Characterization of the complete mitochondrial genome of Hydrotaea spinigera (Diptera: Muscidae) with phylogenetic implications

Hydrotaea spinigera Stein is a muscid species with high forensic importance. To further supplement the genome-level features of related species, the complete mitochondrial genome of H. spinigera is amplified, sequenced, annotated, analyzed, and compared with 13 other species of the family Muscidae. This mitogenome is 15,517?bp in length, with a standard set of 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA) genes, two ribosomal RNA (rRNA) genes, and a typical non-coding control region. The gene organization, base composition, and secondary structures of tRNA genes of this mitogenome are similar to other dipteran species. Phylogenetic analyses based on all the mitochondrial PCGs and rRNA genes for 14 muscid and one outgroup taxa are performed using Bayesian inference (BI) and maximum likelihood (ML) analysis. The inferred trees indicate that the Muscidae, the subfamily Muscinae, and the tribes Azeliini and Reinwardtiini are monophyletic, whereas the monophyly of the subfamily Azeliinae and the tribe Muscini is not supported.     

Phylogenetic analyses of the harvest mouse,Micromys minutus (Rodentia: Muridae) based on the complete mitogenome sequences

Harvest Mouse (Micromys minutus) has a very wide range of distribution in Asia and Europe. However, the phylogenetic relationship of M. minutus is still uncertain. In this study, we determined the complete mitochondrial (mt) genome sequences of M. minutus, and used the complete mitochondrial genome sequences constructed the phylogenetic tree of Muroidea. The size of the genome is 16,232 bp in length and has a base composition of 33.6% A, 29.1% T, 24.8% C, and 12.5% G. The mitogenome structure was similar to that of typical vertebrate and other rodents' mitochondrial genomes, includes 13 protein-coding genes, 2 rRNA genes (12S rRNA and 16S rRNA), 22 tRNA genes, and 1 control region. We suggested a new initiation codon for ND5 (NADH dehydrogenase subunit), which has been never reported in the mitochondrial genome of vertebrate. The ML and BI phylogenetic trees, which based on the combination of the 12 protein-coding genes, supported strongly that the genus Micromys was represent an early offshoot within the Muridae with high support values (BI = 1.00, ML = 100).     

Complete mitochondrial genome of the Eurasian flying squirrel Pteromys volans (Sciuromorpha, Sciuridae) and revision of rodent phylogeny

In this study, the complete mitochondrial genome of the Eurasian flying squirrel Pteromys volans (Rodentia, Sciuromorpha, Sciuridae) was sequenced and characterized in detail. The entire mitochondrial genome of P. volans consisted of 16,513 bp and contained 13 protein-coding genes, 22 tRNA genes, two rRNA genes, and two non-coding regions. Its gene arrangement pattern was consistent with the mammalian ground pattern. The overall base composition and AT contents were similar to those of other rodent mitochondrial genomes. The light-strand origin generally identified between tRNA ^Asn and tRNA ^Cys consisted of a secondary structure with an 11-bp stem and an 11-bp loop. The large control region was constructed of three characteristic domains, ETAS, CD, and CSB without any repeat sequences. Each domain contained ETAS1, subsequences A, B, and C, and CSB1, respectively. In order to examine phylogenetic contentious issues of the monophyly of rodents and phylogenetic relationships among five rodent suborders, here, phylogenetic analyses based on nucleotide sequence data of the 35 rodent and 3 lagomorph mitochondrial genomes were performed using the Bayesian inference and maximum likelihood method. The result strongly supported the rodent monophyly with high node confidence values (BP 100 % in ML and BPP 1.00 in BI) and also monophylies of four rodent suborders (BP 85–100 % in ML and BPP 1.00 in BI), except for Anomalumorpha in which only one species was examined here. Also, phylogenetic relationships among the five rodent suborders were suggested and discussed in detail.     

