菜粉蝶线粒体基因组的全序列测定和分析

Sequencing and analysis of the complete mitochondrial genome of Pieris rapae Linnaeus(Lepidoptera:Pieridae)

目前关于蝶类线粒体基因组全序列及其分子进化的研究还不多见。本研究通过长PCR和引物步移法对菜粉蝶Pieris rapae Linnaeus线粒体基因组全序列进行了测定和初步分析。结果表明:菜粉蝶线粒体基因组全长15157bp,包含13个蛋白编码基因、22个tRNA和2个rRNA基因以及1个非编码的控制区域,它们的长度分别是11196bp,1474bp,2093bp和393bp。37个基因的位置与已报道的其他蝶类基本一致,共有10对基因间存在总共59bp的重叠,重叠碱基数在1～35bp之间;基因间隔序列共计13处120bp,间隔长度1～46bp不等,最大的基因间隔46bp,位于tRNAIle和tRNAGln基因之间。另外,基于13个蛋白质编码基因的氨基酸序列,重建了基于蛋白质编码基因序列数据的11种代表性蝶类的NJ和MP系统树。结果表明:凤蝶类(包括凤蝶和绢蝶)为一大支系,粉蝶类、灰蝶类与蛱蝶类(包括蛱蝶、珍蝶)构成另一大支系。结果不支持粉蝶科与凤蝶科(包括凤蝶类和绢蝶类)构成单系群,却显示粉蝶科、灰蝶科和蛱蝶科的组合为单系群。

Up to now,the higher level taxonomic and phylogenetic studies about the butterflies are still limited and remain many controversial issues.In this study,the complete mitochondrial genome of Pieris rapae Linnaeus was sequenced by using long-PCR and conserved primer walking techniques,aiming at supporting more mitogenomic data to further clarify these problems.The results of genome characterization analysis showed that the mitogenome is 15 157 bp in size,containing 13 protein-coding genes,22 transfer RNA genes,2 ribosomal RNA,and an A ＋ T-rich region,with the length of 11 196 bp,1 474 bp,2 093 bp and 393 bp,respectively;all the 37 genes are arranged in the same orientations as those of other butterflies determined;there are 10 overlapping sequences totalling 59 bp（1-35 bp for each sequence） and 13 intergenic spacer sequences totalling 120 bp（1-46 bp for each sequence）,scattered throughout the genome,respectively,and the largest（46 bp） spacer region is located between the tRNAIleand tRNAGlngenes.The neighbor-joining（NJ） and the maximum parsimony（MP） phylogenetic trees of the eleven representative butterfly species were reconstructed based on the complete 13 protein coding gene sequence data,and the results showed that Papilionidae（including papilionids and parnassiids） is a monophyletic group,and Pieridae is closely related with Lycaenidae and Nymphalidae（including nymphalids and acraeids in this study）.