The complete mitochondrial genome of Tuta absoluta (Lepidoptera: Gelechiidae) and genetic variation in two newly invaded populations in China

Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is a devastating invasive pest worldwide, causing severe damage to tomatoes. Recently, it has been recorded in the northwestern and southwestern parts of China. Here, the mitogenomes and genetic variation of two newly invaded T. absoluta populations in Xinjiang and Yunnan, were determined. The results showed that the complete mitogenome size of T. absoluta is 15298 bp for the individual from Xinjiang and 15296 bp for the individual from Yunnan, which were both longer than the reported mitogenome from Spain (15290 bp). The mitogenome sequences of individuals collected from three locations showed high levels of sequence similarity, except for 8 polymorphic sites, which were in genes cox2 (1 site), cox3 (2 sites), cob (1 site), atp6 (1 site), nad1 (2 sites) and nad5 (1 site). Tuta absoluta mitogenomes share many features with other 6 Gelechiidae mitogenomes, except for several differences in the start and stop codons of protein-coding genes and the length of intergenic spacers. Seven partial mitochondrial genes (cox1, cox2, cox3, atp6, cob, nad1, and nad5) were used for genetic variation analysis, and significant population differentiation was found between the two populations based on cox2, atp6, nad1, and nad5. The complete mitogenomes and sensitive mitochondrial gene markers reported here provide useful data for further population genetics study of this pest.     

The rearranged mitochondrial genome of Podagrion sp. (Hymenoptera: Torymidae), a parasitoid wasp of mantis

The complete sequence of the mitochondrial genome of Podagrion sp. (Hymenoptera: Torymidae) is described. The mitogenome was 15,845 bp in size, and contained typical sets of mitochondrial genes. The base composition of the Podagrion sp. mitogenome was also biased toward A + T bases (81.8%). The mitochondrial genome of Podagrion sp. has a weak AT skew (0.07) and a strong GC skew (?0.26). Podagrion sp. exhibits a novel rearrangement compared with the ancestral order, including six protein-coding genes (nad3, cox3, atp6, atp8, cox2 and cox1), which have inverted to the minor strand from the major strand. The A + T-rich region of Podagrion sp., which is located between trnN and trnI, have five tandem repeats. The apomorphic rearrangements, including the conserved block “cox3-atp6-atp8-cox2-cox1-nad5-nad4-nad4l-nad6-cob” and the special locations of trnV and trnA, were mapped onto the phylogeny of Proctotrupomorpha.     

Complete mitochondrial genome of Bactrocera ritsemai (Insecta: Tephritidae) and phylogenetic relationship with its congeners and related tephritid taxa

Bactrocera ritsemai is a dacine fruit fly found in Indonesia. We report here the complete mitogenome of this fruit fly from Lombok, Indonesia determined by Illumina MiSeq sequencing and its phylogenetic relationship with its congeners and related tephritid taxa. The whole mitogenome of B. ritsemai had a total length of 15,927 bp, comprising 37 genes – 13 protein-coding genes (PCGs), 2 ribosomal ribonucleic acid (rRNA) and 22 transfer ribonucleic acid (tRNA) genes – and a control region (D-loop). Of the PCGs, 6 (atp6, cob, cox2, cox3, nad4, nad4l) had ATG start codon, 4 (nad2, nad3, nad5, nad6) had ATT, and one each had ATA (nad1), GTG (atp8) and TCG (cox1). Seven PCGs (atp6, atp8, cox2, cox3, nad2, nad4l, nad6) had TAA stop codon, 3 (cob, nad3, nad4) had TAG, and 3 had incomplete stop codon (cox1 – TA; nad1, nad5 – T). The TΨC-loop of tRNA was absent in trnF while trnS1 lacked the DHU-loop. Phylogenetic analysis based on 15 mt-genes (13 PCGs + 2 rRNA genes) indicated B. ritsemai forming a sister group with B. umbrosa and the subgenus Bactrocera was monophyletic. The genera Bactrocera and Zeugodacus were monophyletic while the subfamilies Dacinae and Tephritinae were paraphyletic. A broader taxa sampling of the Tephritidae is needed to better elucidate the phylogenetics and systematics of the tribes and subfamilies of tephritid fruit flies.     