The Complete Mitochondrial Genomes of Three Bristletails (Insecta: Archaeognatha): The Paraphyly of Machilidae and Insights into Archaeognathan Phylogeny

The order Archaeognatha was an ancient group of Hexapoda and was considered as the most primitive of living insects. Two extant families (Meinertellidae and Machilidae) consisted of approximately 500 species. This study determined 3 complete mitochondrial genomes and 2 nearly complete mitochondrial genome sequences of the bristletail. The size of the 5 mitochondrial genome sequences of bristletail were relatively modest, containing 13 protein-coding genes (PCGs), 2 ribosomal RNA (rRNA) genes, 22 transfer RNA (tRNA) genes and one control region. The gene orders were identical to that of Drosophila yakuba and most bristletail species suggesting a conserved genome evolution within the Archaeognatha. In order to estimate archaeognathan evolutionary relationships, phylogenetic analyses were conducted using concatenated nucleotide sequences of 13 protein-coding genes, with four different computational algorithms (NJ, MP, ML and BI). Based on the results, the monophyly of the family Machilidae was challenged by both datasets (W12 and G12 datasets). The relationships among archaeognathan subfamilies seemed to be tangled and the subfamily Machilinae was also believed to be a paraphyletic group in our study.     

The complete mitochondrial genome of Eterusia aedea (Lepidoptera,Zygaenidae) and comparison with other zygaenid moths

Zygaenidae comprises >1036 species, including many folivorous pests in agriculture. In the present study, the complete mitochondrial genome (mitogenome) of a major pest of tea trees, Eterusia aedea was determined. The 15,196-bp circular genome contained the common set of 37 mitochondrial genes (including 13 protein-coding genes, two rRNA genes, and 22 tRNA genes) and exhibited the similar genomic features to reported Zygaenidae mitogenome. Comparative analyses of Zygaenidae mitogenomes showed a typical evolutionary trend of lepidopteran mitogenomes. In addition, we also investigated the gene order of lepidopteran mitogenomes and proposed that the novel gene order trnA-trnR-trnN-trnE-trnS-trnF from Zygaenidae and Gelechiidae and most other gene rearrangements of this tRNA cluster evolved independently. Finally, the mitogenomic phylogeny of Lepidoptera was reconstructed based on multiple mitochondrial datasets. And all the phylogenetic results revealed the sister relationships of Cossoidea and Zygaenoidea with both BI and ML methods, which is the first stable mitogenomic evidence for this clade.     

两种毛蚜线粒体基因组比较分析

【目的】线粒体基因组分析已被应用于昆虫系统发育研究。本研究以蚜科Aphididae重要类群毛蚜亚科物种为代表,测定并比较分析了该类蚜虫的线粒体基因组特征,探讨了基于线粒体基因组信息的蚜虫系统发育关系重建。【方法】以毛蚜亚科三角枫多态毛蚜Periphyllus acerihabitans Zhang和针茅小毛蚜Chaetosiphella stipae Hille Ris Lambers,1947为研究对象,利用长短PCR相结合的方法测定线粒体基因组的序列,分析了基因组的基本特征;基于在线t RNAscan-SE Search Server搜索方法预测了t RNA的二级结构;基于12个物种(本研究获得的2个物种和10个Gen Bank上下载的物种数据)的蛋白编码基因(PCGs)序列,利用最大似然法和贝叶斯法重建了蚜科的系统发育关系。【结果】两种毛蚜均获得了约94%的线粒体基因组数据,P.acerihabitans获得了14 908 bp,控制区为1 205 bp;C.stipae获得了13 893 bp,控制区为609 bp。两种毛蚜同时获得33个基因,包含接近完整的13个蛋白编码基因(PCGs)(nad5不完整),18个tRNA,2个rRNA基因;ka/ks值表明,C.stipae的进化速率更快。从基因组组成、基因排列顺序、核苷酸组成分析、密码子使用情况、t RNA二级结构等特征来分析,两种蚜虫线粒体基因组基本特征相似。系统发育重建结果表明毛蚜亚科、蚜亚科的单系性得到了支持,毛蚜亚科位于蚜科的基部位置。【结论】两种毛蚜线粒体基因组的基本特征相似,符合蚜虫线粒体基因组的一般特征,两种线粒体基因组的长度差异主要来自控制区长度的不同;系统发育重建支持毛蚜亚科与蚜亚科的单系性,毛蚜亚科位于蚜科较为基部的位置。研究结果为蚜虫类系统发育重建提供了参考。     

The complete mitochondrial genome of Parasesarma pictum (Brachyura: Grapsoidea: Sesarmidae) and comparison with other Brachyuran crabs

Mitochondrial DNA (mtDNA) is an extrachromosomal genome which can provide important information for evolution and phylogenetic analysis. In this study, we assembled a complete mitogenome of a crab Parasesarma pictum (Brachyura: Grapsoidea: Sesarmidae) from next generation sequencing reads at the first time. P. pictum is a mudflat crab, belonging to the Sesarmidae family (subfamily Sesarminae), which is perched on East Asia. The 15,716 bp mitogenome covers 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), 2 ribosomal RNA genes (rRNAs), and one control region (CR). The control region spanns 420 bp. The genome composition was highly A+T biased 75.60% and showed negative AT-skew (−0.03) and negative GC-skew (−0.47). Compared with the ancestor of Brachyura, the gene order of Sesarmidae has several differences and the gene order of P. pictum is typical for mitogenomes of Sesarmidae. Phylogenetic tree based on nucleotide sequences of mitochondrial 13 PCGs using BI and ML determined that P. pictum has a sister group relationship with Parasesarma tripectinis and belongs to Sesarmidae.     

Mitochondrial genome of the cockscomb pearl mussel Cristaria plicata (Bivalvia, Unionoida, Unionidae)

The complete mitochondrial genome sequence of the cockscomb pearl mussel Cristaria plicata, which is an endangered species in South Korea, was sequenced. The circle genome (15,708 bp in size) consists of 13 protein-coding genes, 2 rRNA genes, and 22 tRNA genes. There were 26 noncoding regions (NCs) found throughout the mitogenome of C. plicata, ranging in size from 2 to 327 bp, and the two largest NC regions, NC286 and NC326, were found between ND5 and tRNA(Gln) (286 bp) and between tRNA(Glu) and ND2 (326 bp), respectively. The 13 mitochondrial protein-coding genes of a female individual of C. plicata collected from Korea (15,708 bp) were compared to those of the Chinese individual (15,712 bp) published before. The result showed that ND3 is the most conserved with 100% nucleotide similarity, and each of the other protein-coding genes has ca. 99%, respectively. The two largest NCs among 26 NCs have totally 98% nucleotide similarity between Korean and Chinese ones.     

Complete mitochondrial genome of a Chinese scorpion Mesobuthus martensii (Chelicerata, Scorpiones, Buthidae).

The complete mitochondrial genome (15,034 bp) of a Chinese scorpion Mesobuthus martensii (Buthidae) was sequenced and characterized in detail. The genome contains 13 protein-coding genes, 21 transfer RNA genes, two ribosomal RNA genes and a large non-coding region ( = CR). Its gene arrangement pattern is identical to that of Limulus polyphemus (Chelicerata, Xiphosura), with the exceptions of the tRNA(Glu)-tRNA(Ile)-tRNA(Met) (Q-I-M) arrangement and tRNA(Asp)-loss. Additional interesting features are found and discussed: high frequency of Leu(UUG) codon use, low A+T content of the genome (66.75%), and six repeat units (five 60-nt-long and one 58-nt-long repeats) in the 998-nt CR. Bayesian analysis based on amino acid sequences of the 12 proteincoding genes (excluding ATP8) reveals that the family Buthidae (Order Scorpiones) and the class Arachnida form strong monophyletic groups within Chelicerata, respectively. It indicated that the scorpions are the most ancestral arachnids.     

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 phylogenetie hypotheses.Rhaphidophoridae to be a sister group of Tettigoniidae,and the relationships among four subfamilies of Tettigoniidae were(Phaneropterinae+(Conocephalinae+(Bradyporinae+Tettigoniinae))).Pyrgomorphidae was the most basal group of Caelifera.The relationships among six acridid subfamilies were(Oedipodinae+(Acridinae+(Gomphocerinae+(Oxyinae+(Calliptaminae +Cyrtacanthaeridinae))))).However,we did not recover a monophyletic Grylloidea.Myrmecophilidae clustered into one clade with Gryllotalpidae instead of with Gryllidae.ML and BI analyses of all protein coding genes(using all nucleotide sequence data or excluding the third codon position,and amino acid sequences)revealed a topology identical to that of the entire mtDNA genome dataset.However,22 tRNAs genes excluding the DHU loop and T()C loop(TRNA),and two rRNA genes(RRNA)perform poorly when analyzed as single dataset.Our results suggest that the best phylogenetie inferences were ML and BI methods based on total mtDNA.Excluding tRNA genes,rRNA genes and the third codon position of protein coding genes from dataset and converting nucleotide sequences to amino acid sequences do not positively affect phylogenetic reconstruction.     

