Jeju Island has been facing to threat of high-risk invasive pests from tropical areas. To protect domestic agriculture from those invasive pests, APQA has conducted a regular monitoring program on Jeju Island. We collected especially phototactic heteroceran pests by light bucket trap and identified them using their superficial appearances, and also mitochondria COI gene. As a result, a total of 24 families, 136 genera, 193 species, and 819 individuals were collected from around Jeju Island in 2023. Among them, two unidentified epidopteran pests, Palpita sp. (Crambidae) and Xyrosaris sp. (Yponomeutidae) were collected. In the present study, we report two unidentified micro-lepidopteran pests using superficial characteristics and mitochondria COI gene.

우리는 솔수염하늘소(Monochamus alternatus) 장에 존재하는 공생미생물들을 분리하였다. 그중 다양한 식물 들을 대상으로 성장 촉진 효과가 보고된 세균을 단일배양 분리하였다. 이 세균은 16S rRNA sequencing을 통하여 Enterobacter roggenkampii로 동정되었다. 우리는 분리된 E. roggenkampii에 대하여 genomic sequencing을 수행하 였고 유전학적 특성을 확인하였다. 우리는 E. roggenkampii가 식물의 성장을 촉진할 수 있는 다양한 유전자들을 가지고 있는 것을 확인하였고 그 중 IAA-Asp hydrolase 유전자를 가지고 있음을 알 수 있었다. 동시에, 분리된 E. roggenkampii와 같은 속의 세균을 대상으로 다양한 API kit와 기질 첨가 배지를 이용하여 생화학적 특성을 비교하였다. 향후 IAA-Asp 가수분해효소를 생산하는 잠재적인 식물 생장 촉진 비료 미생물로 등록하여 다양한 작물을 대상으로 생장 촉진 효과를 확인할 것이다.

Here we review the genus Nycteola of the Nolidae (Lepidoptera). As the result of this study, we recognized already synonymized species N. costalis Sugi, 1959 of N. coreana (Leech, 1900) and newly recorded species N. dufayi Sugi, 1982. Whlie the number of Korean Nycteola species remain unchanged, the composition has been changed.

우리는 길앞잡이(Cicindela chinensis)의 장에서 다양한 공생 미생물들을 분리하였다. 그중 다양한 곰팡이 성장 을 억제하는 세균을 동정하였고 “Ch-1”이라 명명하였다. 우리는 Ch-1 균주를 사용하여 10종의 식물 병원성 곰팡 이와 2종 곤충 병원성 곰팡이의 생장 억제를 확인하였다. 또한 8종의 항생제에 대한 저항성을 확인하였다. 동시 에, 본 균주의 genomic sequence를 수행하였고 유전적, 생화학적, 생리적 특성을 조사하였다. Ch-1균주는 특허등 록과 친환경 미생물제제로 등록하였고 향후 생물학적 방제제로써 활용될 수 있을 것으로 판단한다.

The baculovirus expression system (BES) utilize the p10 or polyhedrin promoter, a very late promoter that exhibits strong transcriptional activity primarily at the end of viral infection, to produce useful recombinant proteins. The burst sequence of the very late promoter is essential for strong transcription, and VLF-1 is a transcription factor that binds specifically to the burst sequence, and it has been reported that it can regulate the amount and timing of expression of protein by the very late promoter. Recently, a VLF-1 constitutively expressing cell line was constructed to increase the production of the target protein, but the effect was minimal. In this study, to find the optimal VLF-1 expression conditions to increase target protein production efficiency, we controlled the expression of VLF-1 through various promoters and evaluated the target protein expression efficiency by the p10 promoter accordingly.

Hand, foot, and mouth disease (HFMD) is a highly contagious disease with no specific treatment. Since it is common in immunocompromised children under the age of 5, there is a need to develop a safe vaccine. Virus-like particles (VLPs) are similar structures to viruses with the lack of genetic material which makes them impossible to replicate and infect, and therefore have a high level of biological safety and are considered to have high value as vaccines. In this study, the insect virus expression system that is widely used for vaccine and drug production due to its high post-translational modification efficiency, was used to produce VLPs for Coxsackievirus type A6 and A10, which are recently reported to be the main causes of HFMD. For this purpose, the selection of promoters that can control the timing and intensity of expression of 3CD protein, which is essential for VLPs assembly but has been reported to be cytotoxic, was conducted to construct an optimal expression form for HFMD-VLP.

Metarhizium은 대표적인 곤충병원성 진균 중 하나로, 종에 따라 매우 다양한 곤충에게 병원성을 일으키는 폭 넓은 기주범위를 형성한다. 이들이 주로 생성하는 것으로 알려진 destruxins (DTXs)이라는 2차 대사산물은 살충 활성 뿐만 아니라 항바이러스, 항증식, 항암 등 다양한 분야에서 효능이 연구되고 있어, 해당 물질에 대한 관심이 집중되고 있다. 살충 물질로서의 DTXs는 여러 곤충에 있어 병원성을 나타내는 것이 확인되었으나, 해충 으로서 전 세계적으로 심각한 경제적 피해를 일으킴과 동시에, 화학 살충제 저항성 문제가 야기되고 있는 목화진 딧물에 대해서는 아직 DTXs의 역할이 연구되지 않고 있는 실정이다. 본 연구에서는 목화진딧물에 대해 곤충병원 성 진균 Metarhizium spp.의 병원성 발현에 DTXs가 미치는 역할을 간접적으로 확인하기 위해, qPCR을 활용하여 진균 처리 후 목화진딧물 사망 시간과 관련하여 충체 내 DTX 합성효소의 발현을 비교 분석하였다.

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.

Aphis gossypii is a representative pest that transmits plant viral diseases. It is difficult to control with chemical pesticides alone due to their high pesticide resistance. Entomopathogenic fungi are biological control agents that can replace chemical pesticides and have characteristics of high host specificity and safety to humans. Therefore, we investigated the immune pathways of aphids against initial infection by entomopathogenic fungus. We treated aphids with the Beauveria bassiana JEF 544 strain and examined the immune response in early infection by qPCR. furthermore, we also studied changes the molting time of nymphs and changes in adult nymphal production caused by entomopathogenic fungi.

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.

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.

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.

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.