Melon thrips, Thrips palmi Karny (Thysanoptera: Thripidae) is a worldwide polyphagous pest. The management of this pest have mainly relied on chemical agents. However, the overuse is harmful to the environment and results in insects resistance. Therefore, alternative eco-friendly control methods that have different mode of action, such as biological control, are necessary to overcome the current issue. In this study, we isolated entomopathogenic fungi from Korean soil, and characterized them via morphological and molecular techniques and pathogenicity assay against Tenebrio molitor larvae. The isolated fungi were screened for virulence against T. palmi under laboratory conditions and the results were used to establish a thrips-pathogenic fungal library. The highly virulent isolates were selected and further characterized for optimum culture conditions and application as biopesticide in the field.

수노랑나비(Chitoria ulupi (Doherty, 1889))는 한반도 전역에 국지적으로 분포하는 종으로, 국외의 경우 인도, 미얀마, 라오스, 중국, 대만 등에 서식하는 것으로 알려져 있다. 이 종에 대한 생활사는 김과 서(2012)에 의해 확인된 유생기와 번데기의 형태 등이 일부 보고 된 것 이외에는 알려진 바가 없다. 본 연구에서는 한국산 수노랑나비의 생태적 습성(성충 산란행동, 점유행동, 짝짓기, 먹이습성 등)과 미성숙단계 (알, 유충, 번데기)의 형태적 특징 등 생활사 전반에 대하여 보고 하고자 한다.

최근 기후변화와 더불어 사과원 병해충 방제 횟수, 주변 식생 등의 변화로 인해 노린재류의 발생 밀도가 증가하고 피해가 점점 늘어나고 있다. 사과원 노린재류의 방제는 주로 살충제를 이용하고 있으나 이들의 강한 이동성과 농약 잔류, 작업자에 대한 농약 노출 등의 문제를 야기하는 단점이 있어 이러한 문제를 해결할 수 있는 새로운 해충 방제 수단이 필요한 실정이다. 본 연구는 사과원의 주요 노린재 중 썩덩나무노린재와 갈색날개노린재의 알을 이용하여 사과원 주변의 알 기생봉을 탐색하고 이들에 대한 야외 기생률을 알아보고자 실시하였다. 저온 저장된 두 종의 알을 사과원에 배치하고 이후 수거하여 알에서 채집된 기생봉을 동정한 결과 썩덩큰검정알벌(Trissolcus japonicus Ashmead)로 확인되었다. 야외 기생률은 썩덩나무노린재의 경우 약 27%, 갈색날개노린재는 21%로 나타났다. 추후 이 기생봉에 대한 생태 특성, 인공 사육, 야외 방사 기술 등의 생물적 방제제로 활용하기 위한 다양한 연구를 진행할 예정이다.

A species of Beauveria bassiana is widely used for biological pest management in many countries. Many efforts have been given to figure out the clear fungal mode of action to enhance the insecticidal activity. Homologous recombination (knock-out) or hairpin RNA (knock-down) is popularly used in fungal gene function study, but gene cloning and generation of knock-out or -down mutants takes long time or temporarily knock-downed. Here in this work, we used previously generated egfp-expressing B. bassiana strain (Bb-egfp #3) and integrated dsegfp to the Bb-egfp #3 using a protoplast integration method. This work suggests that protoplast integration with dsRNA possibly generate significantly reduced gene expression in B. bassiana and the reduction is quite stable over generations which provide easy of functional study for fungal mode of action.

Two infamous ant species, red imported fire ant (RIFA, Solenopsis invicta) and tropical fire ant (TFA, S. geminata), originated from South America are invading and harming many countries around the world. Among them, RIFA, known as one of the 100 world’s worst invasive alien species, has been discovered in South Korea; in Busan (Gamman CT) in 2017, and subsequently in Busan (Hutchison CT), Pyongtaek, Incheon and Daegu in 2018. With these relentless introduction of RIFA, South Korea is not safe no more from these ants and faces a risk of settlement. Our research goal is tracing the country origin of invasive populations of RIFA and TFA based on molecular epidemiological approach. We are conducting both microsatellite and genome-wide SNP analyses in order to derive a complementary interpretation. These two methodologies are expected to result in improved resolