Mycoviruses are a group of viruses that infect filamentous fungi. While most hosts infected with mycoviruses do not show any symptoms. In some cases, mycoviruses induce various phenotypic changes include alterations in morphology, drug resistance, pathogenicity, virulence, sporulation, and growth. Entomopathogenic fungi are one of the integrated pest management agents as an alternative to conventional insecticides. Mycoviruses have the potential as supportive agents, enhancing the efficiency of the insecticidal activity of the fungi. Studies about mycoviruses themselves and their interaction with their hosts, especially entomopathogenic fungi, are needed to realize their full potential. In this work, the sequence of the dsRNA element isolated from the entomopathogenic fungus Metarhizium pinghaense 4-2 strain was determined. Through this study, we report the sequence of a dsRNA virus isolated from the Metarhizium pinghaense for the first time. In further studies, the ORF of the mycovirus that induces a phenotype change in the host will be researched.

마이코바이러스는 곰팡이를 감염시키는 바이러스로 자낭균류, 담자균류 및 난균류에서 주로 발견되며 일부 의 경우 곰팡이의 표현형에 영향을 끼치는 것으로 알려져 있다. 이번 연구에서는 대한민국 토양 샘플에서 분리된 65개의 자낭균류 및 접합균류 균주의 전체 핵산을 추출하고, 전기영동을 통해 바이러스 특이적 밴드를 스크리닝 하였다. 스크리닝 결과 65개의 균주 중에서 Tolypocladium spp. 균주 2개와 Marquandomyces marquandii 균주 1개에서 바이러스 특이적 밴드를 발견하였다. 그 후, Cellulose Chromatography를 이용하여 double-stranded RNA 를 분리하고 DNase I 및 S1 Nuclease 처리를 통해 DNA와 single-stranded RNA를 제거하여, Tolypocladium sp. 균주 1개와 Marquandomyces marquandii 균주에서 발견한 특이적 밴드가 dsRNA임을 확인하였다. 추후 virus-free isogenic line을 확보하여 virus 유무에 따른 표현형 변화를 확인하고, 마이코바이러스와 곰팡이 간의 상호작용에 관해 연구할 계획이다.

차세대 염기서열 분석(Next Generation Sequencing, NGS)은 대량의 병렬 데이터 생산으로 유전체의 염기서열 을 고속으로 분석하는 기술이며, 이 기술은 바이러스 유전체 분석에도 광범위하게 사용되고 있다. 하지만, 바이 러스의 전장 유전체가 100kb를 넘을 경우, 동일한 raw data라도 분석 방법 및 소프트웨어 그리고 매개변수 (parameter)에 따라 유전체의 크기와 구조가 다르게 결정된다. 따라서 유전체가 큰 바이러스 분석 시, 최적화된 NGS 분석 방법을 선택하는 것이 중요하다. 본 연구는 장수풍뎅이 누디바이러스(Oryctes rhinocerous nudivirus, 120kb) 유전체를 기반으로, 다양한 Assembly 소프트웨어(metaviralSPAdes, metaSPAdes, velvet, shovill, Geneious, megahit)를 사용하여, 최적화된 NGS 분석 방법을 고안하였다. Assembly 소프트웨어에 따라 바이러스 유전체 크기와 특징(Single Nucleotide Polymorphism, Insertion&Deletion, repetitive genomic variants)의 차이를 확인하였 다. Assembly 소프트웨어 간의 차이가 있는 염기서열은 Sanger sequencing을 통해 재확인하여, 참조 유전체 (reference sequence)를 구축하였다. 이 참조 유전체를 기반으로 가장 정확한 Assembly 소프트웨어와 parameter를 평가하였다. 본 연구는 분석 방법에 따라 달라지는 유전체의 특성을 이해하고, 바이러스 유전체를 정확하게 구축 하는 분석 파이프라인을 제공할 것으로 기대된다.

