The Acoptolabrus changeonleei Ishikawa et Kim, 1983 (Coleoptera: Carabidae), has been listed as an endangered insect in South Korea. The complete mitochondrial genome of the species was 16,831 bp with a typical set of genes (13 protein-coding genes [PCGs], 2 rRNA genes, and 22 tRNA genes) and one non-coding region, with the arrangement identical to that observed in most insect genomes. Phylogenetic analyses with concatenated sequences of the 13 PCGs and 2 rRNA genes, using the Bayesian inference (BI) and maximum-likelihood (ML) methods, placed A. changeonleei as a sister to the within-subfamilial species Damaster mirabilissimus in Carabinae, with the highest nodal support by both analyses.

In this study, we developed 12 microsatellite markers specific to N. pygmaea using Illumina paired-end sequencing. Forty individuals of N. pygmaea collected from three currently known localities in South Korea were genotyped to validate these markers and to preliminarily assess population genetic characteristics. No locus showed significant deviation from the Hardy–Weinberg equilibrium (HWE). Our preliminary data indicate an absence of inbreeding in all populations and an absence of obvious genetic difference. The microsatellite markers developed in this study will be useful for studying the population genetics of N. pygmaea collected from other regions, including additional sites in South Korea.

Previously, a partial mitochondrial COI and CytB gene sequences have been used to infer genetic diversity and gene flow of the species. In this study, we additionally collected C. tripartitus from one previous and two new localities, sequenced the COI and CytB genes. Sequence divergence of current samples showed slightly lower values, but nearly equivalent to previous study. Haplotype diversity was still high and nucleotide diversity indicating that Seogwi-dong showed the highest estimates in both gene sequences. Gene flow among populations is high, but a significance difference was detected between Gulupdo and Anmado; and between Gulupdo and Seogwi-dong only in COI sequences (P < 0.05). Considering the high genetic diversity and gene flow in C. tripartitus populations, one of the major issues regarding conservation seems not to be the recovery of genetic diversity as has been suggested before.

Previous phylogenetic results often showed fluctuating positions of Geometroidea in Macroheterocera, particularly from mitochondrial genome (mitogenome)-based analyses. In this study, we sequenced mitogenomes of four geometrid moths to increase taxon diversity for the inference of phylogenetic positions of Geometroidea in Macroheterocera. The general genomic features found in Macroheterocera also were found in the four geometrid moths. Phylogenetic analyses using 71 representative mitogeneome sequences in Macroheterocera yielded the consensus superfamilial relationships (((((Bombycoidea + Lasiocampoidea) + Geometroidea) + Noctuoidea) + Drepanoidea) + Mimallonoidea), confirming the sister relationship of Geometroidea to (Bombycoidea + Lasiocampoidea) in both Bayesian Inference and Maximum-likelihood method.

The Japanese oak silkmoth, Antheraea yamamai Guérin-Méneville 1861 (Lepidoptera: Saturniidae), is one of the important natural resources possessing industrial value for silk fiber production. In this study, ten microsatellite markers and two mitochondrial DNA (mtDNA) gene sequences (COI and ND4) were used to investigate the genetic variation and geographic structure of A. yamamai populations in South Korea. Two mtDNA gene sequences revealed very low total genetic variation and resultant low geographic variation, validating to use further variable molecular markers. Population-based FIS, FST, RST, and global Mantel test consistently support that A. yamamai populations are overall well interconnected with a relatively high gene flow. Nevertheless, STRUCTURE analysis using microsatellite data and mtDNA sequences coincidently indicate the presence of two genetic pools in many populations.

The tiny dragonfly, Nannophya pygmaea (Odonata: Libellulidae) is one the smallest dragonflies in the world and listed as a second-degree endangered wild animal in Korea. We developed microsatellite markers and applied selected markers to South Korean populations to understand population genetic characteristics, along with two mitochondrial DNA (mtDNA) gene sequences (COI and ND5). Two mtDNA-based population genetic analysis indicates substantially reduced genetic diversity in an island population (Muuido) compared to others. On the other hand, population-based FST and RST consistently support that N. pygmaea populations are overall well interconnected with a relatively high gene flow. These results may collectively indicate that N. pygmaea populations in South Korea may have rather larger population size than we previously acknowledged based on a single-locus mtDNA sequence and field observation.

