Even though plant protections using chemical pesticides have several advantages, non-specific toxicities to other beneficial insects and humans and rapid development of tolerance and/or resistance of target pests to chemicals are major disadvantages. Recent researches suggested that using double stranded RNA (dsRNA) could be species specific and environmental friendly pest management protocols. However, efficiency of dsRNA treatments are known to be variable according to its application methods. For example, injections of dsRNAs to pests were known to be effective in all species. However, efficiencies of oral application of dsRNAs were known to be dependent on species. Thus, development of tools that could enhance the efficacy of orally treated dsRNAs are utmost important for widening usages of dsRNAs in plant protection. Recently, we found that the efficacy of oral treated dsRNAs to target pests could be enhanced by nano-technologies. I will show how applying nano-technology to dsRNAs enhance the efficiency of dsRNAs. (This work was carried out with the support of the Cooperative Research Program for Agriculture Science & Technology Development (Project title: Studies for biological characteristics of and control methods against the migratory locust, Locusta migratoria, Project No: PJ011630042016), Rural Development Administration).

Porcine epidemic diarrhea virus (PEDV) causes acute diarrhea and dehydration in pigs with high mortality. PEDV is belong to Coronavirus, enveloped, single-stranded, positive-sense RNA virus. PEDV particles were composed of four structure proteins such as a glycosylated peplomer (spike, S) protein, envelope (E), glycosylated membrane (M) protein, and unglycosylated RNA-binding nucleocapsid (N) protein. Many of previous studies talk about this four structure proteins have a great potential to diagnosis and prevent PEDV. In this study we investigated expression of these structure proteins using the bacterial and baculovirus expression system. In bacterial expression system, our results showed that structure proteins fused polyhedrin and intein gene were expressed higher than non-fusion structure proteins. The expressed fusion proteins were used to immune mice for generating a polyclonal antibodies. In baculovirus expression system, co-infection of insect cells with these four recombinant baculoviruses led to self-assembly of virus-like particles as demonstrated by Transmission electron microscopy. They were confirmed by western blot analysis using pre-made polyclonal antibodies. Finding in this study may provide important information for vaccine and diagnostic development.

Insects are among the most diverse groups of animals on the planet, representing more than half of all known living organisms. These insects are found in nearly every environment. Although humans regard certain insects as pests and attempt to control them using insecticides, most insects perform complex ecological functions, and provide either direct or indirect economic benefits to humans. Recently, the importance of insects used as food sources or as pets has increased in many countries, including Korea. In addition, several insects have a strong influence on people's emotion. Insect-mediated mental healthcare program is designed to help people who have disorders with physical, behavior and development. Children who have mental disorder, the experimental group that was provided with an insect-mediated mental healthcare program over a total of 8 sections, one section per week, 60 minutes per section, followed by pre-test and post-test. They responded to therapeutic effect after the completion of the program. Further research on the basis of this study is expected to help children with emotional therapy in other areas.

The population of managed honey bees has been dramatically declining the recent past in worldwide. The one of most common disease of bees is nosemosis, the nosemosis is caused by microsporidia in the genus Nosema. Nosema apis and N. ceranae have been described as honeybee pathogens. These microsporidia are highly evoloved fungi with an obligately intracellular parasitic lifstyle. The disease causes significant detriment to honey production and results in economic losses. In our knowledge, Fumagillin is the only antibiotic approved for control of nosemosis in honey bees, however this antibiotic may have unintended effects on the honey bee host, ultimately contributing to increased prevalence and pathogenicity of Nosema. Therefore, we screened anti-Nosema substances from entomopathogenic fungal culture filtrates using in vitro polar tube germination assay. These fungal metabolites are employed as antibiotic agents. As results, Total 3 samples (23% of 13 total samples) showing the germinating inhibition against N. ceranae. This screening method may be useful for the detection of anti-Nosema substances from various samples and selected samples in this study may be a good feature to be used in the development of a new biocontrol method of nosemosis.