The fall armyworm, Spodoptera frugiperda, has developed extremely high levels of resistance to chlorantraniliprole and other classes of insecticides in the field. As microRNAs (miRNAs) play important roles in various biological processes through gene regulation. we examined the miRNA profile of S. frugiperda in response to Chlorantraniliprole, Indoxacarb and Thiamethoxam. Transcriptome analysis showed significant changes in the abundance of some miRNAs after treatment of S. frugiperda larvae with LC20 concentrations of three insecticides. A total of 197 miRNAs were systematically identified from S. frugiperda, and 16, 9, 2 miRNAs were differentially expressed after treatments of three insecticides. Importantly, three miRNAs were significantly downregulated and three were upregulated by RT-qPCR after treatment the LC50 of three insecticides with S. frugiperda larvae. Microinjection of agomirs of these six miRNAs into S. frugiperda larvae resulted in significant changes in mortality rates when exposed to three insecticides. Additionally, we also screened potential target genes for some of differentially expressed miRNAs, which may play important roles in insecticide resistance development. These findings provide valuable insights into the molecular mechanisms of insecticide resistance and underscore the potential of miRNAs as targets for the development of novel pest control strategies in S. frugiperda.

Urbanization is a driving force of global biodiversity changes, and species that successfully adapt to city environments can become pests with the assistance of human factors. Here we present the first genomic data of Plecia longiforceps, an invasive pest exhibiting intensive outbreaks in the Seoul Metropolitan Area of Korea. HiFi and Pore-C sequencing data were used to construct a highly continuous genome assembly with a total size of 707 Mb and 8 major pseudochromosomes. Gene annotation using transcriptome data and ab initio predictions revealed significant numbers of genes related to detoxification and heat tolerance. Comparison to the Bibio marci genome showed high levels of synteny with some regions of chromosomal rearrangement. Our data will serve as an essential resource for population and functional genomic studies on dispersal and outbreaks of P. longiforceps, and facilitate research on eco-evolutionary processes of dipterans in urbanizing habitats.

A new fumigant, carbonyl sulfide (COS), has potential for use as a replacement for methyl bromide, yet its mechanism of toxicity to insects remains poorly understood. In this study, transcriptome analysis was performed on Tribolium castaneum malpighian tubules and fat bodies, which are known to play an essential role in energy storage and utilization in insect species. In total, upon exposure to COS, 3,034 and 2,973 genes were differentially expressed in the T. castaneum malpighian tubules and fat body, respectively. These differentially expressed genes comprise a significant number of detoxification-related genes, including 105 P450s, 18 glutathione S-transferases (GSTs), 82 ABC transporters, 25 UDP-glucosyltransferases and 42 carboxylesterases and mitochondrial–related genes, including 9 complex Ⅰ genes, 2 complex Ⅱ genes, 1 complex Ⅲ gene, 9 complex IV genes, 8 complex V genes from both malpighian tubules and fat body tissues. Moreover, KEGG analysis demonstrated that the upregulated genes were enriched in xenobiotic metabolism by ABC transporters and drug metabolism by other enzymes. We also investigated the role of carbonic anhydrases (CAs) in toxicity of COS using dsRNA treatment in T. castaneum. These results show that CA genes have a key role in toxicity of the COS. Furthermore, the results of transcriptomic analysis provide new insights into the insecticidal mechanism of COS fumigation against T. castaneum and eventually contribute to the management of this important stored grain pests.

The habitat of Drosophila melanogaster is the environment of fruit decay/fermentation which emits high concentrations of chemicals. Our recent studies revealed that D. melanogaster has been evolutionarily adapted to its habitat through tolerance to chemicals and induction of antimicrobial peptides (AMPs) plays an important role for chemical tolerance. To determine the correlation between AMPs and the chemical tolerance pathway, we hypothesized that expression of AMPs is induced by tissue damages or ROS caused by chemical exposure and AMPs activate antioxidant enzymes, thereby inducing chemical tolerance in D. melanogaster. Therefore, in this study, we investigated the induction levels of genes associated with necrosis (EGR and BSK), apoptosis (Dronc, Dcp1, and Drice), antioxidant physiology (SOD1, SOD2, CAT, Trxr1, GstD2, and GstD5), and SAM metabolism (Gnmt and Foxo) in D. melanogaster exposed to three chemicals, 2-phenylethanol, ethanol, and acetic acid. As a result, above genes were induced in chemical-exposed fly, and this supports our hypothesis of chemical tolerance pathway in D. melanogaster.

Honey bee plays an important role in pollinating plants. Recently, however, declines in honey bee populations have been reported in many countries, and pesticides have been pointed out as one of the factors contributing to honey bee loss. To determine the effects of pesticides on honey bee behavior, we investigated the homing ability of honey bee exposed to four pesticides (acetamiprid, imidacloprid, fenitrothion, and carbaryl). In addition, the changes in expression levels of genes associated with ‘learning and memory’ (cGMP-dependent protein kinase foraging, Kruppel homolog 1, Adenlyate cyclase 3, Early growth response protein 1, Hormone receptor 38) were examined after pesticide treatment in forager bee. The four pesticides tested in this study generally reduced the homing ability of foragers. In the examination of gene expression, learning and memory-related genes were induced by the exposure to acetamiprid, imidacloprid, and carbaryl, whereas fenitrothion decreased the expression of these genes in honey bee. Although further studies are needed, this suggests that pesticides may have negative effects on honey bee behavior and behavior-related gene expression.

This study focused on the genomic analysis of Anopheles kleini and Anopheles pullus, both vectors of vivax malaria within the Anopheles Hyrcanus group. Using Illumina NovaSeq600 and Oxford Nanopore platforms, we identified 126 and 116 contigs, along with 40,420 and 32,749 genes from An. kleini and An. pullus, respectively. The assembled genome sizes were 282 Mb for An. kleini and 247 Mb for An. pullus, which are within a similar range to the sizes previously estimated by digital PCR (249 Mb and 226 Mb). We are currently also estimating the genome sizes of other Anopheles spp. and manually curating key genes determining vectorial capacity.

Scabies, caused by an infestation of the skin with the itch mite (Sarcoptes scabiei), is highly contagious and classified as a prevalent neglected tropical diseases. The current diagnostic approach relies solely on clinical judgment based on symptoms, history, and microscopic observation by an experienced dermatologist. To enhance sensitivity and specificity, we developed an alternative method based on mite-derived DNA. Our method involves a quick DNA release from skin scraping samples and Loop-Mediated Isothermal Amplification (LAMP) targeting the scabies mite-specific DNA sequences, enabling diagnosis within 30 minutes. Importantly, no cross-reactivity was observed when the sample was contaminated by two house dust mite species, and false positives were barely detected. Currently, we are in the process of developing a Point-of-Care Testing (POCT) kit for a scabies survey targeting school-age children in Timor-Leste as a global health project.

Parasites have co-evolved with their host for a long period of time, resulting in unique parasitic systems tailored to each host species. This makes them suitable for research on physiological function control through cross-species molecules like miRNA. The body louse, a vector of bacterial pathogens, is particularly valuable as a model insect due to their frequent feeding on human blood, which results in the continuous ingestion of human-derived miRNA and injection of salivary gland-derived miRNA into the human body. In this study, we conducted miRNA sequencing on body lice with mixed stages and identified 105 miRNAs, including 50 novel miRNAs. Sequence analysis of human miRNAs remaining in body lice and the functional analysis of these miRNAs are in progress.

