수염풍뎅이(Polyphylla laticollis manchurica)는 과거에는 흔히 발견되었으나, 1970년대 이후 한반도 내 개체수 가 급격히 감소하여 2005년 환경부에 의해 멸종위기 야생생물 Ⅰ급으로 지정되었다. 또한 해당종의 분자생물학적 연구는 멸종위기종이라는 특성으로 인해 제한적으로 진행되었다. 그로 인해 NCBI 등 공공 데이터베이스에서 제공되는 서열정보들 또한 부족한 실정이다. 이 연구는 이러한 한계를 극복하고 수염풍뎅이의 유전적 특성을 규명하기 위해 생물정보학적 기술을 활용하여 전사체 분석을 진행하였다. Illumina HiSeq 2500 플랫폼을 사용하여 53,433,048개의 RNA reads를 얻었으며, Trinity와 TGICL을 이용한 De novo 어셈블리 분석을 통해 18,172개의 unigenes를 생성하였다. 생성된 unigenes는 GO, KOG, KEGG, PANM DB를 활용하여 annotation을 진행하였다. 그 결과, GO 분석에서는 ‘binding and catalytic activities’와 관련된 항목이 높은 발현을 보였으며, KOG 분석의 경우 ‘Cellular Processes and Signals’ 범주가 높은 비율을 나타내었다. KEGG 분석을 통해 2,118개의 unigenes가 metabolic 카테고리에 annotation된 것을 확인하였다. SSR 모티프 분석에서는 AT/AT (42.90%) 모티프, AAT/ATT (13.13%) 모티프 순으로 많이 나타나는 것을 확인하였다. 이 연구를 통해 분석한 결과 들을 이용하여 유전자원 및 종 정보를 실시간 제공 및 정보 공유가 가능하도록 Database 및 web-interface를 구축하 였으며, 이러한 자료들은 국내 멸종위기종인 수염풍뎅이의 고유한 유전적 특성을 발굴 및 확보할 수 있는 기반자 료로써 활용될 수 있을 것으로 사료된다.

장내 미생물 군집은 소화 과정, 면역 시스템, 질병 발생 등 숙주의 다양한 면에 광범위한 영향을 주는 것으로 알려져 있으며, 주요 장내 미생물 종은 숙주의 생리 기능에 핵심적인 역할을 수행한다고 발표된 바 있다. 곤충의 장내 미생물 군집에 관한 연구가 최근 활발히 이루어지고 있으며, 이들 연구는 주로 장내 미생물 군집과 기생충, 병원체 간의 상호작용, 종간의 신호 전달 네트워크, 먹이의 소화 과정 등을 중심으로 이루어지고 있다. 이러한 연구들은 대부분 Illumina MiSeq을 활용하여 16S rRNA 유전자의 V1부터 V9 영역 중 선택된 특정 부분을 대상으로 짧은 서열 정보를 대상으로 진행되었다. 그러나, 최근에는 PacBio HiFi 기술이 상용화되면서 16S rRNA의 전장 분석이 가능할 수 있게 되었다. 이번 연구는 장수말벌(Vespa mandarinia)의 해부를 통해 gut과 carcass 부분을 분리한 뒤, 각 샘플을 Illumina MiSeq과 PacBio HiFi 기술을 활용하여 미생물 군집 간의 차이점을 확인하기 위하여 수행되었다.

The blackish cicada, Cryptotympana atrata Fabricius, 1775 (Hemiptera: Cicadidae) was originally distributed mainly in the southernmost remote island, Jeju and rarely throughout low lands in South Korea, but has been explosively increased at the urban areas, where annual temperature is higher. In this study, we sequenced a partial mitochondrial COI from a total of 171 individuals collected throughout 12 localities in South Korea. The haplotype found with the highest frequency in Jeju island shares only with two inland localities in southern region with a low frequency, whereas the haplotype found with the highest frequency throughout inland localities was not found in Jeju island. These results showed that Jeju population, southern region, and other inland populations form somewhat different genetic groups.

The large copper butterfly Lycaena dispar (Haworth, 1803; Lepidoptera: Lycaenidae) has been categorized as a near-threatened species (NT) in South Korea from 2012 mainly due to limited distribution. In this study, we visited 36 sites spread across all South Korean provinces to verify the distributional range of the species and sequenced mitochondrial COI for 53 individuals from nine sites. We observed L. dispar at 15 sites in six provinces, including the two previously known provinces, indicating a southward range expansion. The in-field monitoring and genetic data collectively suggested that L. dispar does not have a limited distribution nor is it isolated, indicating that it should be reclassified as less vulnerable. Our study demonstrates that the combination of field and genetic data can provide a more reliable assessment of the stability of a species.

Bombyx mandarina (Lepidoptera: Bombycidae), the presumed ancestor of the domesticated silkworm B. mori, has long been a subject of study to illustrate the geographic relationships in connection with origin of B. mori. We sequenced 100 mitochondrial genomes of B. mandarina collected from South Korea and Japan and these were combined with public data. Phylogenetic and population genetic analyses showed that all individuals collected in South Korea, except one individual from Inje, formed a strong group together with northern China and some individuals of southern China. This group was placed as the sister group to B. mori strains, suggesting that this group could have been served as an immediate progenitor for B. mori. For further robust inference more analysis is underway.

