Evolutionary and structural analysis of the cytochrome c oxidase subunit I (COI) gene from Haematobia irritans, Stomoxys calcitrans and Musca domestica (Diptera: Muscidae) mitochondrial DNA.

This work describes the molecular characterization of the cytochrome c oxidase subunit I (COI) gene of the mitochondrial DNA from three species of great medical and veterinary importance: the horn fly, Haematobia irritans, the stable fly, Stomoxys calcitrans and the house fly, Musca domestica (Diptera: Muscidae) (Linnaeus). The nucleotide sequence in all species was 1536 bp in size and coded for a 512 amino acid peptide. The nucleotide bias for an A+T-rich sequence is linked to three features: a high A+T content throughout the entire gene, a high A+T content in the third codon position, and a predominance of A+T-rich codons. An anomalous TCG (serine) start codon was identified. Comparative analysis among members of the Muscidae, Scatophagidae, Calliphoridae and Drosophilidae showed high levels of nucleotide sequence conservation. Analysis of the divergent amino acids and COI protein topologies among these three Muscidae species agreed with the evolutionary model suggested for the insect mitochondrial COI protein. The characterization of the structure and evolution of this gene could be informative for further evolutionary analysis of dipteran species.     

The Mitochondrial Genome of Elodia flavipalpis Aldrich (Diptera: Tachinidae) and the Evolutionary Timescale of Tachinid Flies

Tachinid flies are natural enemies of many lepidopteran and coleopteran pests of forests, crops, and fruit trees. In order to address the lack of genetic data in this economically important group, we sequenced the complete mitochondrial genome of the Palaearctic tachinid fly Elodia flavipalpis Aldrich, 1933. Usually found in Northern China and Japan, this species is one of the primary natural enemies of the leaf-roller moths (Tortricidae), which are major pests of various fruit trees. The 14,932-bp mitochondrial genome was typical of Diptera, with 13 protein-coding genes, 22 tRNA genes, and 2 rRNA genes. However, its control region is only 105 bp in length, which is the shortest found so far in flies. In order to estimate dipteran evolutionary relationships, we conducted a phylogenetic analysis of 58 mitochondrial genomes from 23 families. Maximum-likelihood and Bayesian methods supported the monophyly of both Tachinidae and superfamily Oestroidea. Within the subsection Calyptratae, Muscidae was inferred as the sister group to Oestroidea. Within Oestroidea, Calliphoridae and Sarcophagidae formed a sister clade to Oestridae and Tachinidae. Using a Bayesian relaxed clock calibrated with fossil data, we estimated that Tachinidae originated in the middle Eocene.     

Structure and evolution of the mitochondrial genome of Exorista sorbillans: the Tachinidae (Diptera: Calyptratae) perspective

The first complete mitochondrial genome (mitogenome) of Tachinidae Exorista sorbillans (Diptera) is sequenced by PCR-based approach. The circular mitogenome is 14,960?bp long and has the representative mitochondrial gene (mt gene) organization and order of Diptera. All protein-coding sequences are initiated with ATN codon; however, the only exception is Cox I gene, which has a 4-bp ATCG putative start codon. Ten of the thirteen protein-coding genes have a complete termination codon (TAA), but the rest are seated on the H strand with incomplete codons. The mitogenome of E. sorbillans is biased toward A+T content at 78.4?%, and the strand-specific bias is in reflection of the third codon positions of mt genes, and their T/C ratios as strand indictor are higher on the H strand more than those on the L strand pointing at any strain of seven Diptera flies. The length of the A+T-rich region of E. sorbillans is 106?bp, including a tandem triple copies of a13-bp fragment. Compared to Haematobia irritans, E. sorbillans holds distant relationship with Drosophila. Phylogenetic topologies based on the amino acid sequences, supporting that E. sorbillans (Tachinidae) is clustered with strains of Calliphoridae and Oestridae, and superfamily Oestroidea are polyphyletic groups with Muscidae in a clade.     

