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).

We examined the effects of cadmium exposure and various temperature stress on the expression of Pardosa astrigera heat shock protein 70 (HSP70). To do this, P. astrigera HSP70 gene was cloned and its sequence determined. Female spiders collected from non-contaminated region were exposed to 40mM CdCl2 for 2, 4 and 6 weeks by dietary uptake. At the end of every 2, 4 and 6 weeks of exposure, a batch of 5 spiders was collected and total RNA was extracted from each batch of whole bodies. Female spiders were also exposed to different temperatures (20, 25, 30 and 35℃) for 3h and RNA extracted likewise. Transcription profiles of HSP70 in response to cadmium and temperature were determined by quantitative real-time PCR using 18S rRNA as reference gene for data normalization. HSP70 transcription gradually increased during 2,4 and 6 weeks of exposure to cadmium. In particular, the expression level at 6-week exposure was 3.4-fold higher than that of untreated control. In the temperature response, an increased expression of HSP70 was also observed as temperature increased up to 30℃ and then slightly decreased at 35℃. The expression level at 30℃ was 2.3-fold higher than that of 25℃. Taken together, HSP70 gene appears to be up-regulated by general stress factors including cadmium exposure and temperature increase.

A thioredoxin peroxidase (TPx) gene was cloned from the bumblebee, Bumbus ignitus. The B. ignitus TPx (BiTPx) contains an open reading frame of 585 bp encoding 195 amino acid residues and possesses two cysteine residues that are characteristic of 2-Cys subgroup of peroxiredoxin family. The deduced amino acid sequence of the BiTPx cDNA showed 90% identity to Apis melifera (AmTPx-1), 80% to Aedes aegypti (AaTPx), and 78% - 47% to other insect 2-Cys TPx. Northern blot analysis revealed the presence of BiTPx transcripts in all tissues examined. Western blot analysis showed the presence of the BiTPx in the fat body, midgut, muscle and epidermis, but not in the hemolymph, suggesting the BiTPx is not secretable. The cDNA encoding BiTPx was expressed as a 27-kDa polypeptide in baculovirus-infected insect Sf9 cells. The purified recombinant BiTPx was shown to reduce H2O2 in the presence of electrons donated by dithiothreitol and shown to be active in the presence of thioredoxin as electron donor.

Insect nicotinic acetylcholine receptors (nAChRs) are targets for insecticides. Despite the importance of the nAChR as a major target for insecticide action, modulators of nAChRs in insects remain unidentified. Here we describe the cloning and identification of a nAChR modulator gene in an insect. This gene was isolated by searching the firefly Pyrocoelia rufa cDNA library, and the geneitself encodes a protein 120 amino acids in length, named Pr-lynx1. Pr-lynx1 shares all the features, including a cysteine-rich consensus motif and common gene structure, of the Ly-6/neurotoxin superfamily. The recombinant Pr-lynx1, which is expressed as a 12-kDa polypeptide in baculovirus-infected insect Sf9 cells, is normally present at the cell surface asa GPI-anchored protein. Northern and Western blot analyses revealed that Pr-lynx1 is expressed in various tissues, such as the ganglion, brain, mandibular muscle, proventriculus, leg muscle, and epidermis. This expression pattern is similar to the distribution of nAChRs as assayed by α3 nAChR immunoreactivity. Co-expression of Pr-lynx1 in Xenopus oocytes expressing α3β4 nAChRs results in an increase in acetylcholine-evoked macroscopic currents, indicating a functional role of Pr-lynx1 as a protein modulator for nAChRs. This study on Pr-lynx1 is the first report of a modulator of nAChRs in an insect species.