土蜂科线粒体基因组序列测定和分析

【目的】本研究旨在通过针尾部(Aculeata)昆虫线粒体基因组系统发育分析认知土蜂科(Scoliidae)的单系性及系统发育位置。【方法】利用Illumina Hiseq2500二代测序技术测序土蜂科3属5种的线粒体基因组,并进行注释和分析;基于针尾部昆虫36个线粒体基因组13个蛋白质编码基因(protein-coding genes, PCGs)和2个rRNA基因序列采用最大似然法(maximum likelihood, ML)和贝叶斯法(Bayesian inference, BI)法构建系统发育树。【结果】新测序的土蜂科5个线粒体基因组为五带波壁土蜂Colpa quinquecincta线粒体基因组(GenBank登录号：OM103696),齿石波壁土蜂Colpa tartara线粒体基因组(GenBank登录号：OM103697),厚大长腹土蜂Megacampsomeris grossa线粒体基因组(GenBank登录号：OM103796),台湾大长腹土蜂Megacampsomeris formosensis线粒体基因组(GenBank登录号：OM142776)和斯式土蜂Sc...     

Complete genomic sequence of the filamentous nitrogen-fixing cyanobacterium Anabaena sp. strain PCC 7120.

The nucleotide sequence of the entire genome of a filamentous cyanobacterium, Anabaena sp. strain PCC 7120, was determined. The genome of Anabaena consisted of a single chromosome (6,413,771 bp) and six plasmids, designated pCC7120alpha (408,101 bp), pCC7120beta (186,614 bp), pCC7120gamma (101,965 bp), pCC7120delta (55,414 bp), pCC7120epsilon (40,340 bp), and pCC7120zeta (5,584 bp). The chromosome bears 5368 potential protein-encoding genes, four sets of rRNA genes, 48 tRNA genes representing 42 tRNA species, and 4 genes for small structural RNAs. The predicted products of 45% of the potential protein-encoding genes showed sequence similarity to known and predicted proteins of known function, and 27% to translated products of hypothetical genes. The remaining 28% lacked significant similarity to genes for known and predicted proteins in the public DNA databases. More than 60 genes involved in various processes of heterocyst formation and nitrogen fixation were assigned to the chromosome based on their similarity to the reported genes. One hundred and ninety-five genes coding for components of two-component signal transduction systems, nearly 2.5 times as many as those in Synechocystis sp. PCC 6803, were identified on the chromosome. Only 37% of the Anabaena genes showed significant sequence similarity to those of Synechocystis, indicating a high degree of divergence of the gene information between the two cyanobacterial strains.     

绿眼赛茧蜂线粒体基因组全序列测定和分析

【目的】测定绿眼赛茧蜂Zele chlorophthalmus线粒体基因组全序列,分析其基因组结构及茧蜂科(Braconidae)部分类群的系统发育关系。【方法】利用Illumina MiSeq二代测序技术对绿眼赛茧蜂的线粒体基因组进行测序,对基因组序列进行拼装、注释,分析其结构特点和碱基组成;基于22种茧蜂科昆虫的COX1蛋白编码基因序列,应用最大似然法(ML)和邻接法(NJ)构建系统发育树,分析绿眼赛茧蜂与茧蜂科其他昆虫的系统发育关系。【结果】绿眼赛茧蜂线粒体基因组全长16 661 bp(GenBank登录号: MG822749),包含13个蛋白质编码基因、22个tRNA基因和2个rRNA基因,共37个基因,以及1个控制区。线粒体基因组有明显的核苷酸组成的偏倚,AT偏正,GC偏负,其A+T含量为82.83%。基因排列顺序与推测的昆虫祖先的序列不完全一致,tRNA基因7处发生重排。13个蛋白质编码基因均以ATN为起始密码子,以TAA为终止密码子。在22个tRNA基因二级结构中,除tRNA^His(H)缺失TΨC环和tRNA^Cys(C)仅剩二氢尿嘧啶(DHU)臂和反密码子臂外,其余tRNA基因均能形成典型的三叶草结构。基于COX1蛋白编码序列的系统发育分析结果显示,与绿眼赛茧蜂亲缘关系最近的是同属于赛茧蜂属的雪跗赛茧蜂Z. niveitarsis。【结论】本研究首次获得绿眼赛茧蜂线粒体基因组全序列。结果表明绿眼赛茧蜂隶属于优茧蜂亚科(Euphorinae)赛茧蜂属,并支持赛茧蜂属的单系性。     

