分子鉴定,选用引物CJ-J-2183/TL2-N-3014对线粒体COⅠ部分序列,引物S3660/A335对核糖体28S部分序列进行PCR扩增和测序,并与Gen Bank中对应的序列进行比对。结果表明,从美国南方黄松截获小蠹幼虫样品COⅠ部分序列和28S部分序列与NCBI数据库中南部松齿小蠹同源性达99%,从澳大利亚辐射松截获小蠹幼虫样品COⅠ部分序列和28S部分序列与NCBI数据库中长林小蠹同源性分别达100%和99%;基于COⅠ和28S部分序列构建的系统发育树,两种小蠹幼虫样品分别与南部松齿小蠹和长林小蠹最为接近,根据序列分析和系统发育关系分析,将截获的小蠹幼虫鉴定为南部松齿小蠹和长林小蠹。

The larvae of bark beetle are often intercepted at port. DNA barcodes can realize rapid identification of them. Partial sequences of COⅠ gene and 28 S r DNA gene using bark beetle larvae samples were amplified and sequenced with primers CJ-J-2183/TL2-N-3014 and S3660/A335. By blasting with known data in Gen Bank the result showed that the similarities of CO Ⅰ and 28 S between bark beetle sample from southern yellow pine of USA（Sample No：USA2） and I. grandicollis were 99%,the similarities of COⅠ and 28 S between bark beetle sample from monterey pine of Australian（Sample No：NZ4）and H. ligniperda were 100% and 99%.The molecular phylogenetic relationship of bark beetle based on CO Ⅰ gene sequences and 28 S gene sequences showed that USA2 and AF113351.1 I. grandicollis are the same clade,NZ4 and HM002621.1 H. ligniperda are the same clade. The larvae were identified as I.grandicollis and H. ligniperda.