The complete mitochondrial genome of the oriental fruit moth <Emphasis Type="Italic">Grapholita molesta</Emphasis> (Busck) (Lepidoptera: Tortricidae)

The oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae) currently is one of the economically most destructive pest species of stone and pome fruits worldwide. Here we sequenced the complete mitochondrial genome of this pest. This genome is 15,776 bp long, with an A + T content of 81.24%, containing 37 typical animal mitochondrial genes and an A + T-rich region. All gene are arranged as hypothesized ancestral gene order of insects except for trnM, which was shuffled from 3′ downstream of trnQ to 5′ upstream of trnI. cox1 gene uses unusual CGA start codon, as that in all other sequenced lepidopteran mitochondrial genome. The secondary structures for the two rRNA genes were predicted. All helices typically present in insect mitochondrial rRNA genes are generated. A microsatellite sequence was inserted into the region of H2347 in rrnL in G. molesta and two other sequenced tortricid mitochondrial genomes, indicating that the insertion event in this helix might occurred anciently in family Tortricidae. All of the 22 typical animal tRNA genes have a typical cloverleaf structure except for trnS2, in which the D-stem pairings in the DHU arm are absent. An intergenic sequence is present between trnQ and nad2 as well as in other sequenced lepidopteran mitochondrial genomes, which was presumed to be a remnant of trnM gene and its boundary sequences after the duplication of trnM to the upstream of trnI in Lepidoptera. The A + T-rich region is 836 bp, containing six repeat sequences of “TTATTATTATTATTAAATA(G)TTT.”     

The complete mitochondrial genome of the butterfly <Emphasis Type="Italic">Apatura metis</Emphasis> (Lepidoptera: Nymphalidae)

As an important pest in the Slender Leaved Willow (Salix alba), Apatura metis is called Freyer’s purple emperor, and its mitochondrial genome is 15,236 bp long. The encoded genes for 22 tRNA genes, two ribosomal RNA (rrnL and rrnS) genes, and 13 protein-coding genes (PCGs), and a control region in the A. metis mitochondria are highly homologous to other lepidopteran species. The mitochondrial genome of A. metis is biased toward a high A + T content (A + T = 80.5%). All protein-coding genes, except for COI begins with the CGA codon as observed in other lepidopterans, start with a typical ATN initiation codon. All tRNAs show the classic clover-leaf structure, except that the dihydrouridine (DHU) arm of tRNA ^Ser(AGN) forms a simple loop. The A. metis A + T-rich region contains some conserved structures including a structure combining the motif ‘ATAGA’ and 19 bp poly (T) stretch, which is similar to those found in other lepidopteran mitogenomes. The phylogenetic analyses of lepidopterans based on mitogenomes sequences demonstrate that each of the six superfamilies is monophyletic, and the relationship among them is (((Noctuoidea + (Geometroidea + Bombycoidea)) + Pyraloidea) + Papilionoidea) + Tortricoidea. In Papilionoidea group, our conclusion argues that ((Lycaenidae + Pieridae) + Nymphalidae) + Papilionidae.     

The complete mitochondrial genome of <Emphasis Type="Italic">Thrinchus schrenkii</Emphasis> (Orthoptera: Caelifera,Acridoidea, Pamphagidae)

Complete nucleotide sequences of mitochondrial genome (mitogenome) of Thrinchus schrenkii (Orthoptera: Acridoidea: Pamphagidae) were determined. It is 15672 bp in length and contains 71.2% A + T. All T. schrenkii protein-coding sequences except for the cytochrome oxidase subunit I (COI) start with a typical ATN codon. Instead, CCG, which is a rare but possible initiation codon, is located at the initiation context of COI. The usual termination codons (TAA and TAG) were found from 12 PCGs. However, the ND5 had incomplete termination codon (T). All tRNA genes could be folded into the typical cloverleaf secondary structure, excluding tRNA ^Ser(AGN) which forms another structure according to the Steinberg–Cedergren tertiary structure. The sizes of the large and small ribosomal RNA genes are 1319 and 848 bp, respectively. The A + T content of the A + T-rich region is 78.7%, which is the lowest among the known mitogenome of Acridoidea.     