The complete mitochondrial genome of the Asiatic cavity-nesting honeybee Apis cerana (Hymenoptera: Apidae)

In the present study, we determined the complete mitochondrial DNA (mtDNA) sequence of Apis cerana, the Asiatic cavity-nesting honeybee. We present here an analysis of features of its gene content and genome organization in comparison with Apis mellifera to assess the variation within the genus Apis and among main groups of Hymenoptera. The size of the entire mt genome of A. cerana is 15,895 bp, containing 2 ribosomal RNA genes, 13 protein-coding genes, 22 transfer RNA (tRNA) genes and one control region. These genes are transcribed from both strands and have a nucleotide composition high in A and T. The contents of A+T of the complete genomes are 83.96% for A. cerana. The AT bias had a significant effect on both the codon usage pattern and amino acid composition of proteins. There are a total of 3672 codons in all 13 protein-coding genes, excluding termination codons. The most frequently used amino acid is Leu (15.52%), followed by Ile (12.85%), Phe (10.10%), Ser (9.15%) and Met (8.96%). Intergenic regions in the mt genome of A. cerana are 705 bp in total. The order and orientation of the gene arrangement pattern is identical to that of A. mellifera, except for the position of the tRNA-Ser(AGN) gene. Phylogenetic analyses using concatenated amino acid sequences of 13 protein-coding genes, with three different computational algorithms (NJ, MP and ML), all revealed two distinct groups with high statistical support, indicating that A. cerana and A. mellifera are two separate species, consistent with results of previous morphological and molecular studies. The complete mtDNA sequence of A. cerana provides additional genetic markers for studying population genetics, systematics and phylogeographics of honeybees.     

The mitochondrial genome of the predatory mite Metaseiulus occidentalis (Arthropoda: Chelicerata: Acari: Phytoseiidae) is unexpectedly large and contains several novel features

The complete mitochondrial genome of the phytoseiid Metaseiulus occidentalis (Arthropoda: Chelicerata: Acari: Phytoseiidae) has been sequenced. It is 24,961 bp in length and contains a 14,695-bp unique region, a 345-bp triplicated region, and a 9921-bp duplicated region, in that order. The A+T content of the unique region is 76.9% and contains 11 protein coding (COI-III; ATP6-8; CytB; ND1, 2, 4, 5 and 4L), two ribosomal RNA (srRNA and lrRNA), 22 transfer RNA (tRNA) genes, and two copies of D-loop control sequence. Two genes (ND3 and 6) appear to be missing, but there is a large intergenic spacer (390 bp) present, which could contain ND3 if a different codon usage is employed. The gene order is completely different from the pattern in all other known chelicerates, including the horseshoe crab Limulus polyphemus [Lavrov et al., Mol. Biol. Evol., 2000; 17:813-824]. All the inferred tRNA genes are missing the TPsiC arm, but this arm has fused with the variable arm to generate a TV replacement loop. The duplicated region (9921 bp) contains 18 genes in the same order as in the unique region from CytB to tRNA-His, plus one copy of D-loop control sequence (311 bp) and a partial tRNA-Leu2 sequence (34 bp). The small triplicated region (345 bp) contains a D-loop control sequence (311 bp) and a partial tRNA-Leu2 sequence (34 bp). Because of these anomalies, amplifying sequences posed technical difficulties, but were accomplished by using a primer-walking strategy and increasing the AT content to 75% in the high-fidelity PCR dNTP mix. This is the first phytoseiid mitochondrial genome to be completely sequenced and the largest (25 kb) detected from the Chelicerata.     

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.     

Complete mitochondrial genome of Pycnonotus xanthorrhous (Passeriformes,Pycnonotidae) and phylogenetic consideration

The complete mitochondrial genome (mitogenome) of Pycnonotus xanthorrhous was sequenced via next generation sequencing. The full length of the circular genome is 16,952 bp. It consists of 37 typical animal mitochondrial genes including 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA) and 2 ribosomal RNA (rRNA) genes. P. xanthorrhous also contains one control region (CR) and one pseudo-control region, and shares the identical gene arrangements with sequenced Pycnonotus spp. which differs from the typical vertebrates gene order. Phylogenetic analyses indicates that Passerida sensu stricto contains three major clades and the core Sylvioidea form a monophyletic group. Furthermore, we investigated the evolution of control region within this lineage and revealed the multiple independent origins of duplicate control region.