The complete mitochondrial genome of Mahanta tanyae compared with other zygaenoid moths (Lepidoptera: Zygaenoidea)

The complete mitochondrial genome (mitogenome) of Mahanta tanyae was sequenced and extensively compared with all seven additionally reported zygaenoid mitogenomes. The M. tanyae mitogenome is circular, double-stranded, and 15,323 bp long. Gene content, gene order, and orientation are all typical of Lepidoptera, despite the existence of gene rearrangements for some other zygaenoid mitogenomes. Comparative analyses further showed that the incomplete termination codon T is consistently recognized in the mitochondrial cox1, cox2 and nad4 genes of all zygaenoid species, as well as in the nad5 gene in two limacodid species. Among 13 protein-coding genes, nad6 exhibits the highest evolutionary rate. The structure for each tRNA is highly conserved, including loss of the dihydorouidine (DHU) arm in trnS1 (AGN), but remarkable nucleotide variation exists, primarily in the pseudouridine (TψC) loops. Interestingly, in four species of Zygaenidae, the anticodons for trnS1 (AGN) are consistently UCU, instead of the routinely used codon GCU, in all three species of Limacodidae. In the intergenic region between trnS2 and nad1, a short sequence before the motif “ATACTAA” is present in the M. tanyae mitogenome that is unique among reported zygaenoid mitogenomes. In the A + T-rich region between the motif “ATTTA” and the microsatellite (AT)_n element, some nucleotides were present for most zygaenoid mitogenomes, which is, to our knowledge, rare even in reported lepidopteran mitogenomes. Phylogenetic analyses based on the combined 37 mitochondrial genes confirmed the position of M. tanyae in Limacodidae of the Zygaenoidea.     

Complete mitogenome of Nycteribia allotopa Speiser, 1901 (Diptera,Hippoboscoidea, Nycteribiidae) and comparative analysis of mitochondrial genomes of Nycteribiidae

In recent years, the global pandemic of bat-associated pathogens has led to increasing attention on bat ectoparasites. Numerous studies have identified human-associated pathogens in Nycteribiidae, indicating their potential as vectors. In this study, the first complete sequencing of the mitochondrial genome of Nycteribia allotopa Speiser, 1901 was sequenced and analyzed. We also compared the mitochondrial sequences of N. allotopa with those available in the database for other Nycteribiidae species. The complete mitochondrial genome of N. allotopa was found to be 15,161 bp in size with an A + T content of 82.49%. Nucleotide polymorphism analysis of 13 protein-coding genes from five species of Nycteribiidae showed that nad6 exhibited the most significant variation, while cox1 was the most conserved. Furthermore, selection pressure analysis revealed cox1 to exhibit the strongest purifying selection, while atp8, nad2, nad4L, and nad5 showed slightly looser purifying selection. Pairwise genetic distances indicated that cox1 and cox2 were evolving comparatively slowly, whereas atp8, nad2, and nad6 were evolving comparatively quickly. Phylogenetic trees constructed using Bayesian inference and maximum likelihood methods demonstrated that all four families within the superfamily Hippoboscoidea clustered into one branch each, indicating their monophyly. N. allotopa was found to be most closely related to the same genus N. parvula. This study significantly enriches the molecular database for Nycteribiidae and provides invaluable reference data for future species identification, phylogenetic analysis, and exploration of their potential as vectors for human-associated pathogens.     

Characterization of the complete mitochondrial genome of Tryporyza incertulas, in comparison with seven other Pyraloidea moths

The complete 15,223-bp mitochondrial genome (mitogenome) of Tryporyza incertulas (Walker) (Lepidoptera: Pyraloidea: Crambidae) was determined, characterized and compared with seven other species of superfamily Pyraloidea. The order of 37 genes was typical of insect mitochondrial DNA sequences described to date. Compared with other moths of Pyraloidea, the A + T biased (77.0%) of T. incertulas was the lowest. Eleven protein-coding genes (PCGs) utilized the standard ATN, but cox1 used CGA and nad4 used AAT as the initiation codons. Ten protein-coding genes had the common stop codon TAA, except nad3 having TAG as the stop codon, and cox2, nad4 using T, TA as the incomplete stop codons, respectively. All of the tRNA genes had typical cloverleaf secondary structures except trnS1(AGN), in which the dihydrouridine (DHU) arm did not form a stable stem-loop structure. There was a spacer between trnQ and nad2, which was common in Lepidoptera moths. A 6-bp motif ‘ATACTA’ between trnS2(UCN) and nad1, a 7-bp motif “AGC(T)CTTA” between trnW and trnC and a 6-bp motif “ATGATA” of overlapping region between atp8 and atp6 were found in Pyraloidea moths. The A + T-rich region contained an ‘ATAGT(A)’-like motif followed by a poly-T stretch. In addition, two potential stem-loop structures, a duplicated 19-bp repeat element, and two microsatellites ‘(TA)₁₂’ and ‘(TA)₉’ were observed in the A + T-rich region of T. incertulas mitogenome. Finally, the phylogenetic relationships of Pyraloidea species were constructed based on amino acid sequences of 13 PCGs of mitogenomes using Bayesian inference (BI) and maximum likelihood (ML) methods. These molecular-based phylogenies supported the morphological classification on relationships within Pyraloidea species.     