The pinewood nematode (PWN, Bursaphelenchus xylophilus) causes the pine wilt disease, transmitted to pinewoods by the pine sawyer beetle, Monochamus alternatus. It is very difficult to discriminate B. xylophilus from B. mucronatus. Therefore, it has been necessary to detect PWN-infected trees for the prevention of pine wilt disease transmission in a short time. The development of biomarkers such as DNA and protein is important for diagnosis of B. xylophilus. However, there have been no reports regarding biomarker identifications of B. xylophilus. In this study, polyclonal antisera were raised against whole proteins of B. xylophilus in BalbC mice and were primarily screened with ELISA. Twenty five among over 500 cell lines releasing polyclonal antisera revealed B. xylophilus-specific immuno-reactivity, which was at least three times higher than that of B. mucronatus. Three cell lines among them were secreting monoclonal antibody through further screening. These cell lines only detect about a 33-kDa protein in B. xylophilus in the western blot. These results suggest that these monoclonal antibodies will be useful for the development of diagnostic kit for the pine wilt disease.

This study was performed to investigate the change of probing and feeding behavior of Q type of B. tabaci according to the decrease of residual effect of two insecticide, emamectin and pyridaben, using EPG technique. Examined the residual activity during 20 days, pyridaben is showed longer than emamectin benzoate. When treated two insecticides onto tomato leaves with the recommended concentration through 20 days, EPG waveforms of Q type of B. tabaci was recorded during three hours compare with the characteristic patterns of feeding behavior between two insecticides such as duration of first probing time, total duration of non-probing phase, total duration of probing phase and total duration of phloem phase. Recorded result from the change in total duration of probing activity to react the two insecticides, pyridaben was showed higher the time of probing activity, however, total duration of phloem phase was appeared low activity. Total duration of phloem phase with passage of days did not show until seven days, however, and gradually increased in emamectin benzoate after 10 days. In conclusion, residual effect between two insecticides was showed more rapidly decreased in emamectin benzoate, however, feeding behavior of Q type of B. tabaci was increased.

A B. thuringiensis kurstaki was first discovered by H. Dulmage in 1970 and commercialized as a DipelTM due to powerful toxicity to various Lepidoptera. Previously we isolated B. thuringiensis kurstaki on the basis of plasmid DNA profiling and H-antigen serotyping. The aims of this study were to screen larvicidal activities and select the highly active B. thuringiensis isolates against the important polyphagous pests of mandarine oranges and vegetables. The colony forming unit (CFU/ml) of each of culture mixtures was determined to estimate the δ-endotoxin concentration. The bioassay against artificial diet-rearing insects was conducted by surface contamination methods using the Petri dishes specially designed and manufactured by SPL Lifesciences. The insecticidal activities to the natural diet-rearing insect were measured by the application of spore and crystal mixtures to the leaf discs of the chinese cabbage with Potter spray tower. The following insects were used for the larvicidal activities of B. thuringiensis isolates: beet armyworm Spodoptera exigua, diamondback moth Plutella xylostella, giant looper Ascotis selenaria, tobacco cutworm Spodoptera litura, and variegated cutworm Peridroma sucia.

Bt crystal proteins, encoded by cry genes, are a group of insecticidal proteins unique in the Gram-positive and spore-forming bacterium, Bacillus thuringiensis. These cry genes are widely applied as one of the most successful candidates for constructing transgenic plants resistant to pest insects. In our previous report, we found Cry1-5 had high insecticidal activity against Spodoptera larvae although its amino acid sequences showed high similarity (97.9%) to those of Cry1Ab which had low activity. In comparison with Cry1Ac, Cry1-5 had 12 different residues in domain Ⅰ and domain Ⅱ, and we focused on domain Ⅰand domain Ⅱ regions and designed 10 mutagenic primers to change 12 residues. Through multi site-directed mutagenesis, we mutated the modified cry1Ac gene by plant codon usage in pOBⅠ-Mod-cry1Ac based on cry1-5 and constructed 63 various mutant cry genes. In the further study, we will express those mutant proteins as a fusion form with polyhedrin using baculovirus expression system and subsequently do bioassay to Spodoptera larvae.

