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.

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 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.

The bumblebees, Bombus species are valuable natural resources being utilized for greenhouse pollination. Low level of genetic variation of Bombus species has been reported previously. In this study, we sequenced complete internal transcribed spacer 2 (ITS2) of the nuclear rDNA from 100 individuals of B. ardens collected from seven localities in Korean peninsula. The ITS2 sequence of B. ardens is longest, ranging in size from 1,940 bp-1,954 bp among known in insects, which ranges approximately from 241 bp-1,728 bp. The ITS2 sequences have ~51% of C+C content and contain each two 27 bp repeats, 20 bp repeats, 33 bp repeats, and 34 bp repeats at the beginning. Such repeats were not found in other insects. Uncorrected pairwise distance among 96 sequences that were obtained from 100 individuals revealed a maximum sequence divergence of 1.03%. Genetic diversity (π) of each population ranged from 0.007801 to 0.009627, and the lowest diversity was obtained from islet population of Ulleungdo, indicating possibly small, isolation of the population. Significant level of genetic distance was only found when Ulleungdo population was compared to two other mainland populations. Except for this, overall, a very high rate of per generation migration ratio (Nm=7.1-infinite) and a very low level of genetic fixation (FST=0-0.06546) were detected between pairs of localities. Analysis of hierarchical relationships among localities consistently revealed no statistically significant structure among populations. Taken these together, the B. ardens populations on the Korean peninsula are panmictic this is consistent with our understanding of the dispersal capability.

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.

Invertebrate mitochondrial genome contains 13 protein-coding genes and major start codons for them are ATA (Met) and ATG (Met). However, alternative start codons such as ATT (Ile), ATC (Ile), TTG (Leu), and GTG (Val) also have been suggested from a diverse organism. Approximately 120 complete mitochondrial genome reported showed that the start codon for COI gene evidences an array of diverse designation of COI start codon such as typical ATN, tetranucleotide TTAG and ATAA, newly proposed AAT and AAC and so on. In the case of Lepidoptera, many completely sequenced species showed no typical start codon at the start context of COI and even within the neighboring tRNATyr. In order to clarify, we newly sequenced the beginning context of COI gene, encompassing the neighboring tRNATyr and start region of COI gene from 39 species belonging to eight lepidopteran families. We found the newly sequenced 39 species and 14 available complete lepidopteran mitochondrial genomes all possessed CGA (arginine), which is the first non-overlapping in-frame codon in COI gene. Furthermore, this CGA is highly well aligned in terms of both nucleotide and amin o acid sequences with neighboring region. Thus, the CGA (arginine) may be synapomorphic character for Lepidoptera, functionally constrained. We, therefore, propose the CGA sequence as the start codon for COI gene in lepidopteran insects.

The bumblebee, Bombus ardens, is a valuable natural resource, and is most notably utilized for greenhouse pollination. In order to gain a greater understanding of the population genetic structure and the genetic diversity of this species, we sequenced a portion of the mitochondrial COI gene corresponding to the “DNA Barcode” region (658 bp) from 160 individuals collected over 15 Korean localities. Uncorrected pairwise distances among the eight haplotypes suggested low intraspecific genetic diversity, with a maximum sequence divergence of 0.3%. Such a low level of intraspecific genetic diversity was further reflected in local populations, particularly to islet populations, such as those of Youngheungdo, Jakyakdo, and Ulleugdo, which had zero genetic diversity. Geographically, one haplotype (BARBA01) was widespread and dominant, with a frequency of 90.6% (145 among 160 individuals). Other haplotypes were restricted to one to three localities and had low frequency. Overall, a very high rate of per generation female migration ratio (Nm= 4.6 to infinite) and a very low level of genetic fixation (FST= 0 to 0.099) were detected between pairs of localities, suggesting that the B. ardens population sonthe Korean peninsula are panmictic, which is consistent with our understanding of their dispersal capability.

We analyzed a portion of mitochondrial COI gene sequences (658 bp) to investigate the genetic diversity and geographic variation of the swallowtail butterfly, Papilioxuthus L., and the cabbage butterfly, Pieris rapae (Lepidoptera: Papilionidae). P. xuthus showed a moderate level of sequence divergence (0.91% at maximum) in 15 haplotypes, whereas P. rapae showed a moderate to high level of sequence divergence (1.67% at maximum) in 30 haplotypes, compared with other relevant studies. Analyses of population genetic structure showed that most populations are not genetically differentiated in both species. The distribution pattern of both species appears to be consistent with category IV of the phylogeographic pattern sensu Avise (Avise et al. 1987): a phylogenetic continuity, an absence of regional isolation of mtDNA clones, and extensive distribution of close clones. The observed pattern of genetic diversity and geographic variation of the two butterfly species seems to reflect the abundant habitats, abundant host plants, and flying abilities in connection with the lack of historical biogeographic barriers.

To understand geographic genetic variation of the species and relationships among populations of the bumble bee, Bombus ardens, that is utilized as green house pollinator we expanded our investigation by sequencing somewhat longer mitochondrial DNA (mtDNA) fragment, covering some uninvestigated regions within the species distribution, and analyzing the sequence data in terms of population genetic structure. For the purpose of study, a portion of mitochondrial COI gene, corresponding to "DNA Barcode" region (658 bp) was sequenced from 160 individuals of B. ardens collected over 15 localities in Korea. The sequence data revealed overall relatively low genetic diversity within species, with a maximum sequence divergence of 0.3%. Geographically, one haplotype (BARBA01) was found in all localities surveyed, with the frequency of 91% (145 among 160 individuals), whereas other haplotypes were found in a locality mostly as a single individual, suggesting that haplotype distribution can be summarized as coexistence of widespread, one dominant haplotype and regionally restricted, other haplotypes. Overall, very high rate of per generation female migration (Nm = 4.6 ~ infinite) and very low level of geographic substitution (FST = 0 ~ 0.099) among localities were characteristic. Although some populations were genetically subdivided from the remaining localities in the hierarchical analysis, there was regional polarity on this subdivision. Taken together with gene flow estimates, the nature of genetic divergence of the bumble bee populations is characterized as one that possessing low genetic diversity, high gene flow, and wide spread of one dominant haplotype, consistent with the previous finding. To have further detailed information of this valuable genetic resource, further longer and variable molecular portion is under investigating.

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).