The complete mitochondrial genome of <Emphasis Type="Italic">Deracantha onos</Emphasis> (Orthoptera: Bradyporidae)

The complete mitochondrial genome 15,650 bp in size of the Deracantha onos has been determined. The gene content, base composition and codon usage of D. onos are coincident to typical hexapods mitochondrial genomes. Genes arrangement of D. onos is identical to Gryllotalpa orientalis, Ruspolia dubia and Anabrus simplex, in that the relative locations of tRNA^Lys and tRNA^Asp was different to that of Locusta migratoria. All tRNAs could be folded into the typical cloverleaf secondary structure, excluding tRNA^Ser(AGN) which forms another structure according to the Steinberg–Cedergren tertiary structure. Sequence analysis of the A + T-rich region with Dot-plot did not find any conspicuous repeat clusters. Two poly-thymine (poly-T) nucleotide stretches of 20 bp and 11 bp in size, which may involved in the recognition of replication origin, were found on the H-strand and L-strand in the A + T-rich region of the D. onos mitogenome, respectively. One open reading frame (ORF) 87 amino acids in size was found on the H-strand, but Protein Blast searches analysis indicated that it was a nonfunctional ORF.     

The mitochondrial genome of <Emphasis Type="Italic">Protohermes concolorus</Emphasis> Yang et Yang 1988 (Insecta: Megaloptera: Corydalidae)

The first complete mitochondrial genome of dobsonfly Protohermes concolorus Yang et Yang, 1988 (Megaloptera: Corydalidae) was sequenced in this study. The genome was a circular molecule of 15,851 bp containing the typical 37 genes that arranged in the same order as that of the putative ancestor of hexapods. Sequences overlaps were observed between several neighbor genes, which made the genome relatively compact. The tRNA-Ser (GCT) could not be folded into typical secondary structure because its DHU arm was replaced with a simple loop. Six of the 13 protein genes were terminated with a single T adjacent to a downstream tRNA gene in the same strand. The variation of GC content caused the different nucleotide substitution patterns of the protein genes. The genome was AT-biased with a total A + T content of 75.83% which was also demonstrated by the codon usage. The control region was the most AT-rich region with a sub-region of even higher A + T content. Protein genes of two strands presented opposite CG-skew trends which was also reflected by the codon usage. For most of the amino acids, the protein coding sequences did not prefer to use the cognate codons of corresponding tRNAs and the codon usage of the protein genes was not random. The variation of nucleotide substitution patterns of protein genes was significantly correlated with the GC content. The phylogenetic analyses based on all the 13 protein genes showed that Megaloptera was the sister group of other holometabolous insects except Coleoptera.     

The complete nucleotide sequence of the mitochondrial genome of <Emphasis Type="Italic">Phthonandria atrilineata</Emphasis> (Lepidoptera: Geometridae)

Using long-polymerase chain reaction (Long-PCR) method, we determined the complete nucleotide sequence of the mitochondrial genome (mitogenome) of Phthonandria atrilineata. The complete mtDNA from P. atrilineata was 15,499 base pairs in length and contained 13 protein-coding genes (PCGs), 2 rRNA genes, 22 tRNA genes, and a control region. The P. atrilineata genes were in the same order and orientation as the completely sequenced mitogenomes of other lepidopteran species. The nucleotide composition of P. atrilineata mitogenome was biased toward A + T nucleotides (81.02%), and the 13 PCGs show different A + T contents that range from 73.25% (cox1) to 92.12% (atp8). Phthonandria had the canonical set of 22 tRNA genes, that fold in the typical cloverleaf structure described for metazoan mt tRNAs, with the unique exception of trnS(AGN). The phylogenetic relationships were reconstructed with the concatenated sequences of the 13 PCGs of the mitochondrial genome, which confirmed that P. atrilineata is most closely related to the superfamily Bombycoidea.     