백두대간은 한반도의 다양한 생물 서식지이며 동물의 이동통로로 새로운 서식지로의 이주와 재서식을 하는데 중요한 역할을 할 것으로 기대되고 있다. 본 연구는 2013-2014년에 4월부터 9월까지 백두대간 구룡령(명계리)과 고직령(서벽리)의 나비군집을 모니터링 하고, 두 지역의 나비를 과거기록과 비교하여 분포지의 변화가 있는지를 알아보았다. 그 결과 명계리 65종 1098개체, 서벽리 63종 1161개체가 조사되었다. 두 지역의 비교에서 명계리에서만 출현한 종은 15종, 서벽리에서만 출현한 종은 13종으로 나타났다. 종 순위별 개체수에서는 명계리는 큰줄흰나비 127 (11.6%), 줄흰나비 100 (9.1%), 산제비나비 90 (8.2%), 서벽리는 뿔나비 146 (12.6%), 줄흰나비 134 (11.5%), 네발나비 69 (5.9%)종으로 나타났다. 그리고 3개체 이하를 보인 종은 명계리 24종, 서벽리 23종으로 나타나 높은 비율을 차지하였다. 두 지역의 유사도는 비교적 높게 나타나고 있을 뿐만 아니라 한국적색목록의 나비가 다수 분포하고 있으며, 출현종을 과거 분포자료와 비교한 결과 뚜렷한 변화는 없었다. 이에 백두대간의 나비는 아직까지 잘 보전되고 있는 것으로 보여 거시적으로 백두대간의 서식지 경관이 연속성을 유지하도록 하는 것이 필요하다.

Porcine parvovirus (PPV), a member of the genus Parvovirus, family Parvoviridae, is a significant causative agent in porcine reproductive failure, causing serious economic losses in the swine industry. PPV is a non-enveloped virus and its capsid is assembled from three viral proteins (VP1, VP2, and VP3). The major capsid protein, VP2 is the main target for neutralizing antibodies in PPV. When VP2 was expressed in large amounts, it assembled into virus-like particles (VLPs) similar in size and morphology to the original virions. In this study, we generated the recombinant Bombyx mori nucleopolyhedrovirus (BmNPV) to express the VP2 protein. Expression of the VP2 protein was analyzed by SDS-PAGE and Western blot. The recombinant VP2 protein of approximately 64 kDa was detected by both analyses. The formation of VLP by recombinant VP2 was confirmed through transmission electron microscopy examination. The purified VP2 protein assembled into spherical particles with diameters ranging from 20 to 22 nm.

본 논문은 1982년과 2002년 사이 동북아시아 일부 대도시 지역을 대상으로 시계열 위성영상자료를 활용하여 도시팽창양상을 식별하는 것을 목표로 한다. 도시팽창은 시·공간적 역동성을 띄는 지표현상이나, 기존 연구에 주로 사용되었던 원격탐사 방법론들은 이의 시간적 측면보다는 공간적 측면에 비중을 두어 분석을 수행해왔다. 본 연구에서는 시계열 푸리에분석과 시계열회귀분석을 조합한 계량적 방법론을 이용하여 도시팽창의 시간적 측면에 보다 비중을 두고 분석을 하였으며, 동북아시아 대도시 지역에서 도시팽창이 어떻게 진행되는지를 지리정보체계/원격탐사 컴퓨터 프로그램인 이디리시를 사용하여 분석한다. 그 결과, 본 논문에서 시도한 방법론은 대도시 지역의 시·공간적 피복변화를 감지하는데는 효과적이었으나, 도시팽창 자체를 탐지하는데 있어서는 제약이 있는 것으로 나타났으며, 제약의 주된 이유는 도시지역의 피복과 토지이용의 대응이 항상 일치하지 않기 때문인것으로 파악되었다. 본 논문에서 제시한 시계열 원격탐사 방법론은 각 지역에서 관찰되는 각기 다른 도시화 양상을 픽셀단위로 구분하는데, 다양한 조합의 분석결과를 활용하면 도시팽창 연구에 유용할 것으로 예상된다.

Insect constitute the largest and most diverse group of animals on world and also serve as the hosts or nutrient sources. In addition, several insects have a strong influence on people's emotion. To utilize the preference and interest of insects in the field of mental healthcare, a survey study was conducted with individual living in Korea. As results, the most people had a high preference and interest of insect, but some were disagreeable to the insect itself. The preference and interest of insect were high on male, adult and practician experienced insect-related events than female, student and non-practician, respectively. The most favored insects were familiar or pet insects such as Papilio xuthus, Lucanus maculifemoratus, Allomyrina dichotoma and Lampyridae. These results may be useful to develop a healing program for mental healthcare using insects. Further research is needed to determine the effects of these insect in the mental therapy for this purpose.

