To date there have been only two species in genus Pogonus and subgenus Europhilus of genus Agonum recognized from Korean insect fauna: Pogonus itoshimaensis Habu, 1954 and Agonum (Europhilus) bellicum Lutshnik, 1934. In the present study one additional species for each genus is newly recognized from Korea: Pogonus (Pogonus) japonicus Putzeys, 1875 and Agonum (Europhilus) gratiosum nipponicum Habu, 1972. A key to adults, redescription, diagnostic photos of adult and male and female genitalia are provided.

In the present study, we report two lepidopteran pests on the cones of Abies koreana E. H. Wilson; Cydia sp. of Tortricidae and Dioryctria abietella Denis et Schiffermüller of Pyralidae. Larvae of these species bored tunnels inside the immature cones of the host plant. Damaged cones are easy to spot by reddish brown excretion from the pest entry holes, and the cones severly damaged by the larvae becomes crooked. Damage rate on the cones reached up tp 71% from the collection site. Regarding the two pests, some basic information such as collection records, morphological characters, and ecology were provided.

Insect pollinators of the endanger orchid Cypripedium japonicum were surveyed and identified during two years, as a part of a conservation project of the orchid at Jukyeup-san and Hwaak-san (Mt.), South Korea. In total 40 individuals of 16 species in 4 families were identified. The dominant family was Halictidae, and Lasioglossum exiliceps Vachal visited the most frequently C. japonicum during the surveys. The average visiting frequency was 2.5 individuals per hour and the highest 4.3, from 12:00 – 13:00 in a day. After 15:00 insects did not visit the flowers at all. However, all of the visiting insects were found to not carry a pollinium or pollens of the orchard on their bodies; pollen carryover by any of the visiting insects did not occur at all. The orchid seems to require certain pollinators in particular body thickness due to its unique pollination mechanism. The orchid has two exit route openings, around 1 cm in diametre, where the entrapped insects can exit and an anther is situated just in front of each opening. It was inferred that a pollen carrier should be around 1 cm in body thickness. Therefore, the candidate species as the proper pollen carriers can be Tetralonia nipponensis Perez, Xylocopa appendiculata circumvolans Smith and Bombus consobrinus Dahlbom among the surveyed visitors.

A partial sequence of the mitochondrial cytochrome oxidase subunit I (COI) gene is widely used as a molecular marker for species identification in animals, also termed a DNA barcode. However, the presence of more than one sequence type in a single individual, also known as heteroplasmy, is one of the shortcomings of barcode identification. In this study, we examined the extent and divergence of COI heteroplasmy, including nuclear-encoded mitochondrial pseudogenes (NUMTs), at the genomic-DNA level from 13 insect species, including four individuals of orthopteran Anapodisma miramae. Furthermore, a long fragment of mitochondrial DNA (~13.5 kb) and cDNA from A. miramae were used as a template for COI PCR to compare the patterns of heteroplasmy between DNA sources and to investigate a possible way to avoid ambiguity in DNA barcoding. When multiple numbers of clones originated from genomic DNA were sequenced, heteroplasmy was prevalent in all species (3~16 heteroplasmic copies), with a varying degree of maximum sequence divergence (<1% in 7 species, <4% in 3 species, <6% in 2 species and 2.15-8.03% in four A. miramae individuals). In five species, NUMTs also were observed when genomic DNA was used as a template. Long fragment DNA also is a source of heteroplasmic amplification, but the divergent haplotypes and NUMTs obtained in the genomic DNA-based PCR were not detected in A. miramae. On the other hand, cDNA was heteroplasmy-free, without NUMTs when multiple numbers of clones were sequenced. Consistently, one dominant haplotype was always obtained from the genomic DNA-origin clones in all species and also from the long fragment- and cDNA-origin clones of A. miramae. Furthermore, the dominant haplotype was identical in sequence, regardless of the DNA source. Thus, one possible solution to avoid the barcoding problem in relationship to heteroplasmy could be the acquisition of multiple numbers of barcoding sequences to determine a dominant haplotype that can be assigned as barcoding sequence for a given species.

