The pear psylla, Cacopsylla pyricola Foerster (Homoptera: Psyllidae), is a serious insect pest of commercial pear crops. The species, which resides on pear trees throughout its life cycle, is rapidly spreading in some regions of the world. Given the life cycle, it is unclear how such a rapid spread has been facilitated. Presently, the population genetic structure of the species including genetic diversity and gene flow was studied to understand the nature of dispersal and field ecology of the species. Pear psylla was collected from several pear orchards in Korea. The 658-bp region of mitochondrial COI gene and the 716-bp long complete internal transcribed spacer 2 (ITS2) of the nuclear ribosomal DNA were sequenced. Unlikely other previously studied insect pests, the COI-based genetic diversity of the pear psylla was extremely low (maximum sequence divergence of 0.15%). This finding allowed us to conclude that the species may have been introduced in Korea relatively recently, possibly with the phenomenon of genetic bottlenecks. ITS2 sequence-based analyses of phylogeny, population differentiation, gene flow, and hierarchical population structure all concordantly suggested that the pear psylla populations in Korea are neither genetically isolated nor hampered for gene flow. These genetic data are concordant with the dispersal of an overwintering winterform morph outside the non-pear habitat in the fall and the possibility of subsequently longer distant dispersal.

The phylogenetic relationships among the Nymphalidae (Lepidoptera: Papilionoidea) have been controversial. The present study sequenced approximately 1,099 bp from cytochrome oxidase subunit I (COI), 1,336 ~ 1,551 bp from 16S ribosomal RNA (16S rRNA), and 1,066 bp from elongation factor-1 alpha (EF-1α) in 80 species belonging to seven subfamilies (Linmenitidinae, Heliconiinae, Nymphalinae, Apaturinae, Libytheinae, Satyrinae, and Danainae) of Nymphalidae, along with those of six lycaenid species as outgroups. The average base compositions for the three genes (COI, 16S rRNA, and EF-1α) are as follows: A (30.6%, 38.8%, and 25.8%), G (14.7, 5.2%, and 23.6%), T (39.8%, 45.2%, and 23.4%), and C (14.9%, 10.8%, and 27.3%). This result shows the A/T bias in the mitochondrial genes, but not for the nuclear EF-1α. Between the two mitochondrial genes, the 16S rRNA gene evidenced a significantly higher A/T content than was detected in the COI gene. These sequences were subjected to phylogenetic reconstruction via Bayesian Inference (BI) and Maximum Likelihood (ML) algorithms. Both analyses concordantly supported the subfamilial relationships of (((((Linmenitidinae + Heliconiinae) + (Nymphalinae + Apaturinae)) + Libytheinae) + Satyrinae) + Danainae), along with highly supported monophyletics of tribes within subfamilies. This result is largely consistent with a previous study performed with a large sequence information and morphological characters, except for the position of Libytheinae, which was suggested to be the basal lineage of Nymphalidae.

Two complete mitochondrial genomes of the tobacco cutworm, Spodoptera litura (Lepidoptera: Noctuoidea) and the rice leaf roller, Cnaphalocrocis medinalis (Lepidoptera: Pyralidae), were sequenced. Each 15,388 bp and 15,368 bp-long genome contained both the lepidopteran specific gene arrangement that differ from the most common arrangement of insects by the movement of tRNAMet to a position 5’-upstream of tRNAIle. Neither of the species have typical COI start codon. Instead, the CGA (arginine) sequence that is commonly present in other lepidopterans was also found both in S. litura and C. medinalis. The evolutionary rates among 13 protein-coding genes (PCGs) in Lepidoptera showed ATP8 the highest, whereas COI the lowest. The high A+T-content, which is characteristic of mitochondrial genome was well reflected in the two lepidopteran mitochondrial genomes: higher frequency of A/T-rich codons, severe A/T bias in 3rd codon position, and extremely high A/T content in the A+T-rich region. Because insect mitochondrial genomes harbor biased nucleotide and resultantly biased amino acid sequences, phylogenetic inference is often misled by them. Although each recoded and unrecoded datasets for nucleotide sequences and amino acid sequences of PCGs provided overall identical topology, regardless of recoded scheme, each nucleotide and amino acid dataset provided difference in the status of Macrolepidoptera, providing a monophyletic group by amino acid dataset, whereas non-monophyletic group by nucleotide dataset.