The complete mitogenome and phylogenetic analysis of <Emphasis Type="Italic">Bombyx mandarina</Emphasis> strain Qingzhou

The complete nucleotide sequence of the mitogenome of Bombyx mandarina strain Qingzhou was determined. The circular genome is 15,717 bp long and has the typical gene organization and order of lepidopteran mitogenomes. All protein-coding sequences are initiated with a typical ATN codon, except the COI gene, which has a 4-bp TTAG putative initiator codon. Eleven of the 13 protein-coding gene have a complete termination codon (all TAA), but the remaining two genes terminate with incomplete codons. All transfer RNAs (tRNAs) have a clover-leaf structure typical of the mitochondrial tRNAs, and some of them have a mismatch in the four-stem-and-loop structure. The length of the A + T rich region of B. mandarina strain Qingzhou is 495 bp, shorter than that of B. mandarina strain Tsukuba (747 bp) but similar to that of Bombyx mori. Phylogenetic analysis based on the whole mitochondrial genome sequences of the available sequenced species (B. mori strains C-108, Aojuku, Backokjam, and Xiafang, B. mandarina strains Tsukuba, Ankang, and Qingzhou, and Antheraea pernyi) shows the origin of the domesticated silkmoth B. mori to be the Chinese B. mandarina. Nuclear mitochondrial pseudogene sequences were detected in the nuclear genome of B. mori with the MEGA BLAST search program. A phylogenetic analysis of these nuclear mitochondrial pseudogene sequences suggests that B. mori was domesticated independently in different areas and periods.     

The complete mitochondrial genome of <Emphasis Type="Italic">Spilonota lechriaspis</Emphasis> Meyrick (Lepidoptera: Tortricidae)

We determined the nucleotide sequence of the mitochondrial genome (mtgenome) of Spilonota lechriaspis Meyrick (Lepidoptera: Tortricidae). The entire closed circular molecule is 15,368 bp and contains 37 genes with the typical gene complement and order for lepidopteran mtgenomes. All tRNAs except tRNA^Ser(AGN) can be folded into the typical cloverleaf secondary structures. The protein-coding genes (PCGs) have typical mitochondrial start codons, with the exception of COI, which uses the unusual CGA one as is found in all other Lepidoptera sequenced to date. In addition, six of 13 PCGs harbor the incomplete termination codons, a single T. The A + T-rich region contains some conserved structures that are similar to those found in other lepidopteran mtgenomes, including a structure combining the motif ‘ATAGA’, a 19-bp poly(T) stretch and three microsatellite (AT)_n elements which are part of larger 122+ bp macrorepeats. This is the first report of macrorepeats in a lepidopteran mtgenome.     

The complete mitochondrial genome of the cockroach <Emphasis Type="Italic">Eupolyphaga sinensis</Emphasis> (Blattaria: Polyphagidae) and the phylogenetic relationships within the Dictyoptera

We present the complete mitochondrial DNA sequence of Eupolyphaga sinensis. This closed circular molecule is 15553 bp long and consists of 37 genes that encode for 13 inner membrane proteins, 2 ribosomal RNAs and 22 transfer RNAs. The genome shares the gene order and orientation with previously known Blattaria mitochondrial genomes. All tRNAs could be folded into the typical cloverleaf secondary structure, but the tRNASer (AGN) appears to be missing the DHU arm. The A + T-rich region is 857 bp long and longer than other cockroaches. Based on the concatenated amino acid sequences of all protein coding genes of E. sinensis in conjunction with those 23 other arthropod sequences, we reconstruct the phylogenetic tree. Phylogenetic analyses shows that Blataria (including Isoptera) and the Mantodea are sister groups. Furthermore the relationship of the three basal clades of winged insects are different from the three previous hypotheses ((Ephemeroptera + Odonata) +Neoptera, Ephemeroptera + (Odonata + Neoptera), Odonata + (Ephemeroptera +Neoptera)). The Ephemeroptera (Parafronurus youi) clusters with the Plecoptera (Pteronarcys princes).     

