麻竖毛天牛线粒体基因组测定及 天牛科线粒体基因组比较分析
Determination of the complete mitochondrial genome of Thyestilla gebleri and comparative analysis of the mitochondrial genome in Cerambycidae
杨 锦1** 曹利军2 耿涌鑫1 魏丹峰1 陈 敏1***
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DOI:10.7679/j.issn.2095-1353.2017.092
作者单位:1. 北京林业大学省部共建森林培育与保护教育部重点实验室,北京 100083; 2. 北京市农林科学院植物保护环境保护研究所,北京 100097
中文关键词: 麻竖毛天牛,天牛科,线粒体基因组,系统发育
英文关键词:Thyestilla gebleri, Cerambycidae, mitochondrial genome, phylogeny
中文摘要:
【目的】 线粒体基因组分析是研究昆虫系统发育的重要手段。本研究通过测定麻竖毛天牛Thyestilla gebleri(Faldermann,1835)线粒体基因组,比较分析天牛科Cerambycidae线粒体基因组的特征,进而初步探讨麻竖毛天牛系统发育地位和天牛科部分类群之间的系统进化关系。【方法】 采用引物步移法测定麻竖毛天牛线粒体基因组全序列。参照GenBank收录的16种天牛线粒体基因组序列进行基因注释;采用在线软件tRNAscan-SE Search Server对转运RNA(tRNA)的二级结构进行了预测。通过对16种天牛的线粒体基因组进行重新注释,结合本研究获得的麻竖毛天牛线粒体基因组进行序列特征比较分析。基于11个蛋白编码基因的核苷酸序列,利用最大似然法构建了天牛科17种天牛的系统发育树。【结果】 麻竖毛天牛线粒体基因组全长15 505 bp,A+T含量为74.07%,包含13个蛋白编码基因(PCGs),2个核糖体RNA(rRNA)基因,22个tRNA基因和一个长度为872 bp的控制区,未发现基因重排。通过比对17种天牛的线粒体基因组,发现长翅暗天牛Vesperus conicicollis一个tRNA(trn P)基因的移位。17种天牛的tRNA中,trnS1(AGN)的D臂均缺失,其余tRNA都具有典型的三叶草结构。大部分种类的蛋白编码基因起始密码子为典型的ATN(ATA、ATT、ATC、ATG),只有部分种类的Nad1、COI、ATP8基因存在特殊的起始密码子(TTG、AAC、AAT、GTG),终止密码子均为常见的TAR(TAA、TAG)或不完全的T和TA。系统发育树中,6个亚科分别单独分支,其中,麻竖毛天牛与云斑白条天牛Batocera lineolata聚为一支。【结论】 麻竖毛天牛线粒体基因组符合天牛线粒体基因组的一般特征;除少数tRNA基因存在重排外,天牛科线粒体基因排列相对稳定;基于线粒体基因组的系统发育分析支持天牛科6个亚科的单系性,麻竖毛天牛和云斑白条天牛亲缘关系较近。
英文摘要:
[Objectives] Analysis of mitochondrial genome is an important way to reveal the phylogenic relationship in insects. The aim of this study was to determinate the mitochondrial genome of Thyestilla gebleri and comparatively analyze the characteristics of mitochondrial genome in Cerambycidae. We also investigated the phylogenetic position of T. gebleri and the phylogenetic relationship among species in Cerambycidae based on mitochondrial genomes. [Methods] The mitochondrial
genome of T. gebleri was determined using primer walking method. The annotation of mitochondrial genes of T. gebleri were based on the GenBank data of mitochondrial genome in Cerambycidae. The secondary structure of transport RNA (tRNA) was predicted by online tRNAscan-SE Search Server. After re-annotating the mitochondrial genome of 16 species available in GenBank, we conducted comparative analysis of the mitochondrial genomic features for Cerambycidae. A phylogenetic tree for 17 species in Cerambycidae was constructed by Maximum Likelihood method based on the nucleotide sequences of 11 protein-coding genes. [Results] The total length of mitochondrial genome of T. gebleri was 15 505 bp with an A+T content of 74.07%. It contained 13 protein coding genes (PCGs), 2 ribosomal RNA (rRNA) genes, 22 tRNA genes and A 872 bp-long control region, and no gene rearrangement was found. The results revealed the translocation of trn P (one of a tRNA gene) in Vesperus conicicollis. The secondary structure of tRNA genes had the typical clover structure except the absent of “D” arm in trnS1(AGN). The start codon of protein-coding genes were typical ATN (ATA, ATT, ATC, ATG) except that several special start codon (TTG, AAC, AAT, GTG) existed in Nad1, COI, ATP8 in some species. The termination codon of all the protein coding genes was common TAR (TAA, TAG), T or TA. In the phylogenetic analysis, 6 subfamilies were clustered into separate branches, and T. gebleri was clustered together with Batocera lineolata. [Conclusion] T. gebleri has similar mitochondrial genome to those previously reported longicorn beetles. Except that several tRNA genes are rearranged, the mitochondrial genes in Cerambycidae are relatively stable. Phylogenetic reconstruction supported the monophyly of the six subfamilies and presented that T. gebleri was genetically close to B. lineolata in this study.