Attacin is a well-studied glycine-rich antibacterial protein in insect immune response, which has limitary antibacterial effect to some Gram-negative bacteria. A cDNA encoding the attacin gene was screened and isolated from the immunized larvae of the swallowtail butterfly, Papilio xuthus. The complete P. xuthus attacin cDNA is 949 nucleotides encoding a 250 amino acid precursor that contains a putative 18-residue signal peptide, a common 42-residue propeptide sequence and a presumed 190-residue mature protein with a theoretical mass of 19904.01 and a pI of 9.13. The putative mature protein of P. xuthus attacin showed 48%~52% and 24%~30% identity in amino acid sequences with that of lepidopteran and dipteran insects, respectively. The attacin transcript was induced at significant level after injection with bacterial lipopolysaccharide (LPS). Recombinant attacin was highly expressed in E. coli BL21 (DE3) cells by fusing with an N-terminal S-tag/thrombin cleavage site configuration protein to avoid the cell death during induction. The expressed fusion protein was purified by Ni-NTA immobilized metal affinity chromatography (IMAC). After desalting and cleavage with thrombin, the recombinant attacin was released and showed considerably antibacterial activity against Gram-negative bacteria, E. coli ML 35. Our results proved that this protein family with a potent antibacterial activity may play a role in the immune response of butterflies.

The perilla leaf pyralid moth, Pyrausta panopealis Walker (Lepidoptera: Pyralidae), is a serious pest damaging to leaf perilla. In order to establish the life parameters of P. panopealis for eventual purpose of control, the developmental span of each stage were investigated under five temperature regimes (20℃ ~ 3 0℃). In addition, the larvicial efficacy of several on-the-market environment-friendly agricultural materials (EFAMs) was tested. The width of head capsule at each larval stage measured to be the mean of 0.21, 0.32, 0.47, 0.64, and 0.98 mm, respectively. The larval period of P. panopealis was longest at 20ºC as 27.0 days and shortened as temperature goes up to 30ºC as 11.3 days. Survivorship of the larval P. panopealis was the highest at 30ºC as 80%, whereas that of other temperatures ranged from 40% (20ºC) to 62.5% (27.5ºC), indicating that the P. panopealis appears to favor higher temperature. In addition to larval period, the duration of egg, prepupa, and pupa stages also shortened as temperature goes up, whereas the duration of adult stage increased as temperature goes up: from 4.1 days at 20 ºC to 6.1 days at 30ºC. After the perilla leaf pyralid moths were successfully stabilized in indoor environment the larvicidal efficacy of the ten EFAMs that were previously selected from the result of other moth species was tested aimed at 4th instar larvae for 48 hrs. Seven of the ten tested showed more than 90% of mortality within 12 hrs and reached nearly up to 100% within 24 hrs, but the remaining three showed less than ~70%.

The phylogenetic relationships among true butterfly families (superfamily Papilionoidea) have been a matter of substantial controversy, and that debate has led to several competing hypotheses. Two of the most compelling of those hypotheses involve the relationships of (Nymphalidae + Lycaenidae) + (Pieridae + Papilionidae) and (((Nymphalidae + Lycaenidae) + Pieridae) + Papilionidae). In this study, approximately 3,500 nucleotide sequences from cytochrome oxidase subunit I (COI), 16S ribosomal RNA (16S rRNA), and elongation factor-1 alpha (EF-1α) were sequenced from 83 species belonging to four true butterfly families, along with those of eight outgroup species belonging to the skipper family (superfamily Hesperioidea). These sequences were subjected to phylogenetic reconstruction via Bayesian Inference (BI), Maximum Likelihood (ML), and Maximum Parsimony (MP) algorithms. All phylogenetic analyses among the four true butterfly families strongly indicated a sister relationship between the Nymphalidae and Lycaenidae on one hand, and relatively strongly indicated a sister relationship between the Pieridae and Papilionidae on another hand, thus supporting the hypothesis: (Nymphalidae + Lycaenidae) + (Pieridae + Papilionidae).

The bumblebee, Bombus ignitus (Hymenoptera: Apidae), is a valuable natural resource that is one of the most notably utilized for greenhouse pollination in Korea. In order to understand the nature of genetic relationships, gene flow, and population structure of the species we sequenced a partial COI gene of mitochondrial DNA (mtDNA) corresponding to “animal barcode” region and the complete internal transcribed spacer 2 (ITS2) of the nuclear ribosomal DNA (nrDNA) collected from Korean localities. Although the 658-bp long mtDNA sequence provided only six haplotypes with the maximum sequence divergence of 0.61% (4 bp), the ITS sequences provided 84 sequence types with the maximum sequence divergence of 1.02% (21 sites), confirming better applicability of the ITS sequences to the study of intraspecific variation. The complete ITS2 sequences of B. ignitus were shown to be longest among known insects, ranging in size from 2,034 bp ~ 2,052 bp, harboring two duplicated repeats. Overall, a very high per generation migration ratio, a very low level of genetic fixation, and no discernable hierarchical population/ population group were noted to exist among populations of B. ignitus on the basis of both molecules, thus suggesting that the B. ignitus populations on the Korean peninsula are panmictic, which is consistent with our understanding of the dispersal capability of the species

