The occurrence and damage of stink bugs were monitored at South Korea from 2007 to 2008. A total of 11 stink bug species were observed in paddy field and occurrence periods of stink bugs were significantly differed by species. In occurrence period and damage of stink bug feeding in paddy field, Scotinophara lurida occurred from June, Cletus punctiger, Eysarcoris aeneus, Rhopalus maculatus, Riptortus clavatus, E. ventralis, Stenotus rubrovittatus occurred from August and Pachygrontha antennata, C. schmidti, Trigonotylus caelestialium, Paromius exiguus occurred from September. When we were classified into two region with mountain and plain area cultivated rice, generally C. puntiger population was higher in mountain area than plain area, and E. aeneus was higher in plain area than mountain area. Also, stink bug species were higher in environment condition formed with a paddy field and a waterway overgrown with weeds. Control threshold for E. aeneus in paddy field was 1-2 individuals/heading stage, and then rate of pecky rice made to 5.0% while milk stage, dough stage and yellow ripe stage made to less than 5.0%.

Two cherry tomato plant cultivars (Lycopersicon esculentum Miller, cultivars ‘Koko’ and ‘Pepe’) were supplied with high (395 ppm), medium (266 ppm) and low (199 ppm) concentrations of nitrogen to determine the influence of nitrogen fertilization on development, cultivar preference and honeydew production by greenhouse whiteflies, Trialeurodes vaporariorum (Westwood) (Hemiptera: Aleyrodidae). The nitrogen, protein, andchlorophyll content of tomato leaves were higher in the high nitrogen supplied plants than in the medium or low nitrogen supplied plants, but the sugar content showed an inverse relationship. The developmental times of eggsand nymphs decreased as the nitrogen concentrations increased in both cultivars. The preference of T. vaporariorum was compared by counting the number of eggs deposited on leaves in choice and non-choice tests. In the non-choice test, no significant nitrogen treatment effects were observedbut the upper plant stratum was preferred for egg laying. In the choice test, there were significant main effects of cultivar and nitrogen concentration. T. vaporariorum laid eggs more on leaves of plants with higher nitrogen at the upper stratum. In both experiments, T, vaporariorum preferred the ‘Koko’ cultivar to the ‘Pepe’ cultivar. The honeydew production of T. vaporariorum nymphs increased with decreasing nitrogen treatment concentrations. The largest honeydew production was detected in the ‘Pepe’ cultivar grown at low nitrogen concentration. It is concluded that cultivar ‘Pepe’ had an advantage over ‘Koko’ in term of T. vaporariorum management program in tomato greenhouses.

The green rice leafhopper, Nephotettix cincticeps is an insect vector which transmits virus diseases of rice plant causing severe damage to the rice and its damage both quality and yield of rice. Survival rate of overwintering N. cincticeps in Alopeculus acquadis among 15 species of the winter crops and weeds was 37.8% and its most preferred. Density of N. cincticeps in environment-friendly agriculture area which cultivated Astragalus sinicus as a green manure crop in winter season increased in number wintering pass in Alopeculus acquadis and Astragalus sinicus as host plants, because of control of N. cincticeps was not enough in the previous year. Survival rate of N. cincticeps was higher mainly in habitation mixed with rice straw, Astragalus sinicus and Alopeculus acquadis as winter habitation. It warmed due to the effect of the keeping warmth inside of a rice paddy field and the bank around a rice field covered with rice straw or Astragalus sinicus. Also, habitation conditions were optimal for survival because of growing Alopeculus acquadis as the major host plant of N. cincticeps in that place.

Moths were caught by using two 22W UV black light traps per treatment during May, June, August and September in 2007 from our study sites. Our study areas of the Korea Long Term Ecological Research (KLTER) sites were Mt. Nam with two plant communities (Quercus mongolica, Pinus densiflora), Mt. Jiri with three plant communities (Q. mongolica, P. densiflora, Abies koreana), Mt. Wolak with three plant communities (Q. mongolica, P. densiflora, Q. variabilis), and Mt. Jumbong with t재 plant communities (Q. mongolica, P. densiflora). The purpose of this study was to compare the species diversity of major plant feeders, Lepidopteran species (moths) at each forest type of the regional KLTER sites. Overall, the total numbers of moth species we’ve collected from the all KLTER sites in 2007 were 670 species. For the three plant communities, Mt.Jiri (11 family, 259 species, 2372 individuals) was the highest site and Mt. Wolak (7 family, 401 species, 2243 individuals) was the next highest. For the two plant communities, Mt. Jumbong (14 family, 166 species, 1750 individuals) was highest and Mt Nam (4 family, 21 species, 142 individuals) was very few. The ordination analyses, Nonmetric multidimensional scaling (NMS) and cluster analysis, showed distinct clusters separating the assemblages of moth found at each site and each plant community of the KLTER sites (P<0.05 from MRPP). Therefore, we suggest that our sustainable monitoring will verify the distinct cluster with the forest type at each site with many replications.