<Emphasis Type="Italic">Johnius</Emphasis> (<Emphasis Type="Italic">Johnius</Emphasis>) <Emphasis Type="Italic">majan</Emphasis> sp. nov., a sciaenid fish (Pisces: Sciaenidae) from Oman,Indian Ocean

Johnius (Johnius) majan sp. nov. is described on the basis of 8 specimens (117–158 mm in standard length) from Oman, Indian Ocean. The new species is distinguished from its congeners by the following combination of characters: black axillary spot on upper pectoral fin base; dorsal soft rays 29–32; anal soft rays 8; scales above lateral line 6, below 11; eye diameter 22.9–28.9% HL; interorbital width 32.0–38.0% HL; gill rakers 5–6 + 15–18 = 21–24; no mental barbel; last well developed pleural rib on 7th vertebra; swim bladder appendages 11; vertebrae 10 + 14 = 24.     

The complete mitochondrial genome structure of snow leopard <Emphasis Type="Italic">Panthera uncia</Emphasis>

The complete mitochondrial genome (mtDNA) of snow leopard Panthera uncia was obtained by using the polymerase chain reaction (PCR) technique based on the PCR fragments of 30 primers we designed. The entire mtDNA sequence was 16 773 base pairs (bp) in length, and the base composition was: A—5,357 bp (31.9%); C—4,444 bp (26.5%); G—2,428 bp (14.5%); T—4,544 bp (27.1%). The structural characteristics [0] of the P. uncia mitochondrial genome were highly similar to these of Felis catus, Acinonyx jubatus, Neofelis nebulosa and other mammals. However, we found several distinctive features of the mitochondrial genome of Panthera unica. First, the termination codon of COIII was TAA, which differed from those of F. catus, A. jubatus and N. nebulosa. Second, tRNA^{Ser (AGY)}, which lacked the ‘‘DHU’’ arm, could not be folded into the typical cloverleaf-shaped structure. Third, in the control region, a long repetitive sequence in RS-2 (32 bp) region was found with 2 repeats while one short repetitive segment (9 bp) was found with 15 repeats in the RS-3 region. We performed phylogenetic analysis based on a 3 816 bp concatenated sequence of 12S rRNA, 16S rRNA, ND2, ND4, ND5, Cyt b and ATP8 for P. uncia and other related species, the result indicated that P. uncia and P. leo were the sister species, which was different from the previous findings.     

The complete mitochondrial genome analysis of the tiger (<Emphasis Type="Italic">Panthera tigris</Emphasis>)

The complete mitochondrial genomes of five tiger samples from three subspecies (P. t. sumatrae, P. t. altica, and P. t. tigris) were successfully obtained by using 26 specifically designed Panthera-specific primer sets. The genome organization and gene arrangement of the five tiger samples were similar to each other; however polymorphic tandem repeat sequences were observed in the control region (CR). This led to a difference in the genome lengths obtained from these five samples with an average size of 16,994 bp for the five tiger mitochondrial genomes. The nucleotide base composition was on average as follows: A, 31.8%; T, 27.0%; C, 26.6%; G, 14.6% and exhibited compositional asymmetry. Most of tiger mitochondrial genome characteristics are similar to those of other common vertebrate species; however, some distinctive features were observed in the CR. First, the repetitive sequence 2 (RS 2) contained two repeat units of 80 bp and the first 15 bp of what would be the third repeat motif. The repetitive sequence 3 (RS 3) contained 47–50 repeat motifs of a shorter 8 bp (ACGTAYAC)_n. Second, length heteroplasmy polycystosine (poly-C) stretches was observed at the end of the HV I locus in all tiger samples.     

The Discriminatory Transfer Routes of tRNA Genes Among Organellar and Nuclear Genomes in Flowering Plants: A Genome-Wide Investigation of <Emphasis Type="Italic">indica</Emphasis> Rice