Peptidoglycan recognition proteins (PGRPs) are family of innate immune molecules that recognize bacterial peptidoglycan. PGRP-LE, a member of the PGRP family, selectively binds to diaminopimelic acid (DAP)-type peptidoglycan to activate both the immune deficiency (IMD) and proPhenoloxidase (proPO) pathways in insects. A PGRP-LE-dependent induction of autophagy to control Listeria monocytogenes has also been reported. We identified and partially characterized a novel PGRP-LE homologue, from Tenebrio molitor and analyzed its functional role in the survival of the insect against infection by a DAP-type PGN containing intracellular pathogen, L. monocytogenes. The cDNA is comprised of an open reading frame (ORF) of 990 bp and encodes a polypeptide of 329 residues. TmPGRP-LE contains one PGRP domain, but lacks critical residues for amidase activity. Quantitative RT-PCR analysis showed a broad constitutive expression of the transcript at various stages of development spanning from larva to adult. RNAi mediated knockdown of the transcripts followed by a challenge with L. monocytogenes showed a significant reduction in survival rate of the larvae, suggesting a putative role of TmPGRP-LE in sensing and control of L. monocytogenes infections in T. molitor. These results implicate PGRP-LE as a defense protein necessary for survival of T. molitor against infection by L. monocytogenes.

Most traditional genome sequencing projects involving infectious viruses include culturing and purification of the virus. This can present difficulties as an analysis of multiple populations from multiple locations may be required to acquire sufficient amount of high-quality DNA for sequence analysis. The electrophoretic method provides a strategy whereby the genomic DNA sequences of the Korean isolate of Pieris rapae granulovirus (PiraGV-K) were analyzed by purifying it from host DNA by pulsed-field gel electrophoresis, thus simplifying sampling and labor time. The genomic DNA of infected P. rapae was embedded in agarose plugs, digested with a restriction nuclease and methylase, and pulsed-field gel electrophoresis (PFGE) was used to separate PiraGV-K DNA from the DNA of P. rapae, followed by mapping of fosmid clones of the separated viral DNA. The double-stranded circular genome of PiraGV-K encodes 120 open reading frames (ORFs), covering 92% of the sequenced genome. BLAST and ORF arrangement showed the presence of 78 homologs to other genes in the database. The mean overall amino acid identity of PiraGV-K ORFs was highest with the Chinese isolate of PiraGV (~99%), followed up with Choristoneura occidentalis ORFs at 58%. PiraGV-K ORFs were grouped, according to function, into 10 genes involved in transcription, 11 involved in replication, 25 structural protein genes, and 15 auxiliary genes. Genes for Chitinase (ORF 10) and cathepsin (ORF11), involved in the liquefaction of the host, were found in the genome. The recovery of PiraGV-K DNA genome by pulse-field electrophoretic separation from host genomic DNA had several advantages, compared with its isolation from particles harvested as virions or inclusions from the P. rapae host. We have sequenced and analyzed the 108,658 bp PiraGV-K genome purified by the pulsed field electrophoretic method. The method appears to be applicable to the analysis of genomes of large viruses. The chitinase, identified by PiraGV-K genome sequence, was functionally characterized by quantitative PCR, Western blot analysis, immunohistochemistry and transmission electron microscopy.

CD63, a member of tetraspanin membrane protein family, plays pivotal role in cell growth, motility, signal transduction, host-pathogen interactions and cancer. In this work, the cDNA encoding CD63 homologue (TmCD63) was cloned from larvae of coleopteran beetle, Tenebrio molitor. The cDNA is comprised of an open reading frame of 705 bp, encoding putative protein of 235 amino acid residues. In silico analysis shows that the protein has four putative transmembrane domains and one large extracellular loop. The characteristic ‘Cys-Cys-Gly’ motif and ‘Cys188’ residues are highly conserved in the large extracellular loop. Phylogenetic analysis of TmCD63 revealed that they belong to the insect cluster with 50-56% identity. Analysis of spatial expression patterns demonstrated that TmCD63 mRNA is mainly expressed in gut and Malphigian tubules of larvae and the testis of the adult. Developmental expression patterns of CD63 mRNA showed that TmCD63 transcripts are detected in late larval, pupal and adult stages. Interestingly, TmCD63 transcript was upregulated the maximum 4.5 fold in response to DAP-type peptidoglycan during the first 6 h, although other immune elicitors also made significant increase in the transcript level at later time-points. These results suggest that CD63 might contribute to T. molitor immune response against various microbial pathogens.