We assessed the environmental risk of herbicide resistant transgenic rice (Protox) on non-target herbivore, grasshoppers (Oxya japonica japonica Thunberg). We conducted life-history experiments of grasshoppers with measuring their body weight, body length, eating amount, and feces amount between non-transgenic rice (nTR; Dongjin rice) and transgenic rice (TR; Protox rice) under laboratory conditions (Temp. 25Ð, R.H. 50-70%, Photoperiod L16:D8) in 2007. The growth of grasshoppers appeared to increase at each measuring date. We also compared the growth rate of grasshoppers between nTR and TR to examine the transgenic impact on the herbivore and we found there was no statistically signifi cant difference between the two plant types (P>0.05). We found that body weight and body length for grasshoppers were highly correlated at each of the two types of plants, nTR (0.962) and TR (0.960). The correlation of eating amount and feces amount of grasshoppers were higher nTR (0.830) than TR (0.782). The energy effi ciency of the grasshopper was not a signifi cant between nTR and TR (P> 0.05). But the molt timing of the grasshoppers for TR difference was faster than for nTR. Conclusively life-history of the grasshoppers but molt timing was not a signifi cant difference between nTR and TR. Therefore, we could conclude there was not any environment risk on herbivore from our result.

This study was performed to investigate the seasonal occurrence, developmental characteristics of each nymphal stages with different temperatures (20, 25, 30℃), longevity and fecundity of ussur brown katydid, Paratlanticus ussuriensis, damaging by outbreaks in the orchard areas of Bitan-ri, Yeongdong, Chungbuk. Paratlanticus ussuriensis occurred from late-March to late-August with peak of mid-May. Newly emerged nymphs appeared from March and do damaged fruit orchards with peak of mid-May when P. ussuriensis existed as 4th and 5th nymphal stages. P. ussuriensis adult occurred from early-June to mid-Aug. with peak of mid-July. Total density of P. ussuriensis was showed highest in mid-May. P. ussuriensis goes through nymphal stages to 7th nymph, the ovipositor began exposed to outside from the 4th instar and the body weight increased heavily from this stage and the wings were observed from 6th instar. Developmental period was longer as increased the nymphal stages. Sex ratio of collected insect was showed as 0.57; females more than males. As increased the temperature, developmental period was to be short. Preoviposition was also to be short as 5.0, 4.3, and 3.4 days at 20, 25, 30℃, respectively, and fecundity increased as 69.0, 87.1, and 104.3 at 20, 25, 30℃, respectively. Longevity of male and female at 25℃ was showed the longest with 35.7, and 32.9 days and showed the shortest with 30.1 and 28.1 days at 30℃, respectively. The difference of developmental period in male and female were showed longer in female without relation of temperature. The eggs laid were frequently distributed 3 to 4 cm from soil surface, and showed the behavior laying eggs intensively when early oviposition period.

Sex pheromone production in lepidopteran is stimulated and regulated by a pheromone biosynthesis activating neuropeptide (PBAN). A cDNA of PBAN receptor (Plx-PBANR) isolated from female pheromone gland of the diamondback moth (DBM, Plutella xylostella (L.) encodes 338 amino acids. Plx-PBANR has conserved biochemical motifs and 7 transmembranes, indicating it belongs to G-protein coupled receptor family. Plx-PBANR expression was only found in female pheromone gland, demonstrating that pheromone gland is the only molecular target of Plx-PBAN. Human uterus carcinoma (HeLa) was stably transfected with Plx-PBANR gene and its expression was confirmed by RT-PCR analysis. Plx-PBANR expressing cells increased level of Ca2+ influx when challenged with Plx-PBAN and Hez-PBAN from Heliothis zea. When RNAi fragment for PBANR was injected into pupae, suppression of PBANR expression was confirmed by RT-PCR and maintained for at least 2 days at post-emergence. Injection of RNA fragment into pupae for inhibition of Plx-PBANR expression also inhibited mating behavior, revealing that reproductive organ of the female has no spermatocyte and that there are no successful reproductive behaviors. These results suggest of PBANR expression affects the molecular biological events of PBAN and eventually suppresses mating behavior.