The transfer and integration of tRNA genes from organellar genomes to the nuclear genome and between organellar genomes occur extensively in flowering plants. The routes of the genetic materials flowing from one genome to another are biased, limited largely by compatibility of DNA replication and repair systems differing among the organelles and nucleus. After thoroughly surveying the tRNA gene transfer among organellar genomes and the nuclear genome of a domesticated rice (Oryza sativa L. ssp. indica), we found that (i) 15 mitochondrial tRNA genes originate from the plastid; (ii) 43 and 80 nuclear tRNA genes are mitochondrion-like and plastid-like, respectively; and (iii) 32 nuclear tRNA genes have both mitochondrial and plastid counterparts. Besides the native (or genuine) tRNA gene sets, the nuclear genome contains organelle-like tRNA genes that make up a complete set of tRNA species capable of transferring all amino acids. More than 97% of these organelle-like nuclear tRNA genes flank organelle-like sequences over 20 bp. Nearly 40% of them colocalize with two or more other organelle-like tRNA genes. Twelve of the 15 plastid-like mitochondrial tRNA genes possess 5′- and 3′-flanking sequences over 20 bp, and they are highly similar to their plastid counterparts. Phylogenetic analyses of the migrated tRNA genes and their original copies suggest that intergenomic tRNA gene transfer is an ongoing process with noticeable discriminatory routes among genomes in flowering plants. Electronic Supplementary Material Electronic Supplementary material is available for this article at and accessible for authorised users. Reviewing Editor: Dr. David Guttman     

The complete mitochondrial genome of the leafminer <Emphasis Type="Italic">Liriomyza trifolii</Emphasis> (Diptera: Agromyzidae)

Liriomyza trifolii (Diptera: Agromyzidae) is one of the most economically significant pests in the world. In this paper we present sequence data for the complete mitochondrial genome of L. trifolii. The circular genome is 16,141 bp long and contains one encoding region including 37 genes and one non-coding A+T-rich region. Gene numbers and organization is similar to that of the typical insect mitochondrial genomes except that two additional tRNA genes are found in the A+T-rich region (tRNA^Thr and tRNA^Leu(UUR)). All of the protein initiation codons are ATN, except ND1 which begins with GTG and COI which is initiated by the quadruplet ATCA. The 22 tRNA anticodons of L. trifolii match those observed in Drosophila yakuba, and all of tRNAs form the typical cloverleaf structure except for tRNA^Ser(AGN), which has lost the DHU-arm. The A+T-rich region of L. trifolii also contains two previously noted Diperan features—a highly conserved polyT stretch and a (TA)_n stretch.     

Identification of <Emphasis Type="Italic">Elymus</Emphasis> (Triticeae,Poaceae) and its related genera genomes by RFLP analysis of PCR-amplified <Emphasis Type="Italic">Adh</Emphasis> genes

Elymus L. is the largest genus in Triticeae, containing about 150 species with four recognized genome donors (St, H, P, and W). Traditionally, the genome compound of this genus is identified based on cytological data. Recently, molecular phylogenetic analysis was used to investigate its genomic combination. Here we describe a restriction fragment length polymorphism (RFLP) assay based on digesting alcohol dehydrogenase (Adh) amplicons with two restriction enzyme combinations, EcoRI–HindIII and EcoRI–PstI, which easily can be used to distinguish Elymus and its closely related genera genomes. The method includes only four steps: (1) amplifying nuclear Adh genes with universal primers; (2) purifying and cloning PCR products; (3) digesting plasmids with restriction enzymes that identify a given genome; (4) running the digested products on an agarose gel and identify the sample based on the restriction profiles. Results showed that: (1) PCR products ranged from 1,200 to 2,000 bp; (2) Adh2 gene was amplified from all the tested genomes; Adh1 gene was amplified from almost all of the tested genomes except the W genome; Adh3 gene was amplified only from the St genome; (3) the EcoRI–HindIII combination was effective to distinguish different Adh gene types (Adh1, Adh2, and Adh3); (4) the Adh2–EcoRI–PstI fragments could be used to distinguish Elymus and its closely related genera genomes. Therefore, This RFLP assay provides an inexpensive and simple means of identifying Elymus genomes.     