The mitochondrial genome of the quiet-calling katydids, <Emphasis Type="Italic">Xizicus fascipes</Emphasis> (Orthoptera: Tettigoniidae: Meconematinae)

To help determine whether the typical arthropod arrangement was a synapomorphy for the whole Tettigoniidae, we sequenced the mitochondrial genome (mitogenome) of the quiet-calling katydids, Xizicus fascipes (Orthoptera: Tettigoniidae: Meconematinae). The 16,166-bp nucleotide sequences of X. fascipes mitogenome contains the typical gene content, gene order, base composition, and codon usage found in arthropod mitogenomes. As a whole, the X. fascipes mitogenome contains a lower A+T content (70.2%) found in the complete orthopteran mitogenomes determined to date. All protein-coding genes started with a typical ATN codon. Ten of the 13 protein-coding genes have a complete termination codon, but the remaining three genes (COIII, ND5 and ND4) terminate with incomplete T. All tRNAs have the typical clover-leaf structure of mitogenome tRNA, except for tRNA(Ser(AGN)), in which lengthened anticodon stem (9 bp) with a bulged nuleotide in the middle, an unusual T-stem (6 bp in constrast to the normal 5 bp), a mini DHU arm (2 bp) and no connector nucleotides. In the A+T-rich region, two (TA)n conserved blocks that were previously described in Ensifera and two 150-bp tandem repeats plus a partial copy of the composed at 61 bp of the beginning were present. Phylogenetic analysis found: i) the monophyly of Conocephalinae was interrupted by Elimaea cheni from Phaneropterinae; and ii) Meconematinae was the most basal group among these five subfamilies.     

白纹佛蝗线粒体全基因组序列

通过长PCR扩增线粒体全基因组进行保守引物步移法结合克隆测序技术,对白纹佛蝗mtDNA 全序列进行了测定和分析.结果表明:白纹佛蝗线粒体基因组全长15 657 bp,包含13 个蛋白编码基因、22个tRNA 基因和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11 202 bp,1 486 bp,2 156 bp 和 728 bp.37个基因的位置与飞蝗的一致,有9对基因间存在41 bp重叠,重叠碱基数在 1～8 bp之间;基因间隔序列共计21处 126 bp,间隔长度从 1～20 bp不等,最大的基因间隔是20 bp,是在tRNALys 和 ATP8 基因之间.还对lrRNA和srRNA二级结构进行了预测,同时也对tRNA反密码子臂的碱基对类型以及不同链上蛋白编码基因的A/T,C/G组成偏向性进行了详细的讨论.     

残锷线蛱蝶线粒体基因组全序列及其系统学意义(英文)

对残锷线蛱蝶(Parathymasulpitia)(鳞翅目:蛱蝶科)线粒体基因组全序列进行了测定。结果表明:残锷线蛱蝶线粒体基因组全序列全长为15268bp,除了在trnS1(AGN)和trnE基因之间有一段121bp长的基因间隔外,其基因的排列顺序及排列方向与大多数已测鳞翅目物种基本一致。在蛋白质编码基因中,除cox1以CGA作为其起始密码子之外,其余12个蛋白质编码基因都以标准的ATN作为起始密码子。此外,除nad4基因以单独的T为终止密码子,其余12个蛋白质编码基因都以TAA结尾。除trnS1(AGN)缺少DHU臂之外,22个tRNA基因都显示典型的三叶草形二级结构。除A+T富集区外的非编码序列中,线粒体基因组共含有11个基因间隔区。其中,最长的一个121bp的基因间隔区位于trnS1(AGN)和trnE之间,其A+T含量高达100%。另外,和其他鳞翅目物种一样,在其A+T富集区的3’端有一段长达18bp的poly-T结构。A+T富集区内部没有明显的小卫星样多拷贝重复序列,而含有一些微卫星样的重复结构。本研究基于13种蛋白编码基因序列的组合数据,用最大似然法和贝叶斯法对蛱蝶科几个主要亚科间共9个代表物种间的系统发生关系进行了分析。结果表明,本研究的结果与前人的分子系统学研究结论基本吻合(其中,线蛱蝶亚科和釉蛱蝶亚科互为姐妹群),而与形态学的研究结论不一致。     

Mitochondrial DNA of the blowflyPhormia regina: restriction analysis and gene localization