단백질의 구조 예측은 생명 과학 및 의약학 분야의 핵심적인 연구 주제 중 하나로, 단백질의 기능 및 상호작용을 이해하기 위한 주요 정보를 제공할 수 있어 다양한 연구가 수행되고 있다. 이러한 연구의 일환으로 최근 Google DeepMind의 AlphaFold2가 등장하였으며, 단백질 구조 예측 성능을 대폭 향상시켜 CASP(Critical Assessment of Protein Structure Prediction)에서 뛰어난 평가점수를 받아 단백질 구조 예측 분야의 최신 기술을 크게 향상시켰다. 이러한 컴퓨터 기반의 단백질의 구조 예측 방법은, 고전적인 방법을 사용하여 직접 단백질 구조를 결정하는 방법 에 비해 매우 정확하고 빠르며 경제적인 비용으로 수행될 수 있어 단백질 구조 예측 및 생리학 연구를 수행하는 연구자들에게 유용한 방법론이 될 것으로 사료된다. 따라서 본 연구소에서는 곤충을 포함한 무척추 자생동물을 연구하는 연구자들을 위해 단백질 구조 예측을 수행할 수 있도록 64Core/128Threads의 CPU, 256GB의 RAM과 6장의 GeForce RTX 3090으로 이루어진 GPU(Graphical Processing Unit) 고성능 컴퓨터 시스템에 AlphaFold2 program을 구축하였다. 최근 인간을 대상으로 한 단백질 구조 예측 연구는 상당한 진전을 보이고 있지만, 곤충을 포함한 자연계의 동물을 대상으로 한 연구는 여전히 미비한 상황이다. 이러한 자생동물자원연구의 확대를 위해 본 연구소에서 구축한 GPU 시스템 및 생물정보학적 분석 방법이 많이 활용되어야 하며, 이를 위해서는 연구자들 의 협력과 참여가 필요하다.

Bombyx mandarina (Lepidoptera: Bombycidae), the presumed ancestor of B. mori, has long been a subject of study to illustrate the geographic relationships in connection with origin of B. mori. We report 97 mitochondrial genome (mitogenome) sequences of B. mandarina collected from Korea and Japan. Phylogenetic and population genetic analyses showed that all individuals of B. mandarina collected in Korean localities formed a strong group together with all individuals originated from northern China (mainly north of the Qinling-Huaihe line) and some of southern China. This group was placed as the sister group to B. mori strians suggesting that this group had been served as an immediate progenitor for B. mori.

Bumblebee, Bombus ardens ardens (Apidae: Hymenopera), is an important resource for pollination that is most widely distributed in Korea. This study utilized microsatellite markers for investigation of genetic diversity and geographic relevance of the B. a. ardens populations in Korea. Through Next Generation Sequencing analysis, we identified 10 microsatellite markers and genotyped for 107 individuals of B. ardens collected from 10 populations. At each locus the number of alleles ranged from 10 to 23; the observed and expected heterozygosities ranged from 0.8909 to 1.0000 and 0.6641 to 0.8422, respectively; and inbreeding coefficient(FIS) ranged from –0.5053 to –0.0891. Significant deviation from the Hardy–Weinberg equilibrium was not observed at any locus. Population structure analysis indicated that there are three genetic groups in Korea with each Jeongseon and Ulleung-do composed of different gene pool from the remaining other populations. Similarly, Principal coordinates analysis also showed the same pattern. FST and RST analyses showed that each Jeongseon and Ulleung-do population had a significant genetic distance from other populations. Considering these results, genetic isolation of Ulleung-do may be explained by “Oceanic island” status and Jeongseon, which showed the positive FIS (0.069) and genetic isolation may be caused by its location on the east side of Baekdudaegan and by on-going inbreeding with a small population size.

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.

Lepidoptera is one of the largest insect orders, but the phylogenetic relationships within this order, have yet to be completely described. One of the unresolved relationships includes the monophyly of Papilionoidea in relationship with the monotypic superfamily Hesperioidea. We newly sequenced five hesperid mitochondrial genomes (mitogenomes), representing four subfamilies: Pyrginae (Daimio tethys and Lobocla bifasciatus), Coeliadinae (Choaspes benjaminii), and Hesperiinae (Potanthus flavus), and Heteropterinae (Carterocephalus silvicola). Along with these newly sequenced hesperid genomes phylogenetic analysis was conducted with all available lepidopteran mitogenomes including three reported species of Hesperiidae that consisted of ~70 species in ten lepidopteran superfamilies. The test for the effect of optimization schemes, such as exclusion and inclusion of third codon position of 13 PCGs, other genes (22 tRNAs and two rRNAs), and with and without partitions also was performed. Majority of datasets consistently placed the monophyletic Hesperiidae the sister to ((Pieridae + Lycaenidae) + Nymphalidae), placing another true butterfly family Papilionidae as the basal lineage of this group, presenting the relationships (Papilionidae + (Hesperiidae + ((Pieridae + Lycaenidae) + Nymphalidae))). Consistent to previous result, Pyraloidea was placed as the sister to ((Bombycoidea + Geometroidea) + Noctuoidea), placing the Macrolepidoptera as non-monophyletic group.