In order to identify the specific antigens for pine wood nematode (PWN), we confirmed that one of the genes commonly found in the transcriptome, proteome and secretory proteins of PWN belonged to the Aldose Reductase (AR) family protein. 36.5 kDa PWN-AR1 was expressed and purified using Baculovirus Expression System. Total 1,546 hybridoma fusion library was generated and screened for specificity to PWN-AR1 by Enzyme Linked Immunosorbent assay (ELISA). Nine clones showed strong immunoreactivity to PWN-AR1 were limited-diluted. Total 864 limited-diluted clones were further screened using PWN-AR1 by ELISA and 34 monoclonal antibody (Mab) clones were selected. 34 Mab clones were further screened using PWN extracts and a standard PWN-infected pine tree extract by ELISA. Finally nine clones were selected and their immunoreactivities to 4 different nematodes were examined by ELISA. Seven clones pecifically recognized PWN while two clones recognized 4 nematodes. Our data suggested that PWN-AR1 is a PWN secretory enzyme while PWN is invading pine trees, Thus, PWN-AR1-Mabs could be used to develop diagnosis tools for PWN and its infected pine trees. (This work was supported by National Institute of Forest Science)

To identify the venom components and their expression patterns of some Aculeata bees/wasps, venom gland-specific transcriptome analysis was conducted. FPKM values were normalized with the average of the transcription level of reference gene (a-tubulin). Common components in both solitary and social wasp venoms include hyaluronidase, phospholipase A2, metalloendopeptidase, etc. Although it has been expected that more diverse bioactive components with the functions of prey inactivation and physiology manipulation are present in solitary wasps, the information on venom compositions of solitary wasps obtained in this study was not sufficient to generalizae this notion. Nevertheless, some neurotoxic peptides (e.g., pompilidotoxin and dendrotoxin-like peptide) and proteins (e.g., insuline-like peptide binding protein) appear to be specific to solitary wasp venom. In contrast, several proteins, such as venom allergen 5 protein, venom acid phosphatase, and various phospholipases, appear to be relatively more abundant in social wasp venom. In the venom gland trancsriptome of bumblebees, major allergens or pain producing factors were barely identified, implying that bumblebee venoms are relatively less toxic than those of social or solitary wasps.

One of the leading pests of rice, small brown planthopper (Laodelphax striatellus) can grow up to have either short or long wings, depending on conditions. However, under the same breeding conditions, the phenotypes of the long- and short-winged small brown planthopper observed to keep the first collected phenotype. To investigate the mechanism involved in wing dimorphism, metabolomic researches have been conducted. In this study, we observed several metabolites change, and the difference of metabolites could provide clues to the relationship between physiological changes in the small brown planthopper and ecological transport.

한반도에서의 기후 변화는 최근까지 해충이 아니던 토착 곤충이나 비례 곤충들이 해충화 될 수 있는 가능성을 높여주고 있다. 가장 중요한 예가 토착 곤충이던 풀무치의 대량 발생에 따른 해충화가 있다. 낮은 밀도에서는 식생에 위험을 주지 않는 단독형으로 존재를 하던 풀무치가 평균온도의 상승과 강우 패턴의 변화로 대량 발생을 하여 강력한 이동성을 가지고 식생을 파괴하는 군집형으로 변화된 예가 있다. 하지만, 아직까지, 단독형에서 군집형으 로의 변화에 대한 기전이해는 완성이 되지 않은 상태이다. 기후의 변화는 겨울철 평균기온의 상승을 가져와서 한반도에서 월동이 불가능했던 해충이나 약제 저항성 계통들이 급속도로 번식할 수 있는 가능성을 열어 놓았다. 그러므로, 이러한 약제 저항성과 기후변화와 관련성에 대한 유전자 수준에서의 변화에 대한 이해가 필요한 실정이다. 위의 목적을 달성하기 위하여 우리는 다중분석학을 이용하여 연구를 진행하여 왔고, 연구 결과를 바탕으로 종 특이적이면 환경 친화적인 방제방법의 개발 연구를 진행하여 와서 이에 대한 연구결과를 발표한다(RDA Grant No. PJ010821032018).