In an effort to gain a better understanding of the nature of the population genetic structure of the pest insect, Spodoptera litura (Lepidoptera: Noctuidae), tobacco cutworms were collected from six Korean and five Chinese localities and their mitochondrial A+T-rich region and nuclear internal transcribed spacer 2 (ITS2) regions were cloned and sequenced. A total of 106 A+T-rich region haplotypes and 92 ITS2 sequence types were obtained from 158 individuals. Compared to the preliminary mitochondrial COI gene sequence data that provided all identical sequence types in all samples, the A+T-rich region and nuclear ITS2 are highly variable. Overall, a low level of genetic fixation in the A+T-rich region (FST = 0 ~ 0.02965) and nuclear ITS2 (FST = 0 ~ 0.34491), and no discernable isolated population was noted to exist among most S. litura populations. The presence of a majority of within-population variations, rather than variations among populations or between China and Korea, indicates that the S. litura populations are composed of heterogeneous individuals. The fixation index at hierarchical level of among regions shows no significance, suggesting that the S. litura populations in both Korea and China are profoundly interrelated with one another. This finding is in accord with the current knowledge that S. litura has sufficient flight capacity for dispersal.

Two complete mitochondrial genomes (mitogenomes) of the endangered, lycaenid butterflies, Spindasis takanonis and Protantigius superans (Lepidoptera: Lycaenidae), were sequenced. Each 15,349 bp and 15,248 bp-long genome contained both the lepidopteran specific gene arrangement that differ from the most common arrangement of insects by the movement of tRNAMet to a position 5’-upstream of tRNAIle. Neither of the species have typical COI start codon. Instead, the CGA (arginine) sequence that is commonly present in all other lepidopterans was also found in both lycaenids. The possible binding site for the transcription termination peptide, TACTA sequence, also was well retained in both species. The high A+T-content, which is a characteristic of insect mitogenomes was well reflected in the genomes in the form of higher frequency of codons with A/T nucleotides, severe A/T bias in 3rd codon position, and extremely high A/T content in the A+T-rich region. The 19 bp-long poly-T stretch and the downstream conserved motif ATAG, which were suggested previously to function as a structural signal for minor-strand mtDNA replication, was also well conserved in the A+T-rich region of both lycaenids. Phylogenetic analysis among lepidopteran superfamilies supported the relationships of either (((((Bombycoidea + Geometroidea) + Noctuoidea) + Papilionoidea) + Pyraloidea) + Tortricoidea) by concatenated amino acid sequence or (((((Bombycoidea + Geometroidea) + Noctuoidea) + Pyraloidea) + Papilionoidea) + Tortricoidea) by concatenated nucleotide sequences of 1st and 2nd codon positions of 13 protein-coding genes, two rRNA genes, and 22 tRNA genes, revealing fluctuating positions of Papilionoidea and Pyraloidea between the two data sets.

Diamondback moth (Plutella xylostella L.) belonging to genus Lepidoptera is a notorious pest of cruciferous crops worldwide. We evaluated the bioinsecticidal activity of the liquid cultures (LB and NB) of a bacterial strain, Serratia sp. EML-SE1, isolated from a diseased diamondback moth. The pathogenicity of a bacterial strain to diamondback moth was confirmed by the following procedures: treatment of liquid culture on cabbage leaves, ingestion of inoculated cabbage and mortality response. For the test, twenty 3rd instar larvae of diamondback moth were placed on the Chinese cabbage leaf in a round plastic cage (Ø 10 × 6 cm) and sprayed with the liquid cultures. After 72 hours, insecticidal activity of LB and NB cultures of Serratia sp. against P. xylostella larvae showed 91.7% and 88.3%, respectively. In addition, the bioinsecticidal activity on potted cabbage with 14 leaves in a growth cage (165 × 83 × 124 cm) also was similar to that of plastic cage experiment. Summarized, the Serratia sp. EML-SE1 may be a potent candidate as a bioinsecticidal agent to control diamondbac kmoth.