A strain of Bacillus thuringiensis, named Bt 1-3, was isolated from Korean soil sample and it showed high insecticidal activity against Plutella xylostella. Bt 1-3 was deterimined to belong to ssp. aizawai (H7) by an H antiserum agglutination test and produced bipyramidal-shaped crystal proteins. PCR analysis with specific cry gene primers showed that Bt 1-3 contained cry1Aa, cry1Ab, cry1C, cry1D and cry2Ab genes. In addition, this isolate showed high uptake rate of foreign plasmid by electroporation. Based on these characteristics of Bt 1-3, we tried to construct a spore-free Bt 1-3 mutant by knock-out sigG gene, which is known as a key transcription factor during sporulation. First, we constructed a basal vector, named pDST, consisting of erythromycin resistant gene (EmR), partial polyhedrin gene and temperature sensitive origin of replication gene (Orits). Subsequently, according to the chromosomal DNA sequence of Bt subsp. konkukian 97-27, we amplifed upstream and downstream regions of Bt 1-3 sigG, and cloned into pDST (pDST-G). So far, several EmR colonies were obtained by electroporating into the wildtype Bt 1-3 and crossover by homologous recombination is going on.

The complete genomic nucleotide sequence of the Spodoptera litura multicapsid nucleopolyhedrovirus (SlMNPV) isolated in Korea, SlMNPV-K1, was determined. It was 137,435 bp long, with a 55.4 % A+T content and contained 132 putative open reading frames (ORFs) of 150 nucleotides or larger that showed minimal overlap. The 132 putative ORFs covered 87.7% of the genome. Among these, 131 ORFs were are homologous to genes identified in previously reported SlMNPV genome which consisted 139,342 bp and contained 141 putative ORFs. However, arrangement of some ORFs were somewhat different from each other. Even though the SlMNPV-K1 genome is smaller than that of previously reported SlMNPV genome and had lesser predicted ORFs, the main functional genes were all conserved. When the phylogenic relationship was analyzed using the nucleotide sequence of polyhedrin gene, SlMNPV-K1 was most closely related to Lymantria dispar multicapsid nucleopolyhedrovirus (LdMNPV) which were belonged to Group Ⅱ nucleopolyhedrovirus.

To develop an improved baculovirus insecticide with additional advantages, a novel recombinant baculovirus, AcB5B-AaIT was constructed. B. thuringiensis crystal protein gene (cry1-5) and insect-specific neurotoxin gene (AaIT) were introduced into Autographa californica nucleopolyhedrovirus genome by fusion of polyhedrin-cry1-5-polyhedrin under the control of polyhedrin (polh) gene promoter, and AaIT under the control of early promoter of ORF3004 from Cotesia plutellae bracovirus, respectively. About 150 kDa of Polyhedrin-Cry1-5-Polyhedrin fusion protein expressed by AcB5B-AaIT was occluded into the polyhedra produced by the recombinant virus, and activated as about 65 kDa of crystal protein when treated with gut-juice of Bombyx mori. The AcB5B-AaIT showed about 50% reduced LT50 value compared to that of the recombinant virus, Ap1Ac, expressing Cry1Ac against Plutella xylostella larvae. In addition, Spodoptera exigua larvae fed the recombinant polyhedra of AcB5B-AaIT showed about 4 fold higher refusing diet effect compared S. exigua larvae fed the recombinant polyhedra of the recombinant virus, Ap1C, expressing Cry1C. AcB5B-AaIT could be transferred to wild-type baculovirus along with serial passage by the homologous recombination between two polyhedrin genes contained in polh-cry1-5-polh fusion protein gene. These results suggested that the novel recombinant baculovirus, AcB5B-AaIT, could be applied as advanced viral insecticide.

To understand geographic genetic variation of the species and relationships among populations of the bumble bee, Bombus ardens, we sequenced a portion of mitochondrial COI gene, which corresponds to "DNA Barcode" region (658 bp) from 101 B. ardens individuals collected over 11 localities in Korea. The sequence data were used to investigate genetic diversity within populations and species, geographic variation within species, phylogeographic relationship among populations, and phylogenetic relationship among haplotypes. A total of nine haplortpes were found, but they very close to each other (a maximum sequence divergence of 0. 304%). Summarized, overall moderate to low genetic diversity within populations and species was characteristic, concordant with the high potential to disperse of B. ardens in Korea. There was no clear regional subdivision was observed and relatively high rate of gene flow among localities and low FST was characteristic.