소나무재선충(pine wood nematode, PWN) 감염 소나무를 현장에서 신속하게 진단을 할 수 있는 방법은 현재 없다. 본 연구에서는 PWN특이적인 항원으로 알려진 PWN-GaLectin을 baculovirus발현체계로 발현시켜서 총 1,464개의 fusion hybridoma 세포 주 라이브러리 를 제작 하는 항원으로 이용하였다. 총 1,464개의 fusion hybridoma 세포 주 중, PWN-GaLectin에 대한 높은 반응을 보이는 62개의 fusion hybridoma 세포 주를 선별했다. 이들 중, 표준 PWN 감염소나무 PBS추출물과 PWN 단백질 추출물에 강한 반응을 보이는 세포 주 12개를 선 별하여 단클론 항체(monoclonal antibody, Mab) 분비세포 주 확립을 위한 limited dilution을 실시하였다. Mab분비세포 주 확립을 위해서 표 준 PWN 감염소나무 추출물에 대한 반응이 표준 정상 소나무 추출물 보다 높은 세포 주들을 선별했다. 그리고 추가로 PWN 단백질 추출물에 대 해서 3종의 비 병원성 선충 단백질 추출물 보다 높은 반응을 보이는 세포 주들도 선별, 확립했다. 본 연구에서 확립된 Mab들을 우리는 현장과 실 험실에서 사용할 수 있는 신속진단키트의 개발에 이용할 수 있을 것이다.

풀무치의 전국적인 발생현황 및 밀도조사의 결과, 한국에서는 전라남도 해남군 산이면과 전라남도 무안군 망운면 간척지에서 2015년 이후 지속적으로 높은 밀도의 발생이 관찰되었다. 우리는 두 지점에서 발생하는 풀무치의 기원을 알아내기 위하여 NADH dehydrogenase subunit (NAD) 2, NAD4 와 NAD5의 염기서열을 분석하였다. 그 결과 해남풀무치의 경우는 중국동북부의 Liaoning성 과 Heilongjiang성 개체군과 기 원이 비슷하고, 무안풀무치의 경우는 일본풀무치와 기원이 비슷하다는 결론에 도달했다. 이전의 전 세계적인 풀무치의 진화에 관한 연구에서 한 국의 풀무치가 포함이 되지 않아서 한반도 풀무치의 기원은 알 수 없었다. 본 연구의 결과는 중국북동부 지방에서 8만 년 전에 분리된 풀무치 중 일부가 한반도로 이동을 하여 해남 지역에 정착을 하고 일부는 러시아 사할린과 일본 홋카이도섬을 거쳐서 무안으로 이동하였을 가능성을 보여주 고 있다. 하지만, 한반도로 내려온 풀무치가 해남과 무안계통으로 분리된 후 일본으로 이동하였을 가능성도 배제할 수 없다.

Steamed and freeze-dried mature silkworm powders (SMSP) have various health improvement effects. However, westill do not know which substances are reponsible for varius health improvement effect yet. In this study, we comparedcontents of phytochemicals in SMSP to mulberry leaves and other silkworm powders. We found that SMSP have certainlevels of phyttochemicals and silk proteins. Our data suggested that various substances in SMSP are responsible for healthimprovements effect. (Project title: Elucidation the health improvement effects of boiled silk worm larvae, Project No:PJ010828012017)

Biological activities of bombolitins from Bombus ardens, B. consobrinus, B. terrestris and B. ussurensis (bombolitinsA, C, T and U, respectively) were examined using hemolytic, anti-microbial, anti-fungal and anti-tumor activity assays.Among the four bombolitins tested, bombolitin T showed the highest hemolytic and anti-tumor activities. All bombolitinsexhibited strong anti-microbial and anti-fungal activities, and bombolitin A specifically possessed the highest anti-microbialactivity against the Gram-negative bacteria Escherichia coli. Circular dichroism spectrometry analysis revealed that allfour bombolitins had over 61.7% and 45.5% of α-helicity in 30 mM sodium dodecyl sulfate and 50% trifluoroethanolbuffers, respectively, which form lipid-membrane-mimicking environments. Bombolitin T showed the lowest IC50 valuesof 8.5 μM and 8.8 μM against SK-OV-3 and NIH-OVCAR-3 cell lines, respectively, after 72 h of treatment, but itsrelative hemolytic activity at a concentration of 200 μM was 2.3-fold higher than that of 0.1% Triton X-100. Thisstudy provides new information on the biological and molecular properties of venom peptides of bumblebees. However,further studies on reducing cytotoxicity of bombolitins are needed for designing selective anti-tumor peptides.