Characterization of the complete mitochondrial genome of Japanagallia spinosa and Durgades nigropicta (Hemiptera: Cicadellidae: Megophthalminae)

In this study, we determined and analyzed the complete mitochondrial genomes (mitogenomes) of Japanagallia spinosa and Durgades nigropicta (Hemiptera: Megophthalminae). The circular genome were 15,655 bp long in J. spinosa (GenBank: KY123686) and 15,974 bp long in D. nigropicta (GenBank: KY123687). The J. spinosa and D. nigropicta mitogenomes both contained 37 genes and the gene order was similar to that in other leafhoppers. All of the protein-coding genes started with ATN. In the J. spinosa mitogenome, the nad3, nad4L, and cytb genes used TAG as a stop codon, the atp8 and nad1 genes used TGA, and the cox2 gene used a single T. However, in the D. nigropicta mitogenome, three genes used a single T as the stop codon, whereas the nad3 gene used TAG. We predicted the secondary structures of the rRNAs in J. spinosa and D. nigropicta. The secondary structure of rrnL comprised six domains (domain III is absent in arthropods) with 42 helices and that of rrnS comprised three structural domains with 26 helices. Comparisons of J. spinosa and D. nigropicta detected some differences in H577 and H673. We determined the structural organization of the control regions in the mitogenomes of leafhoppers, where three types of repeat regions were found in most. The phylogenetic relationships between J. spinosa and D. nigropicta with related lineages were reconstructed using Bayesian inference and maximum likelihood analyses. The monophyly of each superfamily considered in this study was confirmed by the clades in the phylogenetic tree. And in this study, Cicadellidae was resolved as monophyletic by the phylogenetic analysis. This mitogenome information for J. spinosa and D. nigropicta could facilitate future studies of mitogenomic diversity and the evolution of related insect lineages.     

Complete mitochondrial genome of the hemp borer,Grapholita delineana (Lepidoptera: Tortricidae): Gene variability and phylogeny among Grapholita

The mitochondrial genome (mitogenome) has been extensively used in phylogenetics and species-level evolutionary investigations. The lepidopteran family Tortricidae (leaf-roller moths), including the genus Grapholita, contains numerous species of economic importance, but for the majority of Grapholita species, their mitogenomes remain poorly studied. Here, we sequence and annotate the full mitogenome of Grapholita delineana, an important pest of hemp worldwide and compare it with the mitogenomes of two congeneric species available from GenBank. The G. delineana mitogenome is 15,599 bp long, including 37 typical mitochondrial genes and an A + T-rich region. Gene content, order and orientation are identical to other reported tortricid mitogenomes. Analyses of nucleotide diversity, Ka/Ks, genetic distance and number of variable sites together suggest that nad6 is the fastest-evolving gene among the mitochondrial PCGs of Grapholita. Our analyses indicate that Grapholita, as presently defined, is not monophyletic, confirming previous morphological and multiple-gene studies, using mitogenomic evidence. Our study provides information on comparative mitogenomics of Tortricidae especially Grapholita.     

The complete mitogenome of Bombyx mori strain Dazao (Lepidoptera: Bombycidae) and comparison with other lepidopteran insects

The complete mitochondrial genome (mitogenome) of Bombyx mori strain Dazao (Lepidoptera: Bombycidae) was determined to be 15,653 bp, including 13 protein-coding genes (PCGs), two rRNA genes, 22 tRNA genes and a A + T-rich region. It has the typical gene organization and order of mitogenomes from lepidopteran insects. The AT skew of this mitogenome was slightly positive and the nucleotide composition was also biased toward A + T nucleotides (81.31%). All PCGs were initiated by ATN codons, except for cytochrome c oxidase subunit 1 (cox1) gene which was initiated by CGA. The cox1 and cox2 genes had incomplete stop codons consisting of just a T. All the tRNA genes displayed a typical clover-leaf structure of mitochondrial tRNA. The A + T-rich region of the mitogenome was 495 bp in length and consisted of several features common to the lepidopteras. Phylogenetic analysis showed that the B. mori Dazao was close to Bombycidae.     