Mitochondrial genome sequence of <Emphasis Type="Italic">Unionicola parkeri</Emphasis> (Acari: Trombidiformes: Unionicolidae): molecular synapomorphies between closely-related <Emphasis Type="Italic">Unionicola</Emphasis> gill mites

The mitochondrial genome of Unionicola parkeri is a 14,734 bp circular DNA molecule. The sequence and annotation revealed a unique gene order, related to but distinct from the gene order in the closely related species U. foili. Mitochondrial tRNA sequences annotated in this genome predict non-canonical secondary structures for these molecules. The continuing patterns of unique gene orders and unusual tRNA structures in the Trombidiformes in general and Unionicola in particular support the use of phylogenetic approaches that use these types of molecular features as shared, derived character states. Further progress in using these molecular character states to reconstruct phylogeny will depend on careful annotation, especially of tRNA genes.     

Genetic diversity of <Emphasis Type="Italic">Gracilaria changii</Emphasis> (Gracilariaceae,Rhodophyta) from west coast,Peninsular Malaysia based on mitochondrial <Emphasis Type="Italic">cox</Emphasis>1 gene analysis

Mitochondrial cytochrome c oxidase subunit I (cox1) was employed to investigate the intraspecific genetic diversity of Gracilaria changii collected from various localities distributed along the west coast of Peninsular Malaysia. Gracilaria changii is an agarophyte with potential for commercialization in Malaysia as it has high yields of good quality agar with high gel strength for the production of food grade agar and agarose. The phylogeographic aspect of G. changii has not been studied despite its abundance and potential commercialization. In this study, six mitochondrial haplotypes (C1–C6) were revealed from 62 specimens varying by 0–3 bp over 923 bp. Results indicate that haplotype C1 is the common ancestor and the most widespread haplotype due to its prevalence in Morib, Gua Tanah, Middle Banks, Batu Besar, Batu Tengah, Sungai Pulai, and Kuala Sungai Merbok. In this study, Morib was suggested as contributing the highest intra-population diversity with the identification of three haplotypes. The mitochondrial marker cox1 is a highly divergent mitochondrial marker and is applicable for studies on species identification and assessment of genetic diversity of G. changii.     

Mitochondrial genome sequence of <Emphasis Type="Italic">Unionicola foili</Emphasis> (Acari: Unionicolidae): a unique gene order with implications for phylogenetic inference

The mitochondrial genome of Unionicola foili is circular, 14,738 bp in length, and contains several notable features. The sequence and annotation revealed a unique gene order, continuing a pattern of highly rearranged mitochondrial genomes in the Trombidiformes. U. foili mitochondrial tRNA sequences predict non-canonical secondary structures for these molecules, and our annotation suggests an in-frame fusion between the nad4L and nad5 genes in this genome. The unique gene order and unusual tRNA structures could serve as idiosyncratic characters and have the potential to be phylogenetically informative.     

Acaricidal effects of <Emphasis Type="Italic">Corymbia citriodora</Emphasis> oil containing <Emphasis Type="Italic">para</Emphasis>-menthane-3,8-diol against nymphs of <Emphasis Type="Italic">Ixodes ricinus</Emphasis> (Acari: Ixodidae)

The toxicity of para-menthane-3,8-diol (PMD), the main arthropod-repellent compound in the oil of the lemon eucalyptus, Corymbia citriodora, was evaluated against nymphs of Ixodes ricinus using five methods (A–E) of a contact toxicity bioassay. Mortality rates were estimated by recording numbers of dead nymphs at 30 min intervals during the first 5 h after the start of exposure and at longer intervals thereafter. The mortality rate increased with increasing concentration of PMD and duration of exposure with a distinct effect after 3.5 h. From the results obtained by methods A, C and E, the LC₅₀ range was 0.035–0.037 mg PMD/cm² and the LC₉₅ range was 0.095–0.097 mg PMD/cm² at 4 h of exposure; the LT₅₀ range was 2.1–2.8 h and the LT₉₅ range was 3.9–4.2 h at 0.1 mg PMD/cm². To determine the duration of toxic activity of PMD, different concentrations (0.002, 0.01, 0.1 mg PMD/cm²) were tested and mortality was recorded at each concentration after 1 h; thereafter new ticks were tested. This test revealed that the lethal activity of PMD remained for 24 h but appeared absent after 48 h. The overall results show that PMD is toxic to nymphs of I. ricinus and may be useful for tick control.