The complete mitochondrial genome sequence of the nerippe fritillary butterfly, Argynnis nerippe, which is listed as an endangered species in Korea, is described with an emphasis on the A+T-rich region. The 15,140-bp long circular molecule consisted of 13 protein-coding genes, two rRNA genes, 22 tRNA genes and one control region, known in insect as the A+T-rich region, as found in typical metazoans. The 329-bp long A+T-rich region located between srRNA and tRNAMet possessed the highest A/T content (95.7%) than any other region of the genome. Along with the several conserved sequences found typically in the lepidopteran insects the genome contained one tRNAMet-like and tRNALeu(UUR) -like sequence in the A+T-rich region.

DNA ‘barcoding’ has potential applications in insect pest monitoring and quarantine since large numbers of DNA sequences for insect species identification have been reported in recent years. However, the exact number of relevant COI sequences in public databases such as NCBI (http://www.ncbi.nlm.nih.gov/) is not readily available. The IMGD (Insect Mitochondrial Genome Database; http://www.imgd.org) contains 162,847 partially sequenced mitochondrial gene entries originated from 35,067 hexapod species and makes it possible to check whether DNA sequences have been previously reported or not for certain insect species. In this study, we applied the IMGD to establish baseline data for the forest pest insects in Korea, before constructing a DNA barcode system. Retrieving data from the IMGD, we recognized that DNA sequences were already available for 73 of 259 species known as forest pest insects in Korea. Most of the 73 species with DNA sequences are common pests worldwide but there maining 186 species are endemic to the Eastern Palearctic region. Based on these data, we are proceeding to construct a Korean Forest Insect Pest DNA barcode database.

We have determined the complete mitochondrial genome of the yellow-spotted long horned beetle, Psacothea hilaris (Coleoptera: Cerambycidae), an endangered insect species in Korea. The 15,856-bp long P. hilaris mitogenome harbors gene content typical of the animal mitogenome and a gene arrangement identical to the most common type found in insect mitogenomes. As with all other sequenced coleopteran species, the 5-bp long TAGTA motif was also detected in the intergenic space sequence located between tRNASer (UCN) and ND1 of P. hilaris. The 1,190-bp long non-coding A+T-rich region harbors an unusual series of seven identical repeat sequences of 57-bp in length and several stretches of sequences with the potential to form stem-and-loop structures. Furthermore, it contains one tRNAArg-like sequence and one tRNALys-likes equence. Phylogenetic analysis among available coleopteran mitogenomes using the concatenated amino acid sequences of PCGs appear to support the sister group relationship of the suborder Polyphaga to all remaining suborders, including Adephaga, Myxophaga, and Archostemata. Among the two available infraorders in Polyphaga, a monophyletic Cucujiformia was confirmed, with the placement of Cleroidea as the basal lineage for Cucujiformia. On the other hand, the infraorder Elateriformia was not identified as monophyletic, thereby indicating that Scirtoidea and Buprestoidea are the basal lineages for Cucujiformia and the remaining Elateriformia.

The complete nucleotide sequences of the mitochondrial genome (mitogenome) from the white-spotted flower chafer, Protaetia brevitarsis (Coleoptera: Cetoniidae) was determined. The 20,319-bp long circular genome is the longest among the completely sequenced arthropods. This extraordinary length of the genome stemmed from 5,654-bp long A+T-rich region composed of twenty-eight 117-bp tandem repeats, seven 82-bp tandem repeats, and each two 19-bp and 38-bp tandem repeats. The P. brevitarsis contains a typical gene complement, order, and arrangement identical to most common type found in insects. The P. brevitarsis COI gene does not have typical ATN codon. Thus, we also designated it as AAC (asparagine), which is found in the start context of all sequenced Polyphaga within Coleoptera. All tRNAs showed stable canonical clover-leaf structure of other mt tRNAs, except for tRNASer (AGN), DHU arm of which could not form stable stem-loop structure. The 5bp-long motif sequence (TAGTA) that has been suggested to be the possible binding site for the transcription termination peptide for the major-strand also was found betweent RNASer (UCN) and ND1, as have been detected in all sequenced coleopteran insects.