The sister relationship between Trichoptera and Lepidoptera has often been supported in a diverse study, but mitochondrial genomes (mitogenomes) based lepidopteran phylogenetic studies have never utilized Trichoptera as outgroup mainly due to unavailability. Therefore, the effect of alternatives that were previously used (e.g., Diptera, Hymenoptera, and Orthoptera) or Trichoptera as outgroups on the lepidopteran phylogeny remained unknown. In this study, we sequenced three complete mitogenomes of Trichoptera belonging to two suborders and characterized the genomic features of Trichoptera and tested the outgroup effect for lepidopteran phylogeny. The 15,208 ~ 15,285-bp long caddisfly mitogenomes harbor gene content typical of the animal mitogenomes. The orientation and gene order of the three species belonging to the suborder Integripalpia was identical to that of the most common type that has been hypothesized as ancestral for insects, but Cheumatopsyche brevilineata belonging to another suborder Annulipalpia has rearranged QIM, all encoded in forward direction between the A+T-rich region and ND2, instead of the ancestral IQM, with Q inverted. Further, the annulipalpian species had a typical start codon ATG, instead of CGA that are commonly found in other trichopteran species and majority of Lepidoptera. Phylogenetic analysis with different outgroups (Diptera, Hymenoptera, and Orthoptera, Coleoptera, and Trichoptera) and 115 lepidopteran mitogenomes has shown insensitivity either with Trichoptera, Diptera, or Coleoptera, but artificial grouping and lowered nodal support were found with Hymenoptera. The Trichoptera-based consensus topology were: (((((((Bombycoidea + Noctuoidea) + Pyraloidea) + Papilionoidea) + Cossoidea) + Tortricoidea) + Yponomeutoidea) + Hepialoidea).

The mulberry white caterpillar, Rondotia menciana, belongs to the lepidopteran family Bombycidae, in which the domestic silkworm, Bombyx mori is included. In this study, we describe the complete mitochondrial genome (mitogenome) sequences of the species in terms of general genomic features and characteristic features found in the A+T-rich region. The 15,364-bp long genome consisted of 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, with the typical arrangement found in Lepidoptera. Twelve of the 13 PCGs start with typical ATN codons, except for the COI, which begins with CGA. Twelve of the 13 PCGs have complete stop codon, except for COII, which ends up with a single T. The 360-bp long A+T-rich region harbored the conserved sequence blocks that are typically found in lepidopteran insects. Additionally, the A+T-rich region of R. menciana contained one tRNAMet-like structure, which has a proper anticodon and secondary structure.

We newly sequenced mitogenomes of five skippers belonging to Lepidoptera to obtain further insight into characteristics of butterfly mitogenomes and performed phylogenetic reconstruction using all available gene sequences (PCGs, rRNAs, and tRNAs) from 85 species in 19 families in eight superfamilies. The general genomic features found in the butterflies also were found in the five skippers: a high A/T composition (79.3% - 80.9%), dominant usage of TAA stop codon, similar skewness pattern in various levels, consistently long intergenic spacer sequence between tRNAGln - ND2 (64-87 bp), the ATACTAA motif betweent RNASer(UCN) and ND1, and characteristic features of the A+T-rich region (the motif ATAGA, varying length of poly-T stretch, and poly-A stretch). The start codon for COI was CGA in four skippers as typical, but Lobocla bifasciatus evidently possessed canonical ATG as start codon. Phylogenetic analyses mainly yielded the consensus superfamilial relationships ((((((Bombycoidea + Noctuoidea + Geometroidea) + Pyraloidea) + Papilionoidea) + Tortricoidea) + Yponomeutoidea) + Hepialoidea) with a high support for most nodes, confirming the validity of Macroheterocera and its sister relationship to Pyraloidea. Within Rhopalocera the familial relationships (Papilionidae + (Hesperiidae + (Pieridae + ((Lycaenidae + Riodinidae) + Nymphalidae))) were strongly supported, confirming invalidity of the superfamily Hesperioidea. On the other hand, superfamilial relationships among Noctuoidea, Geometroidea, and Bombycoidea and the familial relationships among Saturniidae, Sphingidae, and Bombycidae were dubious, requiring further representative taxon sampling.

