应用DNA条形码结合形态特征鉴定口岸船舶截获蛾类研究

Identification of moths intercepted by shipping quarantine using DNA barcoding and morphological characteristics

口岸船舶检疫截获昆虫之中,鳞翅目蛾类占较大比例。然而,截获的蛾类标本外部形态常磨损严重,加之涉及类群多,所需参考资料广泛、复杂,给口岸船舶检疫截获蛾类的鉴定工作带来了许多困难。本文以海南出入境检验检疫局八所口岸船舶检疫截获的18头蛾类作为研究材料,通过DNA条形码技术批量初筛与外生殖器特征鉴定核查相结合的方法,对其进行鉴定。其中,10头蛾类标本通过线粒体COⅠ基因片段序列比对直接鉴定到种级水平,隶属于4科8属8种,经形态学特征检视复核,鉴定准确率为100%;其余8头标本依据线粒体COⅠ基因片段序列比对先鉴定到属,再进行外生殖器特征检视,鉴定为4科7属7种。本次鉴定出的15种蛾类之中,12种为全国口岸首次截获,3种为海南口岸首次截获。研究结果表明,DNA条形码技术批量初筛与形态特征鉴定复核相结合的鉴定方法,既能够解决传统形态鉴定对标本保存要求高、鉴定难度大的问题,有效提高检出率;同时又减少了检测周期,提高工作效率,为检验检疫部门截获蛾类的快速、准确鉴定提供重要参考。

Moths account for a great proportion of insects intercepted at the entry ports of countries. Unfortunately,the external morphological characteristics（including markings on the wings） of intercepted moth specimens are usually badly worn down,coupled with the fact that moths are also species-rich and occur in nearly all regions of the world and in a wide va riety of habitats. These challenges put forth higher requirements for the identification of intercepted moths,so extensive,complex references and identification processes were utilized to identify each moth＇s taxonomic rank. Insect morphological characteristics and DNA barcoding（mitochondrial COⅠ gene fragment sequence information） were examined to identify 18 suspect moth specimens intercepted at Basuo port in Hainan province. Among them,10 specimens were directly identified by mitochondrial CO Ⅰ gene fragment sequence information down to their species;belonging to 4 families,8 genera,and 8 species total. The DNA barcoding identification accuracy rate was 100% after double-checking the genitalia characteristics. The remaining 8 specimens were identified down to their genera according to their mitochondrial COⅠgene fragment sequence information,and then to their species by genitalia characteristics：results showed that they belonged to 4 families,7 genera,and 7 species total. Among the 15 species,12 species were initially intercepted at Chinese ports,while the remaining 3 species were initially intercepted at Hainan port. These results indicated that mitochondrial COⅠ gene fragment sequence information from insect DNA barcoding could significantly reduce the time spent on moth species identification and increase work efficiency. The results provide an important reference strategy for the rapid and accurate identification of moths in insect inspection and quarantine departments.