The complete mitochondrial genome of the grand jackknife clam, Solen grandis (Bivalvia: Solenidae): a novel gene order and unusual non-coding region

Molluscs in general, and bivalves in particular, exhibit an extraordinary degree of mitochondrial gene order variation when compared with other metazoans. The complete mitochondrial genome of Solen grandis (Bivalvia: Solenidae) was determined using long-PCR and genome walking techniques. The entire mitochondrial genome sequence of S. grandis is 16,784 bp in length, and contains 36 genes including 12 protein-coding genes (atp8 is absent), 2 ribosomal RNAs, and 22 tRNAs. All genes are encoded on the same strand. Compared with other species, it bears a novel gene order. Besides these, we find a peculiar non-coding region of 435 bp with a microsatellite-like (TA)₁₂ element, poly-structures and many hairpin structures. In contrast to the available heterodont mitochondrial genomes from GenBank, the complete mtDNA of S. grandis has the shortest cox3 gene, and the longest atp6, nad4, nad5 genes.     

Characterization of the complete mitochondrial genome of Cryptotermes domesticus (Blattodea: Kalotermitidae): Genome description and phylogenetic implications

The complete mitochondrial genome of Cryptotermes domesticus (Haviland) was sequenced and annotated to study its characteristics and the phylogenetic relationship of C. domesticus to other termite species. The mitogenome of C. domesticus is a circular, close, and double-stranded molecule with a length of 15,655 bp. The sequenced mitogenome contains 37 typical genes, which are highly conserved in gene size, organization, and codon usage. Transfer RNA genes (tRNAs) also have typical secondary structures. All of the 13 protein-coding genes (PCGs) start with an ATN codon, except for nad4, which starts with GTG and terminates with the terminal codon TAA and TAG or the incomplete form T-- (cox2 and nad5). Most tRNAs have a typical cloverleaf structure, except for trnS1, in which this form is replaced by a simple loop and lacks the dihydrouridine (DHU) arm. The nucleotide diversity (Pi) and nonsynonymous (Ka)/synonymous (Ks) mutation rate ratios indicate that nad1, cox1, and cox3 are the most conserved genes, and that cox1 has the lowest rate of evolution. In addition, an 89 bp repeated sequence was found in the A + T-rich region. Phylogenetic analysis was performed using Bayesian inference (BI) and maximum likelihood (ML) methods based on 13 PCGs, and the monophyly of Kalotermitidae was supported.     

Characterization and Phylogenetic Analysis of the Mitochondrial Genome of Shiraia bambusicola Reveals Special Features in the Order of Pleosporales

Shiraia bambusicola P. Henn. is a pathogenic fungus of bamboo, and its fruiting bodies are regarded as folk medicine. We determined and analyzed its complete mitochondrial DNA sequence (circular DNA molecule of 39,030 bp, G + C content of 25.19%). It contains the typical genes encoding proteins involved in electron transport and coupled oxidative phosphorylation (nad1-6 and nad4L, cob and cox1-3), one ATP synthase subunit (atp6), 4 hypothetical proteins, and two genes for large and small rRNAs (rnl and rns). There is a set of 32 tRNA genes comprising all 20 amino acids, and these genes are evenly distributed on the two strands. Phylogenetic analyses based on concatenated mitochondrial proteins indicated that S. bambusicola clustered with members of the order Pleosporales, which is in agreement with previous results. The gene arrangements of Dothideomycetes species contained three regions of gene orders partitioned in their mitochondrial genomes, including block 1 (nad6-atp6), block 2 (nad1-cox3) and block 3 (genes around rns). S. bambusicola displayed unique special features that differed from the other Pleosporales species, especially in the coding regions around rns (trnR-trnY). Moreover, a comparison of gene orders in mitochondrial genomes from Pezizomycotina revealed that although all encoded regions are located on the same strand in most Pezizomycotina mtDNAs, genes from Dothideomycetes species had different orientations, as well as diverse positions and colocalization of genes (such as cox3, cox1-cox2 and nad2–nad3); these distinctions were regarded as class-specific features. Interestingly, two incomplete copies of the atp6 gene were found on different strands of the mitogenomic DNA, a finding that has not been observed in the other analyzed fungal species. In our study, mitochondrial genomes from Dothideomycetes species were comprehensively analyzed for the first time, including many species that have not appeared in previous reports.     