The recent increase in the occurrence of common bed bug and tropical bed bug in shared areas highlights the need for rapid species identification at infestation sites, which is crucial for implementing targeted control measures due to differences in genetic and physiological traits. In this study, molecular diagnostic methods were developed using species-specific ITS2 sequences. Both multiplex PCR and loop-mediated isothermal amplification (LAMP) protocols with a DNA release method successfully distinguished between the two bed bug species regardless of developmental stages in 0.5~2.5 hours, even with dead specimens. Especially, LAMP's simplicity and speed make it applicable for rapid and accurate bed bug diagnosis at infestation sites.

천궁(Cnidium officinale Makino, COM)은 산형과(Umbelliferae)에 속하는 산림약용자원 중 하나로 뿌리나 지하 부를 건조한 후 약용으로 사용하는 여러해살이 산림약초 중 하나이다. 천궁(COM)은 고혈압, 진통 및 진정 등에 효과가 있다고 알려져 있어, 산림약용자원으로서 재배되는 품목 중 하나이지만 해충의 피해가 심각하여 방제에 어려움이 많이 있다. 천궁은 뿌리가 약재로 사용되지만 종근에 피해를 가하는 응애류 및 파리류 해충은 지하부에 활동하는 생태특성을 가지고 있어 피해초기에 발견하기 어려울 뿐만 아니라 방제가 어려운 실정이다. 천궁 (COM)은 연작이 되지 않은 작물 중에 하나이므로 재배포장을 매년 바꿔가면서 재배되고 있는 실정이다. 수확시 기는 가을이므로 재배하는 동안에는 정확한 병해충조사가 쉽지 않을 뿐만 아니라 정확한 발생시기 및 피해정도 를 확인하는 것이 어려운 실정이다. 본 연구의 목적은 천궁의 해충 및 천적곤충에 대해 조사하여 천궁의 해충종류 및 발생에 대해 이해하고 이를 기반으로 친환경 방제를 위한 천적류의 정보수집 및 분석 등을 위한 기초자료로 제공하고자 한다.

Spodoptera exigua is one of the worldwide distributed agricultural pest insects and has been known to show high resistance to conventional chemical insecticides. Autographa california multiple nucleopolyhedrovirus (AcMNPV) has been used as eco-friendly biological control agent for S. exigua, as it exhibits high level of host specificity, stability and safety. In this study, for formulation of AcMNPV, the optimal conditions for mass-production of AcMNPV polyhedra was established using S. exigua larvae. Mass-produced AcMNPV polyhedra was formulated as wettable powder using microencapsulation method and its control efficacy against S. exigua was evaluated both in laboratory and semi-field experiment. Chinese cabbage treated with the AcMNPV formulation showed significantly reduced damage rates, suggesting that the AcMNPV formulation in this study could be useful for control of S. exigua

Bacillus thuringiensis (Bt) is widely used as an environmentally friendly insecticide compared to chemical insecticides. However, challenges such as difficulty in direct practical application, limited efficacy duration, and stability have been identified. To solve these issues, formulation-based research is being extensively conducted. In this study, the high insecticidal activity strain Bt IMBL-B9, identified in previous research, was subjected to large-scale cultivation using a fermentor. Subsequently, various formulations were developed through formulation processes. and characteristics such as their wettability, suspensibility and particle size were assessed to select the optimized formulation.

The two-spotted spider mite, Tetranychus urticae Koch, is one of the economically important agricultural pests globally, as it attacks a wide range of vegetable and horticultural crops. In this study, we evaluated spatial repellent and oviposition deterrent activities of T.urticae in response to fifteen compounds derived from ester-containing natural products. To evaluate the tests, we used bridge two-choice test and host two-choice test in laboratory conditions. Among the eight compounds showed spatial repellent and oviposition deterrent activities against T. urticae at the 20 mg dose and some compounds had the activities at lower dose. We also conducted two-choice test with a blend and single compounds to determine which showed stronger spatial repellent and oviposition deterrent activities. In host two-choice test, we evaluate repellence between distance of compounds. This study concluded that series compounds from ester-containing natural products have the potential to be used managing T. urticae in the field.