A study of an invertebrate mitochondrial genome, that of the blowflyPhormia regina, has been initiated to compare its structural and functional relatedness to other metazoan mitochondrial genomes. A restriction map of mitochondrial DNA (mtDNA) isolated from sucrose gradient-purified mitochondria has been established using a combination of single and double restriction endonuclease digestions and hybridizations with isolated mtDNA fragments, revealing a genome size of 17.5 kilobases (kb). A number of mitochondrial genes including those encoding the 12 S and 16 S ribosomal RNA, the cytochromec oxidase I subunit (COI) and an unidentified open reading frame (URF2) have been located on thePhormia mtDNA by Southern blot analysis using as probes both isolated mtDNA fragments and oligonucleotides derived from the sequences of previously characterized genes from rat andDrosophila yakuba mtDNAs. These data indicate that for those regions examined, the mitochondrial genome organization of blowfly mtDNA is the same as that ofDrosophila yakuba, the order being COI-URF2-12 S-16 S. These data also report the presence of an A + T-rich region, located as a 2.5-kb region between the URF2 and the 12 S rRNA genes, and its amplification by the polymerase chain reaction is described.     

Complete mitochondrial genome of Chilo suppressalis (Walker) (Lepidoptera: Crambidae)

The complete sequence of the mitochondrial genome (mitogenome) of the rice stem borer Chilo suppressalis (Walker) (Lepidoptera: Crambidae) was determined to be 15,465 bp. It contains 13 protein-coding genes (PCGs), 22 tRNA genes, the large and small rRNA genes, and an A+T-rich region. The nucleotide composition of the mitogenome of C. suppressalis is highly A+T biased, accounting for 79.70% in whole mitogenome, 77.74% in PCGs, 84.70% in tRNAs, 81.20% in rRNAs and 94.19% in A+T-rich region, respectively. The PCGs have typical ATN start codons, except for cox1, which contains the unusual CGA. The C. suppressalis A+T-rich region contains a conserved structure combining the motif ATAGA and a 19-bp poly-T stretch, but absence of the 9-bp poly-A element upstream trnM.     

The mitochondrial genome of Ruspolia dubia (Orthoptera: Conocephalidae) contains a short A+T-rich region of 70 bp in length.

The complete sequence (14 971 bp) of the Ruspolia dubia mitochondrial genome was determined and annotated. The genome contains the gene content, base composition, and codon usage typical of metazoan mitochondrial genomes. All 37 genes are conserved in the positions observed most frequently in insect mitochondrial genome structures. The secondary structures of both small subunit and large subunit rRNA were predicted. The most unusual features found were the initiation codon (TTA) of COI and a short A+T-rich region of 70 bp in length. In addition, a short, highly conserved polythymidine stretch that was previously described in Orthoptera and Diptera was also present in the A+T-rich region.     

萧氏松茎象线粒体基因组全序列测定与分析

象甲是鞘翅目中物种最丰富的类群, 目前关于其线粒体基因组全序列的研究还未见报道。本研究利用长距PCR和引物步移法对萧氏松茎象Hylobitelus xiaoi Zhang线粒体基因组全序列进行了测定。结果显示： 萧氏松茎象线粒体基因组序列全长16 123 bp（GenBank登录号为JX847496）, 共编码37个基因和1个非编码的控制区, 基因次序与典型的六足动物线粒体基因排列一致, 未发现基因重排现象。在基因组中两个值得注意的发现分别是： 1）N链上存在1个额外的trnV-like序列, 反密码子为GAC, 长度为69 bp, 其中65 bp与J链上的trnD重叠; 2）trnS^UCN和nad1之间存在1个长度为232 bp的基因间隔区。全部13个蛋白质编码基因的起始密码子均为ATN, 9个蛋白质编码基因的终止密码子为TAA, 其余4个蛋白质编码基因中, nad1和cox2的终止密码子为TAG, nad4和nad5则以不完整的终止密码子T作为终止信号。除trnS^AGN外, 其余的tRNAs均可形成典型的三叶草结构。而trnS^AGN的反密码子由TCT替代GCT, 反密码子臂延长形成9 bp（中间含1个碱基突起）, TΨC臂由正常的5 bp变为6 bp, DHU臂缩短仅1 bp, 各个臂之间没有连接碱基。线粒体控制区中包括10处长度不少于5 bp的poly-T（最长poly-T长度为14 bp）和2处微卫星样重复序列 (TA)₆和(TA)₉。本研究结果为探讨象甲总科在鞘翅目中的系统学地位及其与其他总科间的系统发生关系等问题提供了重要的分子生物学数据。     

The complete mitochondrial genome of grouse locust Tetrix japonica (Insecta: Orthoptera: Tetrigoidea)

We have determined the complete mitochondrial genome of a species of grouse locust, Tetrix japonica. The total length of the T. japonica mitogenome is 15,128 bp with 75.57% A+T content. It consists of 13 protein-coding, 22 transfer RNA (tRNA), and 2 ribosomal RNA (rRNA) genes, and an A+T-rich region. The A+T-rich region was located between the small rRNA and tRNA-Ile genes and is 531 bp in length.     