vitellogenin cDNA was cloned from the bumblebee Bombus ignitus. The cDNA encoding B. ignitus vitellogenin (BiVg) is 5473 bases long with an open reading frame of 1773 amino acid residues. BiVg possesses two consensus (RXXR/S) cleavage sites and has the conserved DGXR and GL/ICG motif near its C-terminus. The deduced amino acid sequence of BiVg cDNA showed significant similarity with hymenopteran Vgs (51% identity to Apis mellifera Vg, and 33-36% to other insect Vgs). The BiVg cDNA was expressed as a 200-kDa polypeptide in baculovirus-infected insect Sf9 cells. Northern and Western blot analyses showed the expression of BiVg in fat bodies of pupae and adults. In queens, the expression of BiVg was first detected in late pupal stage fat bodies, and secreted BiVg was also observed in late pupal stage hemolymph.

Since the ancient times the therapeutic application of honeybee venom (BV) is practised and persisted until the present days. Resistant bacteria are in emergence and some drugs no longer have an antimicrobial action. To purify the melittin known as antibacterial peptide, five major peptidergic subfractions were separated, purified and identified from the whole BV. We investigated the antibacterial activity of whole BV and purified melittin against Staphylococcus aureus by the minimum inhibitory concentrations (MIC) and the postantibiotic effect (PAE). The MIC of whole BV for S. aureus was 0.06 ㎍/㎖, respectively. The MIC of melittin was 0.06 ㎍/㎖ on S. aureus. The in vitro PAE of whole BV and isolated melittin were determined using E. coli and S. aureus. The PAE of whole BV against S. aureus were 3.45 h (1×MIC). The PAE of melittin against S. aureus was 4.35 h (1 × MIC). Also both whole BV and melittin killed S. aureus at 5 × MIC. The regrowth wasn't observed after 18 h. These results suggest that whole BV and melittin will be developed a novel antibacterial drug.

The Black Soldier Fly (BSF, Hermetia illucens) was widely distributed throughout Korea. This insect was mainly found in the vicinity of and in cattle sheds, manure sheds, living waste dump grounds, and food waste dump grounds. This fly is a kind of a beneficial fly because BSF adults do not go into houses, they do not regurgitate on human food, they do not bite, bother or pester humans in any way and they are not associated in any way with the transmission of disease. But their greatest attribute lies their ability to eat and digest raw waste. They can devour, for example, a large, raw, Irish potato and others in just a few hours. Unlike many other flies, since the BSF larvae have very powerful mouth parts and digestive enzymes, they can ingest raw waste far more efficiently than any other known species of fly. On this study, to investigate whether feeding strategy of the BSF larvae involves extra-oral digestion or not, and to better understand this process, the salivary glands and a few tissue from the BSF were dissected and subjected to morphological and preliminary enzyme characterization.

Varroa destructor is an ecto-parasite mite and worldwide pest of the honey bee Apis mellifera L. The pyrethroid tau-fluvalinate (Apistan), an acaricide that is tolerated by honey bees, has been used for varroa mite control since the mid 1980s. Even though various resistances to tau-fluvalinate in varroa mites have been reported from Europe, Israel, and USA, the nature of tau-fluvalinate resistance in varroa mites in Korea has never been investigated. To investigate and understand tau-fluvalinate resistance in varroa mites in Korea, we conducted bioassay in several apiaries located different regions in Korea. To understand the molecular mechanisms underlying the difference of tau-fluvalinate resistances in varroa mites, partial genomic DNA fragments of a voltage-sensitive sodium channel gene from varroa mites were cloned and sequenced, since tau-fluvalinate is known to act on the sodium channels directly. Two novel mutations in sodium channels of varroa mites were present in eight apiaries. Two mutations might be a geographical polymorphism of sodium channel of varroa mites in Korea.

Apolipophorin-Ⅲ (apoLp-Ⅲ) is a hemolymph protein whose function is to facilitate lipid transport in an aqueous medium in insect. Recently, apolipophorin-Ⅲ in Galleria mellonella and Hyphantria cunea was shown to play an unexpected role in insect immune activation. We show here a novel possible function/role of apoLp-Ⅲ in insects. To investigate the genes which have a relationship with apoLp-Ⅲ in fall webworm larvae, we reduction of endogenous Hc apoLp-Ⅲ mRNA levels in larvae via RNA interference (RNAi). The RNAi-mediated Hc apoLp-Ⅲ reduction resulted in the reduction of antioxidants, like MnSOD, catalase, and glutathione S transferase as well as immune proteins. In particular, expression of MnSOD commonly decreased in fat body, midgut, and hemocytes following the knockdown of Hc apoLp-Ⅲ, which induced an elevated level of superoxide anion in H. cunea larvae. The observed effect of Hc apoLp-Ⅲ RNAi suggests that Hc apoLp-Ⅲ is related to the action/expression of antioxidants.