The red-spotted apollo butterfly, Parnassius bremeri Bremer, 1864 (Lepidoptera: Papilionidae), has been listed as an endangered species in Korea. We developed microsatellite markers by nest-generation sequencing (NGS), selected 12 markers, and applied the markers to available South Korean populations to understand population genetic characteristics. The genotyping of 40 P. bremeri individuals from three localities in South Korea showed that at each locus, the observed number of alleles ranged from 17 to 43, the observed and expected heterozygosities were 0.84722-0.90556 and 0.76045-0.79208, respectively, and FIS was –0.155 to –0.121. STRUCTURE analysis supported the presence of two genetic pools in all three populations, although an immediate reason for this subdivision is not known. The population based FST, RST, and front wing length collectively suggest that at least Samchuk population in Gangwon Province has a significant distance. Further scrutinized analysis is undergoing.

Dendrolimus superans (Lepidoptera: Lasiocampidae) has long been acknowledged to present in South Korea under the circumstance of a substantial change in species names of Dendrolimus. In the present study, we found that specimens of D. superans collected from South Korea were D. sibiricus, rather than D. superans. Comparison of Korean specimens to the D. superans from Japan and the D. sibiricus from Russia in terms of wing morphology, female and male genitalia, a partial COI gene sequence, and a partial internal spacer sequence 2 (ITS2) consistently supported the presence of D. sibiricus in South Korea. Phylogenetic analyses using concatenated sequences of COI and ITS2 from available individuals of D. sibiricus and D. superans both by Bayesian inference and maximum likelihood methods the Korea samples to be D. sibiricus.

Despite of a substantial effort, the phylogeny of Lepidoptera still has unstable taxa and unresolved relationships particularly form Apoditrysia to Ditrysia (e.g. phylogenetic position and familial relationships of Gelechioidea, the monophylies of Tineoidea and Zygaenoidea and so on). We performed phylogenetic analysis using ~13,000-bp length of mitogenome sequences form 50 species (with seven species in two superfamilies form this study) representing 13 superfamilies to improve the lepidopteran phylogeny from Apoditrysia to Ditrysia. A close relationship of Gelechioidea to Obtectomera than to apoditrysian taxa was noteworthy, although the nodal support is very weak (ML, 23%; BI, BPP = 0.94). Monophyly of Zygaenoidea without Epipyropidae (Epipomponia nawai) was also noteworthy in spite of weak nodal support (ML, 64%; BI, 1.0). Scrutinized analysis is currently underway.

We sequenced the complete mitochondrial genome (mitogenome) of Vespa velutina nigrithorax and V. ducalis (Hymenoptera: Vespidae). The genomes were 16,475-bp and 15,779-bp long and contained typical sets of genes. The V. velutina and V. ducalis A+T-rich region was 132-bp long and 166-bp long and was the shortest of all sequenced Vespoidea genomes. Start and stop codons in several Vespa species—including V. velutina and V. ducalis—were diversified, despite these species belonging to the same genus. In comparison with the ancestral mitogenomes, Vespa mitogenomes showed substantial gene rearrangement; however, we detected no gene rearrangement among Vespa species. We conducted phylogenetic reconstruction based on concatenated sequences of 13 PCGs and two rRNAs in available species of Vespoidea—22 species in six subfamilies in two families (Vespidae and Formicidae). The Bayesian inference and maximum likelihood (ML) methods revealed that each family formed strong monophyletic groups.

Currently, only limited number of mitochondrial genomes (mitogenome) is available from Odonata. In order to extend current mitogenome data for comparative biology and phylogeny we sequenced complete mitogenomes of two endangered dragonfly species, Libellula angelina and Nannophya pygmaea (Ododana: Libellulidae). The whole genomes were 15,233 bp in L. angelina and 15,112 bp in N. pygmaea and included a typical set of genes (13 protein-coding genes, two rRNA genes, and 22 tRNA genes) and one major non-coding A+T-rich region. The arrangement of the genomes was identical to typical one found in insects. Phylogenetic reconstruction using concatenated sequences of 13 PCGs and two rRNAs of Odonata (17 species in eight families in three suborders) using both Bayesian Inference (BI) and Maximum Likelihood (ML) methods have shown a strong support for monophyletic Zygoptera (BI, BPP = 1 and ML, 100%). Currently, further scrutinized analysis is under progress.

