국립수목원 산림생물다양성연구과는 2023년부터 산림생태계 보전을 위한 화분매개곤충 특성연구 사업을 추진하면서 그 첫 번째 단계로 화분매개곤충 인벤토리 구축을 위한 연구를 수행하고 있다. 수목원 4개소(강원도 립화목원, 경상남도수목원, 국립수목원, 대구수목원)에서 9종의 식물(국수나무, 미선나무, 산수유, 산철쭉, 생강 나무, 수수꽃다리, 아까시나무, 진달래, 히어리)을 대상으로 각 식물에 어떤 곤충들이 방문하는지 쓸어잡기 방법 을 이용하여 확인하였다. 조사된 곤충 중에서 가장 많은 개체수를 차지하고 있는 곤충은 파리목(1,407개체)이었 으며, 26과의 다양한 파리목 곤충들을 확인할 수 있었다. 그중에서도 가장 많은 개체수를 차지하고 있는 과는 재니등에과 (4속, 5종, 651개체), 꽃등에과 (24속, 34종, 302개체), 기생파리과 (8속, 9종, 111개체)로 집계되었으며 이 중에서도 꽃등에과의 종다양성이 가장 높게 나타났다. 조사한 식물 중에서 가장 많은 파리목 곤충이 모인 식물은 수수꽃다리와 미선나무였고, 각각 314, 308개체 이상의 파리목 곤충을 확인하였다. 반면 파리목 곤충의 방문이 가장 저조한 식물은 국수나무와 아까시나무였으며 확인된 개체수는 각각 38, 45개체였다. 우리는 화분매 개곤충에 대한 인벤토리 구축을 통해 화분매개곤충에 대한 다양한 분류학 및 생태학적 데이터의 지속적인 확보 를 위해 노력하고 있으며, 본 소모임에서 이에 대한 발전방안 및 축적된 데이터의 활용방안에 대해 모색하고자 한다.
Since the gene expression interference induced by dsRNA was discovered, dsRNA has been considered as an antiviral agent and pesticide to protect beneficial insects and crops, respectively. Recently, dsRNA was classified as IRAC mode of action group 35, and the first dsRNA pesticide, Calatha of GreenLight Bioscience, has been approved by EPA. Also an animal drug for Asian honeybee, HoneyGuard-R of Genolution is about to be approved by APQA. During the last two decades, hundreds of papers already had demonstrated the application and capability of dsRNA for agriculture, however, we have just a few commercialized products at hand at this moment. It is time to understand the processes, hurdles and limitations on the industry side that are indispensable for the development, registration and commercialization of dsRNA-based products.
RNAi (RNA interference, RNA 간섭)은 타겟 유전자의 mRNA 시퀀스와 상동한 이중 가닥 RNA (double-stranded RNA)에 의해 일어난다는 사실이 밝혀진 이후, 유전자의 기능을 파악하는 연구 도구로 활용되다가, 2018년에 비로소 사람의 유전자 치료제로 상용화가 되었다. 지금은 더 많은 질병들의 유전자 치료제를 만들기 위해 다양한 회사들이 임상을 진행하고 있다. 농업의 분야에서는 곤충의 ‘생존 필수 유전자’를 타겟함으로써 해충 방제제로 개발되어 서부 옥수수 뿌리벌레(Western corn rootworm)를 RNAi로 방제하기 위한 제품이 종자 형식으로 상용화 되었고, 최근에는 심각한 농약저항성을 가진 콜로라도 감자벌레(Colorado potato beetle)를 RNAi로 방제하기 위 해 ‘스프레이 방식의 뿌리는 dsRNA’가 상용화되었다. 추후에 더 많은 해충을 타겟으로 하는 제품이 개발될 예정 이다. 본 발표에서는 위에 언급한 상용화된 dsRNA 시퀀스의 환경안전성평가와 관련된 연구, off-target effect, siRNA 활용, nanoparticle 등 RNAi 기술을 보다 안전하고 유용하게 농업적으로 사용할 수 있는 방법에 대한 ‘실험 실의 역할’이 무엇인지 알아보고자 한다.
Protaetia brevitarsis larvae have been widely used for traditional medicine and food in East Asia. This research comprised two experiments. The first experiment compared the growth in densities (10, 20, 30 larvae), substrates (commercial, self-made), and containers (Tyvek, zipper bags). In the second experiment, different numbers of air holes (16, 32, 48 holes) in zipper bags experiment is ongoing. The results indicated that the larvae development rate was 72.6%, 69.98%, and 55.33% in 10, 20, and 30 larvae densities, respectively. However, there was no significant difference in larvae survival rate and weight. In the different feed material experiments, the average larvae weight of commercial feed was 1.09g and for self-made, it was 2.85g. The survival rate was 86% and 96% in commercial and self-made feed, respectively. Lastly, while 96% of larvae survived in Tyvek bags, none of the larvae survived in the zipper bags.