We report the complete mitochondrial genome (mitogenome) of Apodemia mormo, which belongs to the lepidopteran family Riodinidae. The 15,262-bp long complete genome is comprised of 13 protein-coding genes, two rRNA genes, 22 tRNA genes, and one major non-coding A+T-rich region, with the arrangement typically found in majority of Lepidoptera. The genes of A. mormo are interleaved with a total of 168 bp, which are spread over 16 regions and overlap in a total of 58 bp at eight locations. All tRNAs of the A. mormo mitogenome formed typical cloverleaf structure, except for tRNASer(AGN), which formed the truncated dihydrouridine arm. COI gene started with CGA, instead of canonical ATN as seen in other Lepidoptera. The 349-bp long A+T-rich region harbored the conserved sequence blocks, such as ATAGA motif, poly-T stretch, the conserved ATTTA sequence, and microsatellite A/T repeat that are typically found in Lepidoptera, but absent for tRNA-like pseudogene.

The multi-tactile stimulation system was developed to analyze cognitive function characteristics. The developed system consists of three units; control unit, drive unit and actuator that simultaneously and separately supports the vibration, heat, and pressure stimulation. From the results, vibration stimuli have a wide frequency range of 0-300 Hz comprising 20 levels, and stimulation intensity comprising 40 levels. The control temperature and stimulation temperature used for heat stimulation comprise four and eight levels, respectively. Pressure stimuli in the range of 0-600 mmHg can be stimulated comprising 12 levels. Therefore, the developed stimulator can be used for integrative tactile perception and cognition studies on various vibration, heat and pressure sensations.

In this paper, we developed a power amplifier system for driving a solenoid coil. The developed system consists of three units; control unit, drive unit and coil. The control unit generates a drive signal of sinusoidal wave type, monitors an operating status of the drive unit and vibration characteristics of the coil. The drive unit drives a solenoid coil in response to command from the control unit. The coil was fabricated with the size of diameter in 1.5 cm and length in 1.7 cm using electric copper wire. 4 types of drive unit were tested with thermal and vibration characteristics of the coil using temperature sensor and 3-axis accelerometer, respectively. From the result, the developed system has many advantages with respect to simple control algorithm, small size, and operates stably up to 68 watts driving condition.

We newly sequenced mitochondrial genomes of Spodoptera litura and Cnaphalocrocis medinalis (Lepidoptera) to obtain further insight into mitochondrial genome evolution and investigated the influence of optimal strategies on phylogenetic reconstruction of Lepidoptera. Estimation of p-distances of each mitochondrial gene for available taxonomic levels has shown the highest value in ND6, whereas the lowest values in COI and COII at the nucleotide level, suggesting different utility of each gene for different hierarchical group when individual genes are utilized for phylogenetic analysis. Phylogenetic analyses mainly yielded the relationships (((((Bombycoidea + Geometroidea) + Noctuoidea) + Pyraloidea) + Papilionoidea) + Tortricoidea), evidencing the polyphyly of Macrolepidoptera. The tests of optimality strategies, such as exclusion of third codon positions, inclusion of rRNA and tRNA genes, data partitioning, RY recoding approach, and recoding nucleotides into amino acids suggested that the majority of the strategies did not substantially alter phylogenetic topologies or nodal supports, except for some familial relationship only in the amino acid dataset.

Glyphodes quadrimaculalis (Lepidoptera: Crambidae) feed on a root tuber of Cynanchum wilfordii (Asclepiadaceae) that is one of the most famous traditional medicines in Korea. The genus Glyphodes includes ~130 species distributed worldwide, so the complete mitochondrial genome (mitogenome) would be helpful for bio-identification, biogeographic studies, and multigene-based phylogeny. The 15,255-bp long G. quadrimaculalis genome comprises 37 typical genes and one large non-coding region, with the typical arrangement found in Lepidoptera. Of the 13 protein coding genes (PCGs), 12 begin with typical start codons found in insect mitochondrial PCGs, but the COI gene starts with atypical CGA. One of the noteworthy features of the genome includes the presence of a 51-bp long non-coding space sequence located between tRNAGln and ND2 that reveals high sequence homology (71.4%) to the neighboring ND2 gene, indicating the origin of the region by partial duplication of the ND2 gene.