The mitochondrial genome of the common cattle grub,Hypoderma lineatum

The mitochondrial DNA of the cattle grub Hypoderma lineatum (de Villers) (Diptera: Oestridae) was completely sequenced. The entire molecule was 16 354 bp long and presented a heavy bias towards A + T, which accounted for 77.8% of the whole genome. Hypoderma lineatum genes were organized in the same order and orientation as in the mitochondrial genomes available for other species belonging to the Oestroidea superfamily and compared in this study [Chrysomya putoria (Wiedemann), Cochliomyia hominivorax (Coquerel), Lucilia sericata (Meigen) and Dermatobia hominis (L.)], except for the occurrence of a 102‐bp non‐coding region partially present in other species. The complete sequence of H. lineatum will represent a useful dataset to evaluate the evolutionary pattern of mtDNA within Oestroidea by using molecular information in diagnostic, taxonomic and evolutionary studies.     

Mitochondrial genome nucleotide substitution pattern between domesticated silkmoth, Bombyx mori, and its wild ancestors, Chinese Bombyx mandarina and Japanese Bombyx mandarina

Bombyx mori and Bombyx mandarina are morphologically and physiologically similar. In this study, we compared the nucleotide variations in the complete mitochondrial (mt) genomes between the domesticated silkmoth, B. mori, and its wild ancestors, Chinese B. mandarina (ChBm) and Japanese B. mandarina (JaBm). The sequence divergence and transition mutation ratio between B. mori and ChBm are significantly smaller than those observed between B. mori and JaBm. The preference of transition by DNA strands between B. mori and ChBm is consistent with that between B. mori and JaBm, however, the regional variation in nucleotide substitution rate shows a different feature. These results suggest that the ChBm mt genome is not undergoing the same evolutionary process as JaBm, providing evidence for selection on mtDNA. Moreover, investigation of the nucleotide sequence divergence in the A+T-rich region of Bombyx mt genomes also provides evidence for the assumption that the A+T-rich region might not be the fastest evolving region of the mtDNA of insects.     

云斑车蝗线粒体基因组全序列测定与分析

采用长距 PCR 扩增及保守引物步移法并结合克隆测序测定并注释了云斑车蝗 Gastrimargus marmoratus （Thunberg）的线粒体基因组全序列。结果表明：云斑车蝗线粒体基因组全序列为15 904 bp（GenBank登录号为EU527334）,A+T含量略高于非洲飞蝗Locusta migratoria,为76.04%,包括13个蛋白质编码基因,22个tRNA 基因,2个rRNA基因和一段1 057 bp的A+T富集区。蛋白质基因的起始密码子中,除COⅠ和ND5为TTG以外,均为昆虫典型的起始密码子ATN。ND5基因使用了不完全终止密码子T,其余基因均为典型的TAA或TAG。预测了22个tRNA基因的二级结构,发现tRNA^Ser（AGN）缺少DHU臂, tRNA^Ser（UGY）的反密码子环上有9个碱基。预测了云斑车蝗12S和16S rRNA二级结构,分别包括3个结构域30个茎环和6个结构域44个茎环。A+T富集区含有3个串联重复序列。     

Complete mitochondrial genome of the groundhopper Alulatettix yunnanensis (Insecta: Orthoptera: Tetrigoidea)

In this work, we sequenced the first complete mitochondrial genome of Tetrigoidea species, Alulatettix yunnanensis. The circle genome (15,104 bp) consists of 13 protein-coding, 22 transfer RNA, and 2 ribosomal RNA genes, and an A+T-rich region. It has the typical invertebrate mitochondrial gene arrangement.     