Mitochondrial genome is inherited in maternal origin without recombination by mating and its specific regions have been used to monitor insect pest populations in agriculture. The oriental fruit moth, Grapholita molesta, is a serious pest on apple industry by its direct damage on fruits. This study reports a full sequence of mitochondrial genome of G. molesta. Sequence contigs were made by primary PCRs on conserved regions and subsequent PCRs to fill the gaps. Annotated genes were highly matched to the sequences of other lepidopteran species. However, a few positions of tRNA genes on the genome were different to other mitochondrial genomes.

The 15,338-bp long complete mitochondrial genome (mitogenome) of the Japanese oak silkmoth, Antheraeayamamai (Lepidoptera: Saturniidae) was determined. This genome has a gene arrangement identical to those of all other sequenced lepidopteran insects, but differs from the most common type, as the result of the movement of tRNAMet to a position 5’-upstream of tRNAIle. No typical start codon of the A. yamamai COI gene is available. Instead, a tetranucleotide, TTAG, which is found at the beginning context of all sequenced lepidopteran insects was tentatively designated as the start codon for A. yamamai COI gene. Three of the 13 protein-coding genes (PCGs) harbor the incomplete termination codon, T or TA. All tRNAs formed stable stem-and-loop structures, with the exception of tRNASer(AGN), the DHU arm of which formed a simple loop as has been observed in many other metazoan mt tRNASer(AGN). The 334-bp long A+T-rich region is noteworthy in that it harbors tRNA-likestructures, as has also been seen in the A+T-rich regions of other insect mitogenomes. Phylogenetic analyses of the available species of Bombycoidea, Pyraloidea, and Tortricidea bolstered the current morphology-based hypothesis that Bombycoidea and Pyraloidea are monophyletic (Obtectomera). As has been previously suggested, Bombycidae (Bombyxmori and B.mandarina) and Saturniidae (A.yamamai and Caligula boisduvalii) formed a reciprocal monophyletic group.

We determined the complete mitochondrial genome of the yellow-spotted long horned beetle, Psacothea hilaris (Coleoptera: Cerambycidae) that is an endangered insect species in Korea. The genome was sequenced from four overlapping fragments: two short fragments and two long fragments. The 15,857-bp long P. hilaris mitochondrial genome has the gene content typical of animal mitochondrial genome: 13 protein-coding genes, 22 tRNA genes, 2 ribosomal genes, and one non-coding A+T-rich region. The gene arrangement of the molecule is identical to the most common type found among insect mitochondrial gene arrangement that is regarded as ancestral for insects. Like several other coleopteran species the P. hilaris COI gene has typical mitochondrial start codon ATT. The 1,190-bp long A+T-rich region contains 57-bp long seven identical repeat sequences and at least seven stem-and-loop structures, composed of stems with perfect matches and loops with variable size. All P. hilaris tRNAs can be folded into the typical clover-leaf structure, with the exception of tRNASer(AGN), the dihydrouridine (DHU) arm of which forms a simple loop. After more genomic and phylogenetic analyses are performed, further detailed information will be available.

In this study, we determined the complete mitochondrial genome of the jewel beetle, Chrysochroa fulgidissima (Coleoptera: Buprestidae), from four overlapping fragments. The 15,592-bp long C. fulgidissima mitogenome exhibits a gene arrangement and content identical to the most common type in insects. The start codon of the C. fulgidissima COI gene is unusual, in that no typical ATN codon is available. The 875-bp A+T-rich region is the shortest among the coleopteran mitogenomes that have thus far been sequenced in their entirety. The most unusual feature of the genome is the presence of three tRNA-like sequences within the A+T-rich region: two tRNALeu(UUR)-like sequences and one tRNAAsnlike sequence. These sequence stretches evidence the proper anticodon sequence and the potential to form secondary structures, but also harbor many mismatches in the stems. Phylogenetic analysis using a concatenation of 13 amino acid sequences of protein-coding genes among the available sequenced species of coleopteran superfamilies (Buprestoidea and Elateroidea belonging to the infraorder Elateriformnia, and Chrysomeloidea and Tenebrioroidea belonging to the infraorder Cucujoiformia) by Bayesian inference, maximum-parsimony analyses, and maximum-likelihood analysis unexpectedly revealed a lack of support for monophyletic Elateriformia.