This study was conducted to investigated the distribution, ecological character and life cycle of Black Soldier Fly(BSF), Hermetia illucens. The distribution of BSF was defined in all parts of the country in Korea. Its main habitat was found to be areas near cattle sheds, manure sheds, living waste dump grounds, and food waste dump grounds. Observed characteristics of BSF by developmental stage may be summarized as follows: eggs were a long oval shape of 886.9±19.7 ㎛ in major axis and 190.1±9.7 ㎛ in minor axis; they were 24.0±1.6 ㎍ in weight. One adult insect laid 1001±247 eggs in quantity; days to hatch from eggs (27℃, 60% R.H.) were 81.3±12.5 days. Last instar larva ranged from 20.7±1.1 mm in size, the length of larval stage was approximately 15～20 days. Pupae exhibited red brown, 19.2±1.1 mm in size; pupal state lasted 15.5±1.4 days for female, 14.7±1.4 days for male, exhibiting the tendency of males having shorter period than females. Adult insects were sized about 13～20 mm and colored black. Mating started 2 days after emergence and was most active during the 3rd day. Mating mostly occurred between 10:00 and 16:00 during which light intensity is high. Laying eggs started 3 days after emergence and was most frequent during days 4～6. Time of laying eggs during the day was similar to copulation time, showing the highest laying rate between 10:00 and 16:00.

As a major component of cuticle wax, cuticular hydrocarbon plays key roles as a chemical cue among inter- and intra-species in some beetles like as pheromone and alleolochemicals in addition to the physical function. In this study, cuticular hydrocarbon was analyzed and compared with three species of beetles adult, Monochamus alternatus, M. saltuarius belong to the Monochamus sp., and belong to the same class, Moechophyta diphysis, a typical vector of pine wilt disease. The composition of cuticular hydrocarbon of three species of beetles adult did not show the difference between male and female in intra-species, however, showed the difference in inter-species. They also differed from carbon numbers in inter species as 25-32 in M. saltuarius, 25-35 in M. alternatus, and 23-31 in M. diphysis. Major constituent of M. saltuarius was analyzed as n-C25, 2 or 4 MeC26, 9-C25:1, n-C27, 4-MeC28, n-C29, (9,x)-,and/or (11,x)-diMeC29; those of M. alternatus were n-C25, n-C27, 4-MeC28, n-C29, (9,x)-, and/or (11,x)-diMeC29, 9-C29:1, n-C29, (9,x)-,and/or (11,x)-diMeC29; and those of M. diphysis were n-C25, n-C26, n-C27, 3-MeC27,9-C29:1, 11-MeC29. The contents of n-alkanes were as follows: M. saltuarius ≒ M. alternatus > M. diphysis. The contents of monomethylalkanes were as follows: M. diphysis > M. saltuarius ≒ M. alternatus. The contents of dimethylalkanes were as follows: M. saltuarius ≒ M. alternatus > M. diphysis. And the content of olefine in female were analyzed as follows: M. saltuarius > M. alternatus ≒ M. diphysis, and the male's contents were similar in three kinds of beetles.

Among the cultured products of Beauveria bassiana SFB-205 (KCCM 10892P), the supernatant showed the highest insecticidal activity against 2nd instars of Aphis gossypii (Aphididae) nymphs under glasshouse condition. The enzymes in the supernatant were confirmed as active materials, and the chitinase was finally selected as a QC factor for commercial production. However, the chitinase activity in the supernatant decreased by 11-folds due to the thermal stress at 50℃ for 2 h. To obtain thermal stability, the chitinase in the supernatant was adsorbed to a precipitable material and the pellet was freeze-dried (PCT/KR2007/005886). The adsorbent-A showed about 92.7% of harvesting efficiency which was higher than the other candidates. The chitinase activity of the freeze-dried powder was kept up about 82.0% of initial activity for the same thermal stress. Subsequently, an optimal formulation recipe was established to maximize long-term storage stability and efficacy. SFB-205 oil-based formulation was stable up to 18 months at room temperature. It showed 96.1% efficacy against 2nd instars of A. gossypii nymphs at 1 day after the treatment in the glasshouse. This novel approach can be a promising method to develop competitive biopesticies in the entomopathogenic fungi, even though it needs to be intensively studied.