Disheveled is a major regulator of WNT signalling pathway. It has been shown that WNT signaling is important for regulating synaptic plasticity. However, it is not still clear how Dsh is regulating synaptic plasticity. In this study, we used various methods to investigate how Dsh regulates synaptic plasticity. Our further studies will reveal unknown molecular and cellular mechanisms underlying WNT signaling dependent synaptic plasticity.

Pine wilt disease is currently the most deadly forest disease in the world and is known to be caused by infection of Bursaphelenchus xylophilus. Until now, no method has been developed to confirm the pine tres infected with pine wood nematodes. In order to develop a method to diagnose pine wood nematode infection, we produced a monoclonal antibody against Expansin B3, which was found to be secreted by pine wood nematodes. ELISA assay using various monoclonal antibody confirmed that the pine trees infected with the rewarming can be detected. Further studies using our antibodies may help to develop a diagnostic method that can quickly confirm infection of pine wood nematodes in the field.

To investigate unknown molecualr and cellular mechanisms underlying polyphenisms in the migratory locusts, we performed whole transcriptome analysis from 12 different groups. Significantly differentially expressed coding and long non-coding (LNC) RNAs were present according to habitats, phases, and genders. Those differentially expressed coding and LNC RNAs were confirmed by quantitative-realtime-reverse transcriptase PCR. Those identified coding and LNC RNAs could be a target for developing environmental friendly management tools in the near future.

Polyphenism is one of the most intriguing phenomenon observed from various insect species. Polyphenism is changing morphological, behavioral, and physiological phenotypes without changing genome informations. Therefore, polyphenism is thought to be controlled by epigenetic methods. Epigenetic controls are including DNA methylation, histone acetylation, small RNAs, long non-coding RNAs, and protein phosphorylation. To investigate the mechanism involved in polyphenism, various genomic researches have been completed. However, metabolite changes in different phases have not been reported yet. Thus, we studied the metabolite changes and found the changed metabolites. These studies will make an important contribution to reveal the difference between gregrious and solitary phases and reveal eco - friendly control techniques.

Insect resistance to crop protecting chemicals is developing very rapidly, which is a major problem in the production of crops. Recently, the mechanism of resistance to various pseticides s has been revealed in Helicoverpa armigera. However, to date, no technology has been developed to quickly diagnose resistance yet. In this study, we present the results of developing method of rapid diagnosis of resistance

Abaecin is a proline-rich anti-bacterial peptide which is known to have high activity against Gram-negative bacteria and it is originally isolated from honeybee Apis mellifera. Since the anti-bacterial peptides including abaecin were identified in the hemolymph of A. mellifera infected with Gram-negative bacteria Escherichia coli, this result implies that honeybees express abaecin for defense against natural infection. Anti-microbial peptides including abaecin were also identified in the adult hemolymph of bumblebees. To investigate pharmacological and toxicological properties of abaecin from Bombus ussurensis, biological activities were evaluated by conducting anti-tumor, anti-microbial and cytotoxic activity assays. Abaecin exhibited significantly high anti-tumor activity against ovarian tumor cells SK-OV-3 and NIH; OVCAR-3 at 100 μM after 72 h treatment. Interestingly, abaecin showed none of anti-microbial activity against Gram-negative and Gram-positive bacteria. Since abaecin exhibited extremely low level of hemolytic activity against human erythrocytes, it may serve as a good model peptide for studying its potential as a selective anti-tumor drug.

Ryanodine receptors (RyRs) regulate the contractions of insect muscles by altering intracellular Ca2+ concentration and are the targets of chlorantraniliprole. Here we established two resistant strains of Drosophila melanogaster, which were treated with low or high concentrations of chlorantraniliprole, and their resistance levels were determined on the basis of contact and ingestion toxicities. Compared with the wild type, the two resistant strains did not show any significant differences in contact toxicity. However, they showed significantly increased resistance ratios in ingestion toxicity than that by the wild type. The resistant strains had altered expression levels of RyRs and more enhanced Acetylcholinesterase and Glutathione-S-transferase activities than that by the non-selected strain. These results suggested that the resistance development of chlorantraniliprole in the two strains might be mediated by the activation of detoxification pathways in D. melanogaster.