Innate immunity responses are triggered by the immune challenge and therefore involve signaling processes. The cellular response is initiated by hemocytes and mainly involves phagocytosis and encapsulation of intruders by these cells. To address whether Hc-STAT is activated upon bacterial challenge, we examined the subcellular location of STAT protein in hemocyte by immunostaining. A new insect member of the STAT family of transcription factors (Hc-STAT) has been cloned from the lepidopteran, Hyphantria cunea. The domain involved in DNA interaction and the SH2 domain are well conserved. The gene is transcribed at a low level during all stages of development, and the protein is present in hemocytes, fat body, midgut, epidermis, and Malphigian tuble (Mt). Especially, hemocytes and Mt showed transcriptional activation of Hc-STAT upon Gram (-) bacteria and fungal challenge. Gram (-) bacteria and fungal challenge specifically results in nuclear translocation of Hc-STAT protein and induction of DNA-binding activity that recognizes a STAT target site in H. cunea hemocyte. In vitro treatment with pervanadate translocates Hc-STAT to the nucleus in hemocyte cells. Here we report the first evidence for the involvement hemocyte JAK/STAT pathway upon microbial infection in lepidopteran insect.

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.

Sweet potato whitefly Bemisia tabaci is a serious pest of various economically important crops. For the control of B. tabaci in an environment-friendly way, we demonstrated the effect of azadirachtin, which is an active conpound of neem oil as an botanical insecticide, on the development of B. tabaci by using an assay of single tomato leaf. Egg hatch rates were 53.6, 50.3% at 5 and 10 ppm, respectively. Adult eclosion rates were 30.0, 22.9% at these doses. We determined the whitefly control efficacies of two application methods of neem-based products by comparing between a direct spray of liquid-type into leaves and a soil treatment of pellet-type. Soil treatment of neem was greatly inhibited adult colonization by 75%. Those plants also inhibited the rates of oviposition and larval development. However, single treatment of foliar spray of neem (5-10 ppm) did not significantly inhibit the initial colonization of adult whiteflies. Furthermore, adult colonization was gradually increased regardless of neem spray.

A cDNA of PBAN receptor (Plx-PBANR) isolated from female pheromone gland of the diamondback moth (DBM, Plutella xylostella (L.) encodes 338 amino acids. Plx-PBANR includes 7 transmembranes, indicating it belongs to G-protein coupled receptor family. Plx-PBANR showed high similarities with other moth PBANRs and its expression was only found in female pheromone gland, demonstrating that pheromone gland is the only molecular target of Plx-PBAN. To accomplish the funcional expression of Plx-PBANR, Human uterus carcinoma was stably transfected with Plx-PBANR gene and Plx-PBANR expression was confirmed by RT-PCR analysis. Plx-PBANR expressing cells increased level of Ca2+ influx when challenged with Plx-PBAN and Hez-PBAN from Heliothis zea, as ionomycin as a positive control does. To inhibit Plx-PBNAR expression in vivo, RNAi fragment for Plx-PBANR was injected into pupae. Suppression of PBANR expression was confirmed by RT-PCR and also induced inhibition of mating behavior in adults, revealing that reproductive organ of the female has no spermatocyte and that there are no successful reproductive behaviors. RNAi-treated adults showed reduced pheromone production. These results suggests that inhibition of PBANR expression affects the molecular biological events of PBAN and eventually suppresses mating behavior.

The root zone applications of a systemic insecticide, carbofuran, were evaluated for their impacts on the brown planthopper, Nilaparvata lugens (Stål), and spider populations in the greenhouse and rice paddy fields. In the green house experiments, no BPH nymphs were hatched at root zone treated on 40 to 50 day-old rice, while around 20 to 54 nymphs per pot were emerged in broadcasting and foliar spray treatments. This indicates that the root zone treatment can kill the eggs of BPH effectively. This is the first study ever demonstrated the high egg mortality of BPH due to the root-zone application. In the field experiments, the density of BPH in root zone treated plots were four to six times lower than in broadcasting and foliar spray plots at the 21 days after application. The BPH outbreaks and hopper-burns were observed at all treatments except the root zone treated plot at the 28 days after application. The root-zone application did not impact on the spider population, while foliar spray killed most of all spiders just one day after application. The densities of spider in foliar spray plots were always lower than in root-zone treated and control plots. The results indicated that the root-zone application of carbofuran can control BPH effectively without adverse effects to the spiders inhabited on the paddy field.

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.