The number of reported mitochondrial genomes (mitogenomes) from the monotypic Lasiocampoidea has been limited until recently. In this study, we sequenced the complete mitogenome of the lappet moth, Kunugia undans (Lepidoptera: Lasiocampidae), and compared it to those of other lasiocampid species and macroheteroceran superfamilies (59 species in six superfamilies). The 15,570-bp long K. undans genome had the typical set of genes found in animal mitogenomes, with the exception of one additional trnR that are located between trnA and trnN loci. Considering that the two trnR copies are located in tandem with proper secondary structures and identical anticodons, a gene duplication event might be responsible for the presence of the two tRNAs. In summary, the general mitogenome characteristics of Lasiocampoidea did not differ greatly from the remaining macroheteroceran superfamilies, but it did exhibit some unique features.

The spotted-wing drosophila (SWD), Drosophila suzukii (Diptera: Drosophilidae), is an economically damaging pest that feeds on most thin-skinned fruits. In this study, we sequenced portions of the mitochondrial (mt) COI and ND4 genes from a total of 195 individuals collected mainly from Korea. A total of 139 haplotypes were obtained from the concatenated COI and ND4 sequences. A dataset combining GenBank sequences with our own data identified a total of 94 worldwide COI haplotypes with a maximum sequence divergence of 5.433% (32 bp). A rough estimate of genetic diversity in each country showed higher diversity in ancestral distributional ranges, but the invasion over Asian countries seems to have been substantial because haplotype diversity was only 2.35-3.97-fold lower in the USA, Canada, and Italy than that in the populations ancestral ranges.

Currently, phylogenetic relationships of Macroheterocera with the inclusion of Bombycoidea have been reconstructed using molecular data, but generated several hypotheses on familial relationships within Bombycoidea and among-superfamilial relationships within Macroheterocera. Thus, in this study, mitogenome-based superfamilial relationships of Macroheterocera and familial relationships among macroheteroceran superfamily, Bombycoidea, were tested with different algorithms and partition schemes. Additionally, two unrecorded species of Bombycoidea (Saturnia jonasii and Kentrochrysalis streckeri) detected in Korea using morphological and molecular data are presented.

We sequenced complete mitochondrial genomes (mt genomes) of four Apis species, such as A. cerana, A. dorsata, A. laboriosa, and A. mellifera ligustica. The A. m. ligustica used in this study was originated from United States and selected for a high hygienic behavior during several years of breeding program. Considering current phylogenetic hypothesis of Apis the positive relationships between phylogeny and gene rearrangement was not supported. Phylogenetic reconstruction using available Apoidea species has shown the relationships ((Apidae + Colletidae) + Crabronidae). Within Apis, a strong support for three groups was obtained: A. florea and A. andreniformis as a basal group to the other A. cerana and A. mellifera group and A. dorsata and A. laboriosa group. The test to find out suitable single mt gene for the phylogeny at the genus level and below-genus level of Apis provided an identical topology from ND4L and ND6 in the ML analysis and ND3 and NF4L in the BI analysis to that from whole mt genome, with relatively strong support, indicating that these individual genes can potentially be utilized for within-species level phylogeny of Apis.

In order to understand evolutionary characteristics of gene rearrangement in Lepidoptera, we collected all available complete mitogenome (mitogenome) sequences registered in GenBank (274 mitogenomes from 44 families in 23 superfamilies as of August 6, 2015). It turned up six rearrangements that differ from the arrangement of ancestral insects, including that of the gelechioid Mesophleps albilinella that we sequenced in this study. The M. albilinella mitogenome has a unique gene arrangement among the Gelechioidea: ARNESF (the underline signifies an inverted gene) at the ND3 and ND5 junction, as opposed to the ARNSEF that is found in ancestral insects. Most of the rearrangements can be explained by the tandem duplication-random loss model, but inversion, which requires recombination, is also found in two cases, including M. albilinella. Excluding the MIQ rearrangement at the A+T-rich region and ND2 junction, which is found in nearly all Ditrysia, most of the remaining rearrangements found in Lepidoptera appear to be independently derived in that they are automorphic at several taxonomic scales. Current mitogenomic data are limited, particularly for congeneric data. Thus, future research focused on congenerics could clarify evolutionary independency at the generic level also.