Stoneflies (Plecoptera) are known for being sensitive to water pollution and are used as bioindicators for evaluating water quality. Among them, Nemouridae, especially the genus Nemoura, which are commonly referred to as winter stoneflies, can be found around streams even during the cold winter months. Nemoura geei Wu, 1929, among them, was originally described from Beijing and is widely distributed in Korea, China, Japan, and the Russian Far East. Here, we report the development and characterization of new functional microsatellite markers of N. geei using high-throughput sequencing technology. A total of 80,661 microsatellite loci were identified with a total length of 1,801,591 bp. The average length was 22.34 bp, and microsatellites occupied 0.42% of the entire sequence. The novel 20 microsatellite markers developed in this study can be usefully applied to the population genetics analyses as important genetic resources for understanding the ecological and evolutionary characteristics of a stonefly species at the population level in Korea.
상추는 수요가 연중 지속됨에 따라 재배면적과 생산량이 증가하고 있는데, 주로 생식으로 이용되며 수확간격 이 짧아 해충 다발생시 약제방제가 어려운 작물이다. 최근 검은무늬밤나방이 약제방제가 소홀한 포장에서 잎을 광포식하여 빠른 시간내에 직접적인 피해를 주고 있다. 본 연구에서는 검은무늬밤나방의 온도별 발육특성을 구명하여 방제를 위한 기초자료로 이용하고자 한다. 검은무늬밤나방은 시설재배 상추에 발생한 유충을 채집하 여 실내 사육 후 항온조건(15, 20, 25, 30℃)에서 발육단계별 발육기간을 조사하였다. 온도가 높아짐에 따라 각 태별 발육기간은 짧아지는 경향이었으며, 유충은 보통 4회 탈피하였다. 25℃에서 알기간은 2.5일, 유충기간 10.0 일, 전용기간 1.0일, 번데기기간 7.0일로 알에서 성충까지의 발육기간은 20.5일 이었다.
With the increasing indroduction and spread of invasive quarantine pests, accurate diagnosis of pests detected in quarantine sites has become crucial. DNA barcoding, a standardized method that complements morphological analysis for rapid and precise species identification, is actively researched worldwide. In this study, we established a molecular biological identification system for major pests encountered during the import and export of agricultural and forestry products. By analyzing the DNA barcode sequences of pests collected domestically and those detected in quarantine inspections, we compiled genetic information for 1,292 individuals representing 472 species, 108 families across 11 insect orders. Among these, order Lepidoptera had the highest diversity, with 251 species across 27 families. We also secured barcodes for 52 species, 24 families in order Hemiptera, and 70 species, 20 families in order Coleoptera. By constructing a comprehensive biological foundation and database for various pests detected in quarantine sites, we aim to enhance the quarantine system by enabling rapid and accurate identification of invasive pests, thereby blocking early.
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.
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.
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
Six mosquito species in the Anopheles Hyrcanus group are known as vectors responsible for transmitting vivax malaria in South Korea. In this study, seasonal dynamics of Anopheles Hyrcanus group species and knockdown resistance (kdr) mutations in malaria-endemic sites, Paju and Gimpo, were monitored over two years. In August 2023, all six species were observed simultaneously in one of the Paju collecting sites, and kdr mutations were newly identified in all species except Anopheles kleini. Although Anopheles pullus was revealed as a relatively resistant species among five species populations without kdr mutation via bioassays, there were no critical differences in the voltage-sensitive sodium channel sequence. These findings suggest variability in pyrethroid resistance mechanisms among Anopheles Hyrcanus Group species.
RNA interference (RNAi) has been applied to control insect pests using gene silencing machinery in which small interfering RNA derived from dsRNA specifically degrades target mRNA. This study optimized dsRNA insecticide specific to thrips infecting hot peppers. Among potent candidate target genes, vATPase B was chosen because its RNAi was highly efficient as much as Snf7, a well-known RNAi target gene. Although RNAi specific vATPase B is lethal to Frankliniella occidentalis, it was not much effective to control other thrips species such as F. intonsa and Thrips tabaci. To expand its target spectrum, we devised a mixture treatment of dsRNA specific to individual species. As expected, each dsRNA was highly efficient in a species-specific manner. This supported the hypothesis of 21mer identity for the efficient RNAi. However, the dsRNA mixture efficiently killed the three thrips species in a crop field. To further expand its spectrum to the whitefly, Bemisia tabaci, we applied virus-induced gene silencing (VIGS) to produce dsRNA in the hot peppers using Tobacco Rattle Virus. VIGS successfully suppressed control gene. dsRNA produced by VIGS gave significnat mortality to B. tabaci in addition to the thrips. These results suggest a technique to expand dsRNA insecticide spectrum using a mixture treatment and VIGS in insect pest control_.
To overcome the challenges in tracking insects underground, a detection method consisting of a metal detector and aluminum tag was developed for tracking a dung beetle, Copris ochus (Coleoptera: Scarabaeidae). First, detection rate was evaluated for varying volumes of aluminum tags varying orientations of the tags under soil. Then, the detection efficacy was evaluated in the field at varying depths of hidden tags in two types of vegetation. Finally, the effect of aluminum-tagging on the survivorship, burrowing depth, and horizontal movement of C. ochus adults were assessed. Generally, an increase in tag volume resulted in a greater detection depth with maximum depth of 17 cm. Orientation, however, did not affect detection rate except when tag was placed perpendicular to the soil surface. In the field, metal detectors could detect aluminum-tagged models with success rates ≥85% up to 10 cm and 45−60% at 20 cm under soil. Finally, no significant effect of tagging on survivorship and behaviors of C. ochus was observed.