Maruca vitrata larvae feed on flowers and pods of several leguminous crops, and can severely reduce seed yield. Adult emergence, mating and oviposition behaviors were observed in a 15h/9h=light/dark and 25℃ condition as a base study for monitoring. Emergence occurred mainly during the first five hours of scotophase. Mating occurred from the two days after emergence, and the mated females started to lay eggs from the next day. A maximal mating rate was observed in the night of five days. Mating occurred significantly more often during the time from 2 hours before to 3 hours after lights-off, but older females mated more frequently during the photophase. Electroanntennographic responses of males to some chemicals, and body extracts and volatile collections of females were measured

We developed a simple molecular detection tool that rapidly and accurately identifies Spotted wing Drosophila (SWD) without sophisticated instruments or expertise. We first identified a gene that is present in the SWD genome but not in that of any other insect species. Then, we developed the loop-mediated isothermal amplification (LAMP) assay, which was designed based on genomic DNA of SWD specific gene. This LAMP assay detected only genomic DNA from SWD—not from Drosophila melanogaster. In addition, this assay could detect genomic DNA in SWD geographical strains collected from 8 different locations in Asia, Europe, Hawaii, and the USA. Our LAMP assay could be a useful detection tool for identifying SWD rapidly in the field. This work was carried out with the support of the Cooperative Research Program for Agriculture Science & Technology Development (Project No: PJ0116302016).

Bombolitin is a venom peptide originally isolated from bumblebees and possesses various biological activities, including hemolytic activity. Bombolitins exhibit amphipathic α-helical structure in lipid-membrane-mimicking environments. To investigate their pharmacological and toxicological properties, anti-tumor, anti-microbial and cytotoxic activities of bombolitins from Bombus ardens and Bombus ussurensis were evaluated. Bombolitins of the two species exhibited extremely high anti-tumor activity against ovarian tumor cells SK-OV-3 and NIH; OVCAR-3 at 25-50 μM, which is 2-fold more potent than other wasp venom peptides studied to date (Yoon et al., 2015; Yoon et al., 2016). The two bombolitins showed significantly high antimicrobial activity. However, bombolotin of B. ussurensis showed no antimicrobial activity against Escherichia coli. In addition to their high levels of anti-tumor activity, bombolitins showed considerable levels of hemolytic activity. Thus, to utilize bombolitins as a potential candidate for anti-tumor peptide drugs, further studies for reducing cytotoxic properties of bombolitns is essential.

The lesser paper wasp, Parapolybia varia, belongs to large subfamily Polistinae and is distributed in Middle East, the Indo-Papuan region and East Asia. P. varia is known to become aggressive when disturbed for defending their colonies, resulting in fatal envenomation. Vespid chemotactic peptide (VCP) and vespakinin have recently been determined to be the top two genes most abundantly transcribed in venom glands of P. varia. To investigate the pharmacological and toxicological properties of VCP and vespakinin, their antitumor, antimicrobial, and cytotoxic activities were evaluated. VCP exhibited a significantly high antitumor activity against ovarian tumor cell SK-OV-3 at 100 M. VCP also showed higher hemolytic activity than vespakinin. Antimicrobial activity was only observed with VCP against yeast Candida albicans at 1 mM. Since VCP showed a relatively low hemolytic activity but a considerable level of antitumor activity, it has further merits to be exploited as a potential antitumor agent with reduced side effects on normal cells.

Repetitive applications of drugs to tumor tissues and animals induced resistance and/or tolerances which caused severe problem in agriculture and medicine. However, we still do not clearly understand the molecular and cellular mechanisms underlying development of resistance and tolerance to chemicals. Drosophila is one of the most widely used model for studying fundamental phenomena in sciences using its available genetic and genomic resources. To investigate unknown molecular and cellular basis of drug resistance development, we applied Drosophila with two different concentrations of a chemical after treating them with Ethyl methanesulfonate mutatgenesis. We found that flies treating with two different concentration of chemicals showed different susceptibility to a chemical. We have established two different lines showing different susceptibility to a chemical. We will use these lines to compare any differences in mRNA expression profiling and enzyme activities. (This work was supported by project title: Investigation on cross drug resistance mechanisms using Drosophila as a model (PJ010821032016) from Rural Development Administration).