This study was performed to investigate the feeding behavior of Bemisia tabaci B- and Q-biotypes using EPG technique against seven tomato and eight red pepper commercial varieties. EPG waveforms was recorded during three hours compare with the characteristic patterns of feeding behaviors between two biotypes such as total duration of non-probing times, the time taken until stylet activities changed after reaction, number of probes, and total duration of phloem phase. In comparing the effect of the varieties between the two biotypes, biotype Q showed the feeding behavior against all pepper varieties in the total duration of phloem feeding. Daeshin variety has the longest feeding time. However in total duration of phloem feeding, biotype B was observed in hanyeoreumbigarim and Daeshin varieties, but feeding time was very short. Biotype B was longer in total duration of non-probing, and showed lower number of probes, and total duration of probing phase. However, biotype Q was shorter time in total duration of non-probing phase than biotype B, but showed more aggressive probing and stylet pathway pattern. These results suggest that biotype Q are more preferred the pepper host than biotype B. However, tomato varieties between the two biotypes did not show the difference.

Taxonomic studies of Taiwan moths have been done by only a few Taiwanese and foreign entomologists. The earliest significant collections and studies of Taiwan Lepidoptera began in 1856 with the arrival of Robert Swinhoe (1836-1877) as British Consul in Taiwan. The Japanese Occupation (Sino-Japanese War of 1895-1945) enabled studies by mainly Japanese scientists, and post- 1945 many Japanese and some others scientists visited Taiwan to study and collect moths and butterflies. In 1992, Heppner and Inoue (eds.) reported over 3,900 species in the checklist of "Lepidoptera of Taiwan" for the first time, based on the materials from 62 collecting sites and previous records. Recently,the Lepidoptera series of guide books to Insects in Taiwan was published by Mr. H.Y. Wang. Also the 2nd author has a publication plan of a total of 10 volumes in the series of "Lepidoptera of Taiwan", and a color introduction due out this year. In this study, we identified over 3,000 specimens of the subfamily Olethreutinae belonging to the family Tortricidae, collected at 62 sites in Taiwan from 1982 to 2006 by the 2nd author and some of collection’s of Y.S. Bae and K. T. Park in 1996-1999. We recognize the results of identification that more than 280 Taiwan tortricids, including over 35 newly recorded or new species, are in the Taiwan fauna.

Four species in two genera of the tribe Pilophorini, the ant mimetic Plant bugs are revised from Nepal. One new species of genus Pilophorus Hann, P. sp. nov is proposed. Pilophorus typicus (Distant, 1909) and Sthenaridea piceonigra (Motschulsky, 1863) are new record to Nepal. Pilophours josifovianus Duwal and Yasunaga, 2008 previously described species from Nepal and all other three species are here with illustrations of male and female genitalia and their biological information.

Many female moths produce and release sex pheromones to mate successfully with a conspecific male. Sex pheromone production in lepidopteran moth is known to be under the regulation of a pheromone biosynthesis activating neuropeptide (PBAN). A PBAN polypeptide is processed into five neuropeptides (NPs) after post-translational modification, resulting in diapause-like NP, PBAN, α-, β- and γ-NP. All of the peptides are amidated in their C-termini and shared a conserved motif, FXPR(or K)L structure. PBAN (Plx-PBAN) from Plutella xylostella consists of 30 amino acids, the shortest PBAN so far reported. When female adults were injected with synthetic Plx-PBAN, pheromone production showed a maximal increase 1 h post-injection. RT-PCR screening revealed that Plx-PBAN cDNA was expressed in both sexes, with the highest expression level in the head of female adults. Plx-PBAN binds to its receptor on pheromone gland cells. PBAN receptor has seven transmembranes, indicating G-protein coupled receptor family and transduces its signal via G-protein mediated signal transduction. Subsequently, calcium channels remain activated and stimulate biochemical reactions for sex pheromone production in the pheromone gland.