We present the nearly complete mitogenome sequences of the garden chafer, Polyphylla laticollis manchurica, which is listed as an endangered species in Korea. The P. l. manchurica mitogenome, which includes unfinished whole A+T-rich region and a partial srRNA was 14,473-bp long, possessing typical sets of genes (13 PCGs, 22 tRNA genes, and 2 rRNA genes). Gene arrangement of the P. l. manchurica mitogenome was identical to the common one found in the majority of insects. The 5 bp-long motif sequence (TAGTA) that has been suggested to be the possible binding site for the transcription termination peptide for the major-strand was also found in the P. l. manchurica mitogenome between tRNASer(UCN) and ND1. As has been previously determined, the high A/T content was unanimously observed in P. l. manchurica in terms of A/T bias in the third codon position (73.5%) compared with the first (66.4%) and second codon position (66.2%), and a high frequency of A/T-containing codons (a total of 28.22% for TTA, ATT, TTT, and ATA). The PCGs encoded in major-strands are slightly T-skewed, whereas those of the minor-strand are A-skewed, indicating strand asymmetry in nucleotide composition in the Coleoptera including P. l. manchurica.

We sequenced 17,329 bp of mitochondrial genome (mitogenome) of the black dwarf honey bee, Apis andreniformis (Hymenoptera: Apidae), that lacks ~200 bp of the A+T-rich region for the completion of the genomic sequence. The gene arrangement of A. andreniformis mitogenome is identical to that of A. cerana. However, the genome contains 5 additional tRNALeu(CUN) located 4 copies between tRNAMet and tRNAGln, and 1 copy between tRNAGln and tRNAAla, along with the typical sets of genes (13 protein-coding genes, 22 tRNAs, and 2 rRNAs) including regular tRNALeu(CUN) and the A+T-rich region (at least 923 bp). Only 1 copy of tRNALeu(CUN) differed by 1 bp from other 4 copies of tRNALeu(CUN). Each additional tRNALeu(CUN) is followed by nearly identical 68-bp long repeat sequence (95.6% identity). All 13 protein coding genes have typical start codons found in insect mitochondrial PCGs (2 ATA, 9 ATT, and 2 ATG).

Hipparchia autonoe belongs to the family Nymphalidae (Lepidoptera) and is designated as an endangered insect and national monument in Korea. It only inhabits a very restricted area on Mt. Halla but is widely distributed in several Asian countries including Mongolia. A previous study conducted to understand the genetic relationship between Mt. Halla and Mongolian H. autonoe for conservation purposes suffered from a limited number of samples. Therefore, we sequenced the DNA barcode region of an additional 36 H. autonoe individuals, combined them with previous data from 19 individuals, and performed phylogenetic and population genetic analyses. Furthermore, the internal transcribed spacer 2 (ITS2) region was also sequenced from the 36 samples as a nuclear DNA marker. The existence of independent haplotypes, sequence types, and significant FST estimates (p < 0.05) between Mt. Halla and Mongolian populations indicated hampered gene flow between the populations. Nevertheless, an absence of a reciprocal monophyletic group in Mt. Halla and Mongolian populations by cytochrome oxidase subunit I gene- and ITS2-based phylogeny suggests that the genetic isolation of the Mt. Halla population from the Mongolian populations seemed not large enough to consider them independent genetic entities.

The species status of Oeneis urda (Eversmann) and O. mongolica (Oberthür) has been argued based on morphological characters. Reexamination of their major morphological characters has shown a slight differentiation in the two species. Sequences of three mitochondrial genes (COI, ND6, and ND1) and one nuclear region (internal transcribed spacer 2, ITS2) from two O. urda populations (Yangyang and Mt. Hanla) and one O. mongolica population (Uljin) were performed for phylogenetic and population genetic inferences. Sharing of identical sequences in the ND6 gene and ITS2, minimal sequence divergence in the COI and ND1 genes, and phylogenetically undividable sequence types in all mitochondrial genes and ITS2 suggest genetic continuity between the two species. Nevertheless, significant FST estimates (p < 0.05) were found for the COI gene in comparisons between Yangyang (O. urda) and Uljin (O. mongolica), between Yangyang (O. urda) and Mt. Hanla (O. urda), and between Uljin (O. mongolica) and Mt. Hanla (O. urda) populations. These FST estimates, along with other gene-based analyses collectively suggest isolation of the two species at some point in the past, but incomplete separation between the two species on the mainland (Yangyang and Uljin) and biogeographically forced isolation of the O. urda population on Mt. Hanla collectively appear to complicate species status of these two species that were once further clearly separated.

