基于DNA条形码对粘虫四种近缘种的鉴定与亲缘关系分析

Identification and phylogenetic analysis of four closely related species of the oriental armyworm Mythimna separata based on DNA barcoding

为开发快速、准确的粘虫Mythimna separata(Walker)近源种鉴定技术,通过线粒体COΙ基因DNA条形码技术对在野外采集的4种粘虫近缘种——白点粘夜蛾Leucania loreyi、淡脉粘夜蛾L.roseilinea、瘠粘夜蛾L.pallidior和虚研夜蛾Aletia pseudaletiana的COI基因片段进行序列分析,并对4种粘虫近缘种同粘虫和Gen Bank中其它6种粘虫近缘种之间的亲缘关系进行了分析。结果表明,克隆得到白点粘夜蛾、淡脉粘夜蛾、瘠粘夜蛾和虚研夜蛾大小为658 bp的COΙ基因片段,与粘虫COI基因片段的核苷酸序列同源性依次为93.31%、92.86%、91.95%和90.88%,遗传距离分别为0.067、0.071、0.081和0.091;所编码的219个氨基酸序列同源性依次为100.00%、100.00%、99.09%和99.09%。与其它6种粘虫近缘种的COI基因片段序列分析中,白点粘夜蛾和淡脉粘夜蛾与粘虫、一点粘虫Mythimna unipuncta、仿劳粘夜蛾L.insueta、白脉粘夜蛾L.venalba聚为1个亚群,亲缘关系较近;瘠粘夜蛾和虚研夜蛾与白杖研夜蛾A.lalbum、角线研夜蛾A.conigera、秘夜蛾M.turca聚为1个亚群,亲缘关系较近。表明4种近缘种与粘虫的亲缘关系由远到近依次为白点粘夜蛾、淡脉粘夜蛾、粘瘠粘夜蛾和虚研夜蛾。

In order to study the relationship of four field-collected closely related species of the oriental armyworm Mythimna separata and other six closely related species in the GenBank, CO1 barcode se- quences of the four armyworm species were analyzed. The results showed that the CO1 gene fragments of the four species of Noctuidae were all 658 bp in length. The CO1 sequences of Leucania loreyi, L. roseilinea, L. pallidior and Aletia pseudaletiana shared 93.31%, 92.86%, 91.95% and 90.88% identity with that of M. separata, respectively, and 100.00%, 100.00%, 99.09% and 99.09% in amino acid se- quences. The pairwise distances between the C01 gene sequences of the four Noctuidae species and M. separata were 0.067, 0.071, 0.081 and 0.091, respectively. In addition, the C01 gene fragments of the four closely related species and other six armyworm sibling species were analyzed, and the results showed that L. loreyi and L. roseilinea had a closer phylogenetic relationship with M. separata, M. uni- puncta, L. insueta, and L. venalba; L. pallidior andA. pseudaletiana had a closer relationship with A. lal-bum, A. conigera and M. turca. Taking the genetic distance analysis, homologous identity analysis and phylogenetic tree analysis together, the four species L. loreyi, L. roseilinea, L. pallidior and A. pseudale- tiana had closer genetic relationship with M. separata, in order.