The spotted-wing drosophila Drosophila suzukii (Diptera: Drosophilidae) is an Asian species introduced into North America and Europe. It damages a wide variety of thin-skinned fruits. We sequenced the complete mitochondrial genome (mitogenome) of D. suzukii to better understand the mitogenomic characteristics of this species and understand phylogentic relationships of Drosophila. The 16,230-bp complete mitogenome of the species consists of a typical set of genes, including 13 protein-coding genes (PCGs), two rRNA genes, and 22 tRNA genes, and one major non-coding A+T-rich region, with an arrangement typical of insects. Twelve PCGs began with the typical ATN codon, whereas the COI began with TCG, which has been designated as the start codon for other Drosophila species. The 1,525-bp A+T-rich region is the second longest in Drosophila species for which the whole mitogenome has been sequenced, after D. melanogaster. Phylogenetic analysis with the 13 PCGs of the Drosophila species using Bayesian Inference and Maximum likelihood methods both placed D. suzukii at the basal lineage of the previously defined Melanogaster group, with a strong support.

In this study, we tested the effect of a range of insect orders including Trichoptera as outgroups for lepidopteran phylogeny. Phylogenetic analyses performed with four different partitioning schemes using the maximum-likelihood method provided four different topologies (T1-T4) and topological test most supported T1 topology. When the means of first principle component for nucleotide frequency between A/T and G/C of PCGs was considered Trichoptera, Diptera, Coleoptera, and Orthoptera tended to result in T1 topology more frequently in the given ingroup taxa and outgroups tested. This result contradicts to the general view that the sister taxon might be the best outgroup. The T1 topology was largely consistent with a recent large molecular dataset-based lepidopteran phylogeney, presenting the relationships ((((((((((Noctuoidea + Geometroidea) + (Bombycoidea + Lasiocampoidea)) + Drepanoidea) + Mimallonoidea) + Pyraloidea) + Gelechioidea) + Papilionoidea) + Tortricoidea) + (Gracillaroidea + Yponomeutoidea)) + Hepialoidea).

We sequenced the complete mitochondrial (mt) genome of Camponotus atrox (Hymenoptera: Formicidae) that is distributed only in Korea. This genome is 16,540 bp in size, contains typical sets of genes (13 protein-coding genes, 22 tRNAs, and two rRNAs). The C. atrox A+T-rich region is the longest in the sequenced ants as 1,402 bp and is comprised of an identical tandem repeat consisting of six 100-bp copies and one 96-bp copy. A total of 315 bp of intergenic-spacer sequences were spread over 23 regions. An attempt to align spacer sequences in ants turned out that alignment was mostly feasible among congeneric species, with a substantial sequence divergence, indicating the potential of these sequences as congeneric molecular markers. The A/T content in first and second codon positions of PCGs are similar in ants including C. atrox (73.9 vs. 72.3% on average). Estimation of degree of genetic divergence (e.g. non-synonymous substitution rate) with an increased taxon sampling among hymenopteran superfamilies indicated the presence of different rates of divergence between the suborders Symphyta and Apocrita as has previously been reported. The C. atrox mt genome has a unique gene arrangement, trnI-trnM-trnQ at the A+T-rich region and ND2 junction (underline for inverted gene), possibly originated from tandem duplication of trnM-trnI, resulting in trnM-trnI-trnM-trnI-trnQ and loss of first trnM and second trnI, resulting in trnI-trnM-trnQ.

Saturnia (Rinaca) jonasii Butler, 1877 is distributed in Japan, including Tsushima Island and Taiwan, whereas S. boisduvalii Eversmann, 1846 is distributed in northern areas, such as China, Russia, and South Korea. In the present study we found that the specimens from Mt. Hallasan on Jejudo, a southern remote offshore island, were S. jonasii, rather than S. boisduvalii based on morphology, DNA barcode, and nuclear elongation factor 1 alpha (EF-1α) sequences. The major morphological differences between the two species included the shape of wing pattern elements of fore- and hindwings and male and female genitalia. A DNA barcode analysis of the sequences of the Jejudo specimens and S. boisduvalii, along with those of Saturnia species obtained from a public database showed a minimum sequence divergence of 4.26% (28 bp). A phylogenetic analysis also showed clustering of the Jejudo specimens with S. jonasii, separating S. boisduvalii (Bayesian posterior probability = 0.99). The EF-1α-based sequence and phylogenetic analyses of the two species from Jejudo Island and the Korean mainland showed the uniqueness of the Jejudo specimens from S. boisduvalii collected on the Korean mainland, indicating distribution of S. jonasii on Jejudo Island in South Korea, instead of S. boisduvalii.