Detection of two different mitochondrial genomes in a gall wasp species,Andricus mairei (Hymenoptera: Cynipoidea: Cynipidae)

Mitochondrial genomes (mitogenomes) have been used widely in comparative and evolutionary genomics, molecular evolution, phylogenetics, and population genetics, but very limited information is available for the family Cynipidae. In this report, we described the mitogenome of Andricus mairei. The mitogenome of A. mairei was 16,514 bp in length and contained a typical set of 37 genes. Two control regions (CRs) were detected, one being a partial reverse repeat of the other. In a comparison with the putative ancestral mitogenome, gene rearrangements were found in transfer RNA (tRNA) genes, protein-coding genes and ribosomal RNA (rRNA) genes. Consistent with other Cynipidae species, the gene rearrangement of A. mairei had four obvious characteristics: trnE and trnF had inverted and swapped positions; rrnL and rrnS genes had moved into the cob–nad1 junction; a novel tRNA gene cluster trnL1–trnI–trnL2–trnW–trnM–trnQ had been formed between nad1 and nad2; and trnV had inverted and moved to the nad2–cox1 gene junction. Furthermore, A. mairei had two types of mitochondrial circular DNA molecules. Type II differed from type I in an inverted rearrangement of a large fragment of 3349 nucleotides, including two CRs and two rRNA genes.     

Complete mitochondrial genome of the darkling beetle Gonocephalum outreyi (Coleoptera: Tenebrionidae) with phylogenetic implications

The complete mitochondrial genome (mitogenome) of Gonocephalum outreyi was determined by using next-generation sequencing approach. The full length of this mitogenome is 15,836?bp, which consists of 37 typical metazoan mitochondrial genes with an identical genome organization to ancestral insects. The majority of the protein-coding genes begin with the codon ATN, except for cox1 and cox2 with AAT and AAA, respectively. To elucidate the phylogenetic position of G. outreyi, we used various sequence coding schemes for protein-coding genes and the combined nucleotide sequences of all mitochondrial genes for tree building under the Bayesian and Maximum Likelihood inferences. The phylogenetic results consistently supported G. outreyi as a member of the family Tenebrionidae. The monophyly of both Tenebrionoidea and Tenebrionidae were strongly supported. The Scraptiidae and Melandryidae were recovered to be non-monophyletic in regards to the Osphya. Within Tenebrionidae, the subfamilies Diaperinae and Tenebrioninae were found to be non-monophyletic.     

Performance of distance-based DNA barcoding in the molecular identification of Primates

For comparative primatology proper recognition of basal taxa (i.e. species) is indispensable, and in this the choice of a suitable gene with high phylogenetic resolution is crucial. For the goals of species identification in animals, the cytochrome c oxidase subunit 1 (cox1) has been introduced as standard marker. Making use of the difference in intra- and interspecific genetic variation – the DNA barcoding gap – cox1 can be used as a fast and accurate marker for the identification of animal species. For the Order Primates we compare the performance of cox1 (166 sequences; 50 nominal species) in species-identification with that of two other mitochondrial markers, 16S ribosomal RNA (412 sequences, 92 species) and cytochrome b (cob: 547 sequences, 72 species). A wide gap exist between intra- and interspecific divergences for both cox1 and cob genes whereas this gap is less apparent for 16S, indicating that rRNA genes are less suitable for species delimitation in DNA barcoding. For those species where multiple sequences are available there are significant differences in the intraspecific genetic distances between different mitochondrial markers, without, however, showing a consistent pattern. We conclude that cox1 allows accurate differentiation of species and as such DNA barcoding may have an important role to play in comparative primatology.     

The Mitochondrial Genome of Chlorogonium elongatum Inferred from the Complete Sequence

The 22,704-bp circular mitochondrial DNA (mtDNA) of the chlamydomonad alga Chlorogonium elongatum was completely cloned and sequenced. The genome encodes seven proteins of the respiratory electron transport chain, subunit 1 of the cytochrome oxidase complex (cox1), apocytochrome b (cob), five subunits of the NADH dehydrogenase complex (nad1, nad2, nad4, nad5, and nad6), a set of three tRNAs (Q, W, M), and the large (LSU)- and small (SSU)-subunit ribosomal RNAs. Six group-I introns were found, two each in the cox1, cob, and nad5 genes. In each intron an open reading frame (ORF) related to maturases or endonucleases was identified. Both the LSU and the SSU rRNA genes are split into fragments intermingled with each other and with other genes. Although the average A + T content is 62.2%, GC-rich clusters were detected in intergenic regions, in variable domains of the rRNA genes, and in introns and intron-encoded ORFs. A comparison of the genome maps reveals that C. elongatum and Chlamydomonas eugametos mtDNAs are more closely related to one another than either is to Chlamydomonas reinhardtii mtDNA. Received: 3 November 1997 / Accepted: 12 January 1998