Complete mitochodrial genome of the crab spider Ebrechtella tricuspidata (Araneae: Thomisidae): A novel tRNA rearrangement and phylogenetic implications for Araneae

Mitochondrial genomes are widely used for phylogenetic and phylogeographic analyses among arthropods, but there is a lack of sufficient mitochondrial genome sequence data for spiders. Herein, we sequenced and characterized the complete mitochondrial genome of a crab spider Ebrechtella tricuspidata (Araneae: Thomisidae). The circular mitochondrial genome is 14,352 bp long, including a standard set of 37 genes and an A + T-rich region. Nucleotide composition is highly biased toward A + T nucleotides (77.3%). A novel gene order rearrangement is detected by a tRNA (trnL1) translocation. Tandem repeats are not identified in the A + T-rich region. Most of the tRNAs are greatly reduced in size and cannot be folded into typical cloverleaf-shaped secondary structures. The phylogenetic analysis confirms that the mitochondrial genome sequences are useful in resolving higher-level relationship of Araneae. Overall, our data present in this study will elevate our knowledge on the architecture and evolution of spider mitochondrial genome.     

Variation of the mitochondrial genome in the evolution of Drosophila

The evidence on mitochondrial genome variation and its role in evolution of the genus Drosophila are reviewed. The mitochondrial genome is represented by a circular double-stranded DNA molecule 16 to 19 kb in length. The genome contains no introns involved in recombination. The entire mitochondrial genome can be arbitrarily divided into three parts: (1) protein-coding genes; (2) genes encoding rRNA and tRNA; and (3) the noncoding regulatory region (A + T region). The selective importance of mutations within different mtDNA regions is therefore unequal. In Drosophila, the content of the A + T pairs in mtDNA is extremely low and a pattern of nucleotide substitution is characterized by a low transition/transversion ratio (and a low threshold of mutation saturation). The deletions and duplications are of common occurrence in the mitochondrial genome. However, this genome lacks such characteristic for the nuclear genome aberrations as the inversions and transpositions. The phenomena of introgression and heteroplasmy provide an opportunity to study the adaptive role of the mitochondrial genome and its role in speciation. Analysis of evidence concerning mtDNA variation in different species of the genus Drosophila made it possible to ascertain data on phylogenetic relationships among species obtained by studying nuclear genome variation. In some species, mtDNA variation may serve as a reliable marker for population differentiation within a species, although evidence on the population dynamics of the mtDNA variation is very scarce.     

The phylogeny of Orthoptera inferred from mtDNA and description of Elimaea cheni(Tettigoniidae:Phaneropterinae)mitogenome

The complete 15,831 bp nucleotide sequence of the mitochondrial genome from Elimaea cheni(Phaneropterinae)was determined.The putative initiation codon for cox1 was TTA.The phylogeny of Orthoptera based on different mtDNA datasets were analyzed with maximum likelihood(ML)and Bayesian inference(BI).When all 37 genes(mtDNA)were analyzed simultaneously,the monophyly of Caelifera and Ensifera were recovered in the context of our taxon sampling.The phylogeny of Orthoptera was largely consistent with previous phyl...     

Electron microscope heteroduplex study of Drosophila mitochondrial DNAs: evolution of A+T-rich region.

Mitochondrial DNAs (mtDNA) from three species of the genus Drosophila (D. melanogaster, D. simulons, and D. virilis) were compared by electron microscope heteroduplex mapping. Analysis of heteroduplex molecules revealed that the A + T-rich region of these mtDNAs has undergone quite extensive base sequence divergence, whereas the remainder of the molecule was found to share apparently complete base sequence homology in all three species. The differences in the sizes of the A + T-rich regions, as determined from the heteroduplex measurements, completely account for the differences in the total sizes of these mtDNAs. A segment of approximately 0.1 kb is conserved within the A + T-rich regions of D. simulans and D. virilis mtDNAs, but not within the A + T-rich region of D. melanogaster mtDNA. HaeIII restriction endonuclease analysis of the heteroduplex molecules has further shown that the unique HaeIII site of D. virilis mtDNA molecule is apparently conserved in both D. melanogaster and D. simulans mtDNA molecules. Finally, electrophoretic patterns of HaeIII-digested mtDNAs from all three species were found to be different and distinguishable from each other suggesting that single base substitutions have probably taken place throughout the entire mitochondrial genome.