Lepidoptera is one of the largest insect orders, but the phylogenetic relationships within this order, have yet to be completely described. One of the unresolved relationships includes the monophyly of Papilionoidea in relationship with the monotypic superfamily Hesperioidea. We newly sequenced five hesperid mitochondrial genomes (mitogenomes), representing four subfamilies: Pyrginae (Daimio tethys and Lobocla bifasciatus), Coeliadinae (Choaspes benjaminii), and Hesperiinae (Potanthus flavus), and Heteropterinae (Carterocephalus silvicola). Along with these newly sequenced hesperid genomes phylogenetic analysis was conducted with all available lepidopteran mitogenomes including three reported species of Hesperiidae that consisted of ~70 species in ten lepidopteran superfamilies. The test for the effect of optimization schemes, such as exclusion and inclusion of third codon position of 13 PCGs, other genes (22 tRNAs and two rRNAs), and with and without partitions also was performed. Majority of datasets consistently placed the monophyletic Hesperiidae the sister to ((Pieridae + Lycaenidae) + Nymphalidae), placing another true butterfly family Papilionidae as the basal lineage of this group, presenting the relationships (Papilionidae + (Hesperiidae + ((Pieridae + Lycaenidae) + Nymphalidae))). Consistent to previous result, Pyraloidea was placed as the sister to ((Bombycoidea + Geometroidea) + Noctuoidea), placing the Macrolepidoptera as non-monophyletic group.

The larch hawk moth, Sphinx morio, belongs to the lepidopteran family Sphingidae that has long been studied as a family of model insects in a diverse field. In this study, we describe the complete mitochondrial genome (mitogenome) sequences of the species in terms of general genomic features and characteristic short repetitive sequences found in the A+T-rich region. The 15,299-bp long genome consisted of 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, with the typical arrangement found in Lepidoptera. The 316-bp long A+T-rich region located between srRNA and tRNAMet harbored the conserved sequence blocks that are typically found in lepidopteran insects. Additionally, the A+T-rich region of S. morio contained three characteristic repeat sequences that are rarely found in Lepidoptera: two identical 12-bp repeat, three identical 5-bp long tandem repeat, and six nearly identical 5~6 bp long repeat sequences.

The bumblebee, Bombus ignitus (Hymenoptera: Apidae), is a valuable natural resource that is widely utilized for greenhouse pollination in South Korea. Understanding the magnitude of genetic diversity and geographic relationships is of fundamental importance for long term preservation and utilization. As a first step, we sequenced a partial COI gene of mitochondrial DNA (mtDNA) corresponding to the “DNA barcode” region and the complete internal transcribed spacer 2 (ITS2) of nuclear ribosomal DNA from 88 individuals collected in nine South Korean localities. The complete ITS2 sequences were longest among known insects, ranging in size from 2,034 bp ~ 2,052 bp, harboring two duplicated 112-bp long repeats. The 658-bp long mtDNA sequences provided only six haplotypes with a maximum sequence divergence of 0.61% (4 bp), whereas the ITS sequences provided 84 sequence types with a maximum sequence divergence of 1.02% (21 sites). The combination of the current COI data with those of published data suggest that the B. ignitus in South Korea and China are genetically a large group, but those in Japan can be roughly separated into another group. Overall, a very high per generation migration ratio, a very low level of genetic fixation, and no discernable hierarchical population were found to exist among the South Korean populations of B. ignitus, which suggests panmixia. This finding is consistent with our understanding of the dispersal capability of the species.