In organic agriculture, various cover-crops have been used to control weeds. In this study, we have investigated the effect of Vicia tetrasperma (L.) Schred. (Eolchigiwandu) which is native to on major insect pests of pepper. Redpepper seedlings at 8 leaves stage were transplanted in 20th May 2009 into experimental field located in the farm of the department of agricultural biology, Suwon, Korea. A cover-crop cultivation plot was compared with a control plot mulched by black plastic-film. Density of aphids and damaged fruits by oriental tobacco budworm, Helicoverpa assulta, were surveyed ten times from 21 days after transplanting (DAT) to 82 DAT. Fifty and forty pepper plants were sampled to count the density of aphids and damaged fruits by oriental tobacco budworm per a plant, respectively. In current study, three aphid species namely, cotton aphid, Aphis gossypii, green peach aphid, Myzus persicae, potato aphid, Macrosiphum euphorbiae were collected. Overally, the density of aphids on pepper in the control plot was higher than the cover-crop plot. Especially, aphid density increased up to 67.44±26.8 in early stage of control plot, whereas aphid was not found in cover-crop plot. The rate of fruit damage by oriental tobacco budworm was significantly higher in the cover-crop plot than the control plot in early stage of pepper, however damaged fruit rate of pepper in the control plot was significantly higher since the middle of July.

The devastating insect pests of the tomato (Lycopersicon esculentum) were surveyed in environmentally friendly (no-pesticide and organic) greenhouses located in the provinces of Gyeonggi, Gyeongsang, Gangwon, Chungcheong and Jeju. Whitefly, leaf miner, aphid and moth were recorded as the major pests in these surveys. Among them, whitefly was the most serious pest of greenhouse cultivated tomato and major species was Trialeurodes vaporariorum. The population densities of T. vaporariorum was 0~36 nymph per leaf. Population of moths Spodoptera litura were the highest in summer season. Population of the whitefly was considered to be influenced by the previous cultural methods. Farmers in organic cultivation, used insect natural enemy, commercial EFAM (Environment Friendly Agricultural Material), yellow sticky trap or home made pesticides. Commercial microbial pesticides were applied to evaluate their insecticidal activities against whitefly. The results revealed that microbial pesticides exhibited 65.3 to 100% whitefly nymph mortality. Further, pesticidal activities of Cooking Oil and Yolk (COY), plant extracts and neem oil were evaluated against other tomato pests as well.

In order to understand the nature of population genetic structure of the notorious pest insect, Spodoptera litura (Lepidoptera: Noctuidae), a total of 159 individuals of the tobacco cutworm collected from six Korean and five Chinese localities were cloned and sequenced their mitochondrial A+T-rich region. A total of 90 haplotypes ranged 324 bp ~ 372 bp in length and 0.30% ~ 4.85% in sequence divergence. Compared to the preliminary mitochondrial COI gene sequence data that provided nearly identical sequences in all samples, the A+T-rich region provided substantially high sequence variation, validating the applicability of this molecule to the study of intraspecific variation. Overall, a very high per generation migration ratio (Nm = 5.91 ~ infinite), a very low level of genetic fixation (FST = 0 ~ 0.077796), and no discernable isolated population were noted to exist among populations of S. litura, although some significant genetic differentiations were found between several pairs of populations. These results collectively suggest that the S. litura populations in both Korea and China are very well interrelated to each other. The structural analysis revealed that the S. litura A+T-rich region harbored two stretches of the [TA(A)]n sequence, the poly-A stretch, and a stem-and-loop structure that are well found in other lepidopteran A+T-rich region. Furthermore, the region harbors the 13 ~ 19-bp long T stretch and 5-bp long motif that have been suggested to function as a possible recognition site for the initiation of replication of the minor strand of mtDNA also were found.

The bumblebee, Bombus ignitus (Hymenoptera: Apidae), is a valuable natural resource that is one of the most notably utilized for greenhouse pollination in Korea. In order to understand the nature of genetic relationships, gene flow, and population structure of the species we sequenced a partial COI gene of mitochondrial DNA (mtDNA) corresponding to “animal barcode” region and the complete internal transcribed spacer 2 (ITS2) of the nuclear ribosomal DNA (nrDNA) with the individuals collected from Korean localities. Although the 658-bp long mtDNA sequence provided only six haplotypes with the maximum sequence divergence of 0.456% (3 bp, from 91 individuals), but the ITS sequences provided 33 sequence types with the maximum sequence divergence of 0.78% (16 bp, from 35 individuals), confirming better applicability of the ITS sequences to the study of intraspecific variation. The ITS2 sequences of B. ignitus were shown to be the longest among known insects, ranging in size from 2,034 bp ~ 2,045 bp and harbored two duplicated repeats, indicating unusual structure of B. ignitus ITS2 sequences compared with other insect ITS sequences. Overall, a very high per generation migration ratio, a very low level of genetic fixation, and no discernable hierarchical population/population group were noted to exist among populations of B. ignitus on the basis of both molecules, thus suggesting that the B. ignitus populations on the Korean peninsula are panmictic, which is consistent with our understanding of the dispersal capability.