The 15,389-bp long complete mitochondrial genome (mitogenome) of the redspotted apollo butterfly, Parnassius bremeri (Lepidoptera: Papilionidae) was determined. This genome has a gene arrangement identical to those of all other sequenced lepidopteran insects, but differs from the most common type, as the result of the movement of tRNAMet to a position 5’-upstream of tRNAIle. No typical start codon of the P. bremeri COI gene is available. Instead, a tetranucleotide, TTAG, which is found at the beginning context of all sequenced lepidopteran insects was tentatively designated as the start codon for P. bremeri COI gene. All protein-coding genes (PCGs), but COII (T) have complete termination codon TAA or TAG. One of the most unusual feature of the P. bremeri mitochondrial genome is the presence of two tRNA-like structures, such as the tRNATrp-like sequence and tRNALeu (UUR)-like sequence with proper anticodon and clover-leaf structures in the 514-bp long A+T-rich region. Furthermore, the A+T-rich region possesses three sequences that have the potential to form stem-and-loop structures, flanked by the conserved sequences, "TA(A)TA" at the 5’ end and "G(A)nT’ at the 3’ end. After more genomic and phylogenetic analyses are performed, further detailed information will be available.

The completely sequenced mitochondrial genome of the brown marmorated stink bug, Halyomorpha halys, is a circular molecule of 16,518 bp with a total A+T content of 76.4%. Nucleotide composition and codon usage of this genome are near the means observed in other 12 hemipteran mitochondrial genomes; however, the initiation codon for CO1 gene appears to be TTG, dissimilar to what has been seen in the 12 mitochondrial genomes. In this genome, the A+T rich region between srRNA and tRNAIle gene includes two extensive repeat regions, in which each region includes 4 and 12 tandem repeats of a 73 bp sequence, respectively. The gene content, order, and structure of the H. halys mitochondrial genome are consistent with that of Triatoma dimidiata, belong to the same suborder Heteroptera, but different from two suborders, Auchenorrhynca and Sternorrhyncha, including various gene rearrangements. Analyzing phylogenetic relationship and comparing gene order and content of the 13 hemipteran mitochondrial genomes of three suborders, Heteroptera, Auchenorrhynca, and Sternorrhyncha, supported the morphology-based current hypothesis that both Auchenorrhynca and Sternorrhyncha are a monophyletic group.

Mitochondria are essential organelles of eukaryotic cells and plant cells contain varying numbers of mitochondrial genome sequences. Sizes and shapes of mitochondria differ within a tissue or in the same cells. Previously sequenced complete mitochondrial genome (NC_016118) of Brassica oleracea size was 360,271 bp, where segmental duplication (repeat block) was 141,800 bp. In this study, we resequenced this whole mitochondrial genome by using WGS (whole genome sequencing) and assembled organelles genome method (unpublished). Newly sequenced mitochondrial genome length was 219,975 bp and circle form. A new sequence segment of approximately 4,800 bp was obtained compared to the previous genome sequence without any large repeat block. Newly obtained mitochondria genome sequence was compared with recently reported mitochondria genome sequences of various species (B. oleracea, B. juncea, B. rapa, B. napus and B. carinata) and subspecies (cabbage, cauliflower, brussels sprouts, kohlrabi, broccoli and kale) by PCR using primers specifying different region of genome sequences. PCR analysis results have also confirmed the variation between previous and newly sequenced mitochondrial genome circles form. Thus, the results suggest new B. oleracea mitotype, including evolutionary events such as inheritance, rearrangement, genome compaction, and diversity