Entomopathogenic fungi serve as eco-friendly alternatives to chemical pesticides. In this study, we investigate the interactions between mosquitoes and Metarhizium anisopliae JEF-157, which showed high insecticidal activity against mosquitoes, by RNA-seq analysis. RNA from mosquitoes was extracted at the median lethal time to identify changes in gene expression. The results showed 580 genes were up-regulated, while 336 genes were down-regulated in fungal treated mosquitoes. Up-regulated genes were related to metabolic and cellular processes such as cytochrome P450, DNA replication, and apoptosis. Down-regulated genes were involved in metabolism pathways such as lysosome, starch and sucrose metabolism, and fatty acid biosynthesis. These results are crucial for elucidating the mechanisms of fungal invasion and interaction in insects, providing insights for future pest management strategies.

Aphids are well-known insect pests that, due to their feeding habits, affect various crops' productivity and marketability. Additionally, they cause significant damage worldwide as vectors of viruses. Chemical pesticides are widely used to control these agricultural pests. However, due to resistance to various chemical pesticides, pest control is becoming difficult in agricultural environments. Entomopathogenic fungi such as the genus Metarhizium and Beauveria are being extensively researched as alternatives to these chemical pesticides. In this study, the mycological properties of isolated from soil Metarhizium pemphigi IPBL-H and the insecticidal activity of this strain against Myzus persicae and Aphis gossypii were evaluated for potential of development as a biopesticide.

Entomopathogenic fungi have been studied to control insect pests as an alternative to chemical insecticides. However, all fungi haven't a high virulence against pests. In this study, we compared the biological characteristics of Metarhizium anisopliae strains. First, we selected four M. anisopliae strains and compared the thermotolerance, conidial productivity, and virulence. For the thermotolerance test, conidial suspensions were exposed to 0, 30, 60, and 90 min at 45 °C. As a result, the conidial germination rates were over 95% when exposed for 0 min but, were 64, 37.7, 6, and 3% when exposed for 30 min at 45°C, respectively. To compare conidial productivity, 200g of millet were used and inoculated with a conidial suspension of 1 ml (1×107 conidia/ml). Conidial productivity was investigated after 14 days. As a result of conducting a virulence test against mealworms using a spray method, differences in virulence between strains were confirmed.

본 조사는 2023년 국가생물다양성 기관연합 공동조사의 일환으로 제주도 곶자왈도립공원 및 선흘곶 동백동 산 일대의 곤충류 분포현황을 파악하고, 자연자원의 발굴 및 관리에 대한 기초자료를 확보하고자 수행되었다. 곤충류 현지조사는 주요 곤충류가 출현하는 시기인 2023년 5월 22~26일 및 8월 28일~9월 1일 총 2회에 걸쳐 채집 조사가 수행되었다. 조사방법으로는 조사지 주변에 깔때기 트랩을 설치하여 조사 마지막날 수거하였으며, 주간 에는 포충망을 이용하여 채어잡기, 쓸어잡기, 털어잡기, 직접찾기 및 사진촬영 등을 수행하였다. 야간에는 조사 시 식생이 양호한 지역을 선정하여 일몰 전에 유아등을 설치한 후 자정 이전까지 야간조사도 병행하였다. 현지조 사를 통하여 채집된 곤충은 연구실로 가져온 후 건조표본 및 액침표본으로 제작되었으며, 각 분류군별 도감을 참고하여 종 수준까지 분류 및 동정 작업을 수행하였다. 조사결과 총 10목 69과 264종의 곤충류가 조사되어 비교 적 다양한 분류군이 출현하는 것으로 확인되었다. 또한 금번조사 결과 멸종위기야생생물 Ⅱ급에 해당되는 애기 뿔소똥구리 1종이 확인되었으며, 국외반출 승인대상종 46종, 유용곤충 17종, 분포특이종 12종, 해충 9종, 고유종 5종 등을 포함하여 총 70종의 특정종이 확인되었다.