Up to now only 17 coleopteran mitogenomes have been fully or near fully sequenced, but this chart does not include any complete mitogenome sequence of the coleopteran infraorder Scarabaeiformia, in which the monotypic Scarabaeoidea is included. In this study, we firstly present three scarabaeoid mitogenomes, two of which are completed and one is near completed. The complete nucleotide sequences of the white-spotted flower chafer, Protaetia brevitarsis (Scarabaeiformia: Scarabaeidae), was determined to be 20,319 bp, the two-spotted stag beetle, Metopodontus blanchardi (Scarabaeiformia: Lucanidae), was 21,678 bp, and the incomplete garden chafer, Polyphylla laticollis (Scarabaeiformia: Scarabaeidae), which lacks sequence information from whole A+T-rich region and a partial srRNA gene was 14,473 bp. Thus, the two complete mitogenome sequences are longest among the completely sequenced insect mitogenomes, followed by the 19,517 bp-long dipteran Drosophila melanogaster. Phylogenetic analysis using the concatenated 13 amino acid sequences of PCGs of the 13 coleopteran insects including three newly sequenced scarabaeoid successfully recovered a monophyly of suborder Polyphaga, a monophyly of infraorder Cucujiformia, a monophyly of infraorder Elateriformia, and also a monophyly of infraorder Scarabaeiformia within Polyphga. However, the Scarabaeiformia represented by three newly sequenced species was unexpectedly placed as a sister group to the Cucujiformia, instead of the placement to the sister to the Cucujiformia and Elateriformia group.

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.

There has been a substantial controversy on the phylogenetic relationships among butterfly families and several competing phylogenetic hypothesis have been suggested. Among them the relationships of (((Nymphalidae + Lycaenidae) + Pieridae) + Papilionidae) has been further widely accepted. In this study, we sequenced EF1-α, COI, and 16S rRNA from 62 species belonging to four true butterfly families, Papilionoidea. Phylogenetic analyses using BI, ML, and MP showed that the traditionally recognizable families were strongly supported as monophyletic groups, with the exception of Nymphalidae, wherein the singly included species of Danainae was placed as basal lineage of the Nymphalidae + Lycaenidae group. Phylogenetic relationships among families supported the sister group relationship of Nymphalidae and Lycaenidae strongly by all analyses and placed Papilionidae as the most basal lineage of the Papilionoidea. On the other hand, the relationships of Nymphalidae and Lycaenidae group to Pieridae were either unresolved, revealing trichotomy, or the relationships of (((Nymphalidae + Lycaenidae) + Pieridae) + Papilionidae) as previously supported by several morphological and molecular works supported. Detailed within-family relationships among some genera also are shown in the presentation.

whitefly, Bemisia tabaci (Gennadius) have a wide host range including cucumber, tomato, and pepper, resulting in loss of crop yield. In this study, we tested larvicidal efficacy of several on-the-market environment–friendly agricultural materials (EFAM) to select the effective products after the target pests were stabilized in indoor rearing condition. The developmental periods of two whiteflies are as follows: in the case of T. vaporariorum, egg duration is 9.6 days, and nymph is 18.9 days, and in the case of B.tabaci, egg durationis 7.4 days, and nymph is 15.2 days under 25℃ with relative humidity (RH) of 60±5% and a photoperiod of 16L : 8D. The total period of T. vaporariorum as 5 days longer than B. tabaci. Among 22 EFAMs six products showed more than 60% of insecticide efficacy for against T. vaporariorum BTVB, BTVD, BTVG, BTVL, BTVM, and BTVS. On the other hand, seven EFAM products including showed over 60% of insecticide efficacy against B. tabaci BTVD, BTVG, BTVK, BTVL, BTVM, BTVN, and BTVU. In the case of Spodptera litura previously, xxEFAMs were tesed against 2nd instar S.litura, and EFAMs were found to have more than 90% efficacy. Test of these six EFAMs against entire larval stages were performed in this study. Although some of these products showed still more than 90% of insecticidal efficacy against up to 3rd instar larvae, the efficacy of these EFAMs sharply decreased as ages increase, result is less than 60% of efficacy of the products at most. This result indicates the difficulty to control S. litura with the on-the-market EFAMs alone under economic injury level. Collectively, it is required to find more EFAMs and find alternative method to control those insect pests tested in this study.

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.