›› 2017, Vol. 60 ›› Issue (11): 1324-1338.doi: 10.16380/j.kcxb.2017.11.011

• 研究论文 • 上一篇    下一篇

小红珠绢蝶线粒体基因组特征及基于线粒体基因组的蝶类高级阶元系统发育关系分析

张敏1,*, 赵盼1,2, 尹洁1, 李涛1, 张婷婷1, 曹天文3, 马恩波   

  1. (1. 山西大学应用生物学研究所, 太原 030006; 2. 山西大学生命科学学院, 太原 030006; 3. 山西省农业科学院植物保护研究所, 太原 030031)
  • 出版日期:2017-11-20 发布日期:2017-11-20

Characterization of the complete mitochondrial genome of Parnassius nomion (Lepidoptera: Parnassiidae) and analysis of the higher-level phylogenetic relationships of butterflies based on mitochondrial genome

ZHANG Min1,*, ZHAO Pan1,2, YIN Jie1, LI Tao1, ZHANG Ting-Ting1, CAO Tian-Wen3, MA En-Bo1   

  1.  (1. Institute Applied Biology, Shanxi University, Taiyuan 030006, China; 2. College of Life Science, Shanxi University, Taiyuan 030006, China; 3. Institute of Plant Protection, Shanxi Academy of Agricultural Sciences, Taiyuan 030032, China)
  • Online:2017-11-20 Published:2017-11-20

摘要: 【目的】了解小红珠绢蝶Parnassius nomion线粒体基因组的特征,并从线粒体基因组水平探究蝶类高级阶元的系统发育关系。【方法】采用PCR扩增技术及Sequencher 4.8拼接软件获得小红珠绢蝶线粒体基因组全序列。参考鳞翅目昆虫已知线粒体基因组全序列并使用MEGA6.0软件对小红珠绢蝶线粒体基因组中各基因进行定位和注释。采用tRNA Scan-SE 1.21在线预测小红珠绢蝶线粒体基因组tRNA基因的二级结构。基于线粒体基因组13个蛋白质编码基因的核苷酸序列重建了包含凤蝶总科中凤蝶科(Papilionidae)、绢蝶科(Parnassiidae)、粉蝶科(Pieridae)、眼蝶科(Satyridae)、蛱蝶科(Nymphalidae)、灰蝶科(Lycaenidae)、斑蝶科(Danaidae)、珍蝶科(Acraeidae)、喙蝶科(Libyheidae)和蚬蝶科(Riodinidae)10个科28种蝴蝶的系统发育关系。【结果】结果表明,小红珠绢蝶线粒体基因组全序列总长度为15 362 bp (GenBank登录号: MF496134),包含13个蛋白质编码基因、22个tRNA基因、2个rRNA基因和1个A+T富含区。小红珠绢蝶线粒体基因组中存在较高的A+T含量(79.6%)。小红珠绢蝶线粒体基因组13个蛋白质编码基因中UUA的相对同义密码子使用频率(RSCU)最高(5.08),而AGG和CCG相对同义密码子使用频率(RSCU)均较低(0),这与大紫蛱蝶Sasakia charonda coreana的分析结果一致。在所测得的22个tRNA基因中,除tRNASer(AGN)缺少DHU臂外,其余tRNA基因均能形成典型的三叶草结构,这与鳞翅目中目前已得到的其他昆虫线粒体基因组中tRNA基因的二级结构一致。系统发育分析结果显示,凤蝶总科内蚬蝶科与灰蝶科的亲缘关系最近;粉蝶科与蛱蝶科、珍蝶科、眼蝶科、斑蝶科、喙蝶科、蚬蝶科和灰蝶科的系统发育关系更近;绢蝶科与凤蝶科锯凤蝶亚科亲缘关系最近,随后二者与凤蝶亚科物种聚为一支。在绢蝶科中,小红珠绢蝶与依帕绢蝶Parnassius epaphus的亲缘关系最近。【结论】本研究支持绢蝶科物种归为绢蝶亚科,绢蝶亚科、锯凤蝶亚科和凤蝶亚科归入凤蝶科,且绢蝶亚科与锯凤蝶亚科为姐妹群。

关键词:  鳞翅目, 凤蝶总科, 小红珠绢蝶, 线粒体基因组, 系统发育

Abstract: 【Aim】 The objective of this study is to understand the characteristics of the complete mitochondrial genome of Parnassius nomion and to explore the higherlevel phylogenetic relationships of butterflies based on mitochondrial genome sequences.【Methods】 The PCR amplification technique and Sequencher 4.8 software were used to obtain the whole sequence of mitochondrial genome of P. nomion. Based on the known complete mitochondrial genomes of lepidopteran species, MEGA6.0 software was used to locate and annotate each gene of P. nomion mitochondrial genome. tRNA Scan-SE 1.21 was used to predict the secondary structure of the tRNA genes online. Based on the nucleotide sequences of 13 protein-coding genes of mitochondrial genome, the phylogenetic relationships among 28 butterflies of 10 families (Papilionidae, Parnassiidae, Pieridae, Satyridae, Nymphalidae, Lycaenidae, Danaidae, Acraeidae, Libytheidae and Riodinidae) of Papilionoidea were reconstructed. 【Results】 The results indicated that the complete mitochondrial genome of P. nomion is a circular molecule of 15 362 bp (GenBank accession no.: MF496134), including 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes and an A+T-rich region. The mitochondrial genome of P. nomion is biased toward a high A+T content (79.6%). The relative synonymous codon usage (RSCU) of UUA is the highest in the 13 protein-coding genes in the mitochondrial genome of P. nomion, the relative synonymous codon usage value is 5.08, but AGG and CCG has a relatively low RSCU of 0, which is consistent with the analysis results of Sasakia charonda coreana. All 22 tRNA genes show the classic clover-leaf structure, except that the dihydrouridine (DHU) arm of tRNASer (AGN) forms a simple loop, which is consistent with the secondary structure of tRNA genes in the mitochondrial genomes of other lepidopteran insects. The results of phylogenetic analysis showed that Riodinidae and Lycaenidae are sister groups, Pieridae is closer with the clade of (Lycaenidae+Riodinidae)+(((Nymphalidae+Acraeidae)+Satyridae+Danaidae)+Libyheidae), Parnassiidae is a sister group of Zerynthiinae, and then Parnassiidae+Zerynthiinae is clustered with Papilioninae. P. nomion is closely related with Parnassius epaphus in Parnassiidae. 【Conclusion】 Our study supports that Parnassiidae species should be included in Parnasiinae, Parnasiinae, Zerynthiinae and Papilioninae all belong to Papilionidae, and Parnasiinae and Zerynthiinae are sister groups.

Key words: Lepidoptera, Papilionoidea, Parnassius nomion, mitochondrial genome, phylogeny