Eumenis autonoe, a member of the lepidopteran family Nymphalidae (superfamily Papilionoidea) is an endangered species, and is found only on one isolated remote island, Jeju in South Korea, on Halla Mt., at altitudes higher than 1,400 meters. In this study, we report the complete mitochondrial genome (mitogenome) of E. autonoe. The 15,489-bp long E. autonoe genome evidenced the typical gene content found in animal mitogenomes, and harbors the gene arrangement identical to all other sequenced lepidopteran insects, which differs from the most common type found in insects, due to the movement of tRNAMet to a position 5’-upstream of tRNAIle. As has been observed in many other lepidopteran insects, no typical ATN codon for the COI gene is available. Thus, we also designated the CGA (arginine) found at the beginning of the COI gene as a lepidopteran COI starter, in accordance with previous suggestions. The 678-bp long A+T-rich region, which is second longest in sequenced lepidopteran insects, harbors 10 identical 27-bp long tandem repeats plus one 13-bp long incomplete final repeat. Such a repeat sequence has been, thus far, only rarely detected in lepidopteran mitogenomes. The E. autonoe A+T-rich region harbors a poly-T stretch of 19 bp and a conserved ATAGA motif located at the end of the region, which have been suggested to function as structural signals for minor-strand mtDNA replication.

Eumenis autonoe belonging to a lepidopteran family Nymphalidae (superfamily Papilionoidea) is an endangered species in Korea. Historically, the species was distributed in Europe and Asian region including a wide region in Korean peninsula. However, in Korean peninsula, the species is found only in two isolate dregions: South in a remote island Jeju, where altitude is higher than1, 400 meter on Halla Mt. and North in far northern Korean peninsula around Mt. Bekdu. In this study, we report the complete mitochondrial genome of the endangered E. autonoe collected from Mt. Halla. The 15,489-bp long E. autonoe genome has a typical gene content found in animal mitochondrial genomes and contains the gene arrangement identical to all other sequenced lepidopteran insects, which differs from the most common type found in insects, as the result of the movement of tRNAMet to a position 5’-upstream of tRNAIle. As seen in many other lepidopteran insects, no typical ATN codon for COI gene is available. Thus, we tentatively designated the CGA (arginine) found at the beginning of the COI gene, as has been suggested for lepidopteran COI starter. The intergenic spacer sequence located between tRNASer (UCN) and ND1 of E. autonoe mitogenome also contains the ATACTAA motif which is conserved in all sequenced lepidopteran species. The 678-bp long A+T-rich region, which is longest in sequenced lepidopteran insects contains ten identical tandem repeats composed of 27 bp plus one 13-bp long identical incomplete final repeat. Such repeat sequence is rare in the lepidopteran mitogenomes known so far. The E. autonoe A+T-rich region also contains a poly-T stretch located at the end of the region as 19 bp and also contains the downstream conserved motif ATAGA that were previously suggested to serve as a structural signal for minor-strand mtDNA replication. Phylogenetic analysis using the concatenated 13 amino acid sequences of PCGs among available six lepidopteran superfamilies (Tortricoidea, Pyraloidea, Papilionoidea, Bombycoidea, Geometroidea, and Noctuoidea) rooted with three dipteran species with BI and ML analyses supported the following topology: ((((Bombycoidea + Geometroidea +Noctuoidea) + Papilionoidea) + Pyraloidea) + Tortricoidea). Within Papilionoidea, a closer relationship between Lycaenidae and Pieridae, excluding Nymphalidae was observed. Further fruitful information will be available after more analysis is done.

The 15,389-bp long complete mitogenome of the endangered red-spotted apollo butterfly, Parnassius bremeri (Lepidoptera: Papilionidae) was determined. This genome has a gene arrangement identical to those of all other sequenced lepidopteran insects, which have the gene order of tRNAMet, tRNAIle, and tRNAGln at the beginning. Due to the uncertainty the start codon for COI gene in insect has been discussed extensively. We propose the CGA sequence as the start codon for COI gene in lepidopteran insects, based on complete mitogenome sequences of lepidopteran insects including our P. bremerii and additional sequences of the COI start region from a diverse taxonomic range of lepidopteran species (a total of 51 species belonging to 15 families). As has been suggested in other sequenced lepidopteran insects the 18 bp-long poly-T stretch and the downstream conserved motif ATAGA that were previously suggested to serve as a structural signal for minor-strand mtDNA replication also was found at the 3’-end region of the P. bremerii A+T-rich region. In an extensive search to find out tRNA-like structure in the A+T-rich region, each one tRNATrp-like sequence and tRNALeu (UUR)-like sequence were found in the P. bremeri A+T-rich region, and most of other sequenced lepidopteran insects were shown to have tRNA-like structure within the A+T-rich region, thereby indicating that such feature is frequent in the lepidopteran A+T-rich region. Phylogenetic analysis using the concatenated 13 amino acid sequences and nucleotide sequences of PCGs of the four macrolepidopteran suferfamilies together with Tortricoidea and Pyraloidea well recovered a monophyly of Papilionoidea and a monophyly of Bombycoidea. However, Geometroidea and Noctuoidea were unexpectedly clustered as one group and placed this group to the sister group to Bombycoidea, instead of Papilionoidea in most analyses.

We determined the complete mitochondrial genome (mitogenome) of the Japanese Oak Silkmoth, Antheraea yamamai (Lepidoptera: Saturniidae) from two overlapping fragments and subsequent shotgun sequencing. The 15,601-bp long A. yamamai mitogenome contains gene arrangement and content identical to the most common arrangement found in lepidopteran insects. Most individual A. yamamai mitochondrial (mt) genes were well within the range found in the respective genes of other insects, except for small ribosomal RNA (1,037 bp). The 336-bp A+T-rich region is relatively smaller than that of other lepidopteran insects. The region is interesting in that it contains tRNA-like structures as found in the A+T-rich regions of other insect mitogenomes. The start codon of A. yamamai COI gene is unusual in that no typical one (ATN) is available. Three of the 13 protein-coding genes have incomplete termination codon T or TA. All tRNA formed stable stem-and-loop structure, except for tRNASer(AGN), the DHU arm of which formed a simple loop as seen in many other metazoan mt tRNASer(AGN).

We determined the complete mitochondrial genome (mitogenome) of the jewel beetle, Chrysochroa fulgidissima (Coleoptera: Buprestidae) from two overlapping fragments and subsequent sub fragments. The 15,592-bp long C. fulgidissima mitogenome contains gene arrangement and content identical to the most common arrangement found in insects. Most individual C. fulgidissima mitochondrial (mt) genes were well within the range found in the respective genes of other insects. The 875-bp A+T-rich region is shortest among the coleopteran mitogenomes sequenced in their entirety. The region is interesting in that it contains several stem-and-loop structures and tRNA-like structure found in the A+T-rich regions of other insect mitogenomes. As seen in other insect motogenomes the start codon of C. fulgidissima COI gene also is unusual because no typical start codon is available. Three of the 13 protein-coding genes have incomplete termination codon T or TA. All tRNA formed stable stem-and-loop structure, except for tRNASer(AGN), the DHU arm of which formed a simple loop as seen in many other metazoan mt tRNASer(AGN).