Spodoptera eridania and S. ornithogalli (Lepidoptera: Noctuidae), which are polyphagous pests that damage various crops such as tomatoes and beans are regulated quarantine species that are highly likely to invade South Korea. Therefore, it is crucial to promptly and accurately identify the presence of S. eridania and S. ornithogalli in crop fields to effectively eradicate as a regulated quarantine species. In this study, we developed a loop-mediated isothermal amplification (LAMP) assay, which allows for rapid in-field identification. To develop the LAMP assay, we selected target species-specific genomic regions from the whole-genome sequences of one target and 13 other lepidopteran species. We validated each five and six primer sets that consistently produced positive reactions in S. eridania and S. ornithogalli, respectively. To test the sensitivity of the each locus, LAMP reactions were performed using various reaction times using crude DNA, which was extracted from various types of adult tissues. All sensitivity tests were also successful.
Density survey should be carried out for applying integrated pest management strategies, but it is labor-intensive, time- and cost-consuming. Therefore, binomial sampling plans are developed for estimating and classifying the population density of whiteflies late larvae based on the relationship between the mean density per sample unit (7 leaflets) and the proportion of leaflets infested with less than T whiteflies ( ). In this study, models were examined using tally thresholds ranging from 1 to 5 late larvae per 7 leaflets. Regardless of tally thresholds, increasing the sample size had little effect on the precision of the binomial sampling plan. Based on the precision of the model, T=3 was the best tally threshold for estimating the densities of late larvae. Models developed using T=3 validated by Resampling Validation for Sampling Plan program. Above all, the binomial model with T=3 performed well in estimating the mean density of whiteflies in greenhouse tomato.
The brown planthopper (BPH) and white-backed planthopper (WBPH), significant rice pests, cannot overwinter in Korea and primarily originate and migrate from Southeast Asia and China, where they cause considerable damage. These planthoppers, along with other pests, annually migrate to the Korean Peninsula. Monitoring their migration is essential for controlling their populations and maintaining agricultural productivity. Traditional monitoring methods often struggle with timeliness due to time and manpower constraints. To address this, we developed the Smart Aerial Net Traps (SANT) for immediate tracking of pests. The SANT system is installed in 43 locations across the country and has been used for over 10 years to track migrating insects. Our research shows that SANT is a more effective method for monitoring migratory pests compared to traditional methods. SANT enables real-time tracking of various migratory pests and can also be utilized in different areas, such as analyzing pest population changes and determining pest origins through the study of air currents.
교량은 국가의 사회경제 발전과 국민 안전에 중대한 영향을 미치는 중요한 인프라이다. 하지만 다수 의 국내 교량이 1970년대와 1980년대의 고도의 경제성장기에 완공되었으며, 현재 교량의 노후화 비중 은 전체 교량의 30%에 달하고 있다. 현재 노후화 교량에 대한 유지 보수 시스템의 부족으로 인해 교 량 붕괴 사고가 발생한 사례들이 발생하고 있다. 따라서 노후화가 진행되는 교량에 대해 효율적으로 관리하기 위한 우선순위 평가 모델 개발은 중요시된다. 본 연구에서는 개발한 유지관리 우선순위 모델 을 개발하였고, 이를 실제 교량에 대해 적용하여 평가하였다. 제안된 모델의 활용을 통해 위험에 노출 된 교량의 유지 보수 및 관리를 효율적이고 신뢰할 수 있게 수행할 수 있을 것으로 판단된다.
본 연구에서는 바이오차 콘크리트의 개발과 역학적 특성에 대한 연구를 통해, 환경친화적이고 지속 가능한 건설 자재로의 활용 가능성을 탐구하였다. 바이오차는 바이오매스와 숯의 합성어로, 탄소격리 효과가 있어 시멘트 대체재로 사용할 수 있는지 실험을 통해 분석하였다. 바이오차를 시멘트 질량비 5%를 치환 최대치로 설정하였다. 실험 변수로 바이오차의 시멘트 대체율을 0%에서 5%까지 1%씩 올 려 대체하여 방식으로 설정하였다. KS F 2403에 따라 시편을 제작하였고, 슬럼프, 압축강도, 쪼갬인장 강도, 휨강도를 실험을 통해 바이오차 콘크리트의 역학적 특성을 분석·비교하였다. 실험 결과, 바이오 차의 대체율이 증가할수록 슬럼프가 감소하는 경향이 나타났다. 압축강도는 바이오차가 시멘트를 대체 함으로써 강도가 감소하였지만, 대체율 1%(23.37MPa)를 제외한 실험체에서 설계기준 압축강도 24MPa 이상을 만족하였다. 휨강도는 대체율 5%가 가장 높았으며 0% 대비 약 12% 증가하였다.