Apolipophorin III (apoLp-III) is a well-known hemolymph protein having a functional role in lipid transport and immune response of insects. We cloned full-length cDNA encoding putative apoLp-III from larvae of the coleopteran beetle, Tenebrio molitor (TmapoLp-III), by identification of clones corresponding to the partial sequence of TmapoLp-III, subsequently followed with full length sequencing by a clone-by-clone primer walking method. The complete cDNA consists of 890 nucleotides, including an ORF encoding 196 amino acid residues. Excluding a putative signal peptide of the first 20 amino acid residues, the 176-residue mature apoLp-III has a calculated molecular mass of 19,146 Da. Genomic sequence analysis with respect to its cDNA showed that TmapoLp-III was organized into four exons interrupted by three introns. Several immune-related transcription factor binding sites were discovered in the putative 5’-flanking region. BLAST and phylogenetic analysis reveals that TmapoLp-III has high sequence identity (88%) with Tribolium castaneum apoLp-III but shares little sequence homologies (<26%) with other apoLp-IIIs. Homology modeling of Tm apoLp-III shows a bundle of five amphipathic helices, including a short helix 3’. The ‘helix-short helix-helix’ motif was predicted to be implicated in lipid binding interactions, through reversible conformational changes and accommodating the hydrophobic residues to the exterior for stability. Highest level of TmapoLp-III mRNA was detected at late pupal stages, albeit it is expressed in the larval and adult stages at lower levels. The tissue specific expression of the transcripts showed significantly higher numbers in larval fat body and adult integument. In addition, TmapoLp-III mRNA was found to be highly up-regulated in late stages of L. monocytogenes or E. coli challenge. These results indicate that TmapoLp-III may play an important role in innate immune responses against bacterial pathogens in T. molitor.

We have identified novel ricin-type (R-type) lectin by sequencing of random clones from cDNA library of the coleopteran beetle, T.molitor. The cDNA sequence is comprised of 495 bp encoding a protein of 164 amino acid residues and shows 49% identity with galectin of Tribolium castaneum. Bioinformatics analysis shows that the amino acid residues from 35 to 162 belong to ricin-type β-trefoil structure. The transcript was significantly upregulated after early hours of injection with peptidoglycans derived from Gram (+) and Gram (-) bacteria, beta-1, 3 glucan from fungi and an intracellular pathogen, L. monocytogenes suggesting putative function in innate immunity.

This study was conducted to determine the effect of six different leguminous seeds a) cowpea, b) white kidney bean c) soybean (Cheongja) d) soybean (Daepung) e) mung bean and f) azuki bean on the nymphal development, mortality, adult longevity, fecundity and hatchability of Riptortus pedestris (F.) (Hemiptera: Alydidae). Total developmental time ranged from 18 to 25 days; R. pedestris fed on white kidney bean took the longest time. There was no difference in mortality among the bugs fed on the legumes except those fed on white kidney bean which had the highest mortality (94%). Also, adult bugs fed on white kidney bean showed the shortest longevity. Total number of eggs laid by adult female and the hatchability of the eggs were highest from the bugs fed on cowpea. Higher reproductive success of R. pedestris fed on cowpea suggests that cowpea could be very suitable legume for the bug and it can be used as a trap crop against the bean bug.

Several pentatomids and an alydid are reported inflicting losses in several host plants ranging from leguminous crops to fruit orchards in Korea. Understanding their visual preference would help develop mass trapping or efficient monitoring device. Visual preference of the major pentatomids and an alydid bugs to different colors and light emitting diodes (LED) of different wavelengths were observed. Adults of pentatomid such as Halyomorpha halys Stål, Plautia Stali Scott, and Nezara sp., and an alydid Riptortus pedestris (F.) were exposed to seven different colors (black, yellow, green, blue, white, cream, orange) in a multiple choice tests. Also, adult and second instar nymphs of the bugs were released inside a hexagonal LED boxes consisting of LED lamps of six different wavelengths (375 to 630 nm). Most of the pentatomids showed highest preference to blue color followed by yellow and 530 nm LED. However, R. pedestris showed higher preference to the yellow color followed by blue and black. Second instar nymphs of all the tested species also showed higher preference to 530 nm. Their color and LED lamp preference would help in development of mass trapping device.

본 논문은 1979년과 2006년 사이 전세계 강수의 연평균 및 계절성의 시계열적 변화 양상을 색채를 체계적으로 사용하여 공간상에 가시화하는 방법론을 제시한다. 시계열 푸리에분석을 이용하여 분해된 중간 결과물에 로버스트 및 비모수 시계열 회귀분석을 적용하여, 세계 전반적으로 강수량의 연평균 및 계절성이 어떻게 변화하는지를 GIS/RS 프로그램인 이디리시를 사용하여 지도화한다. 본 논문에서 제시한 지도학적 방법론은 세계 강수 변화의 시공간적 역동성을 하나의 지도상에 함축적으로 표현할 수 있으며, 각 지역에서 관찰되는 각기 다른 변화 양상을 2차원 색채 범례를 통해 명확히 구분한다. 또한, 이 방법론은 기후변화와 관련된 연구 외에도 다른 래스터기반 시공간 자료 표현을 하는데 있어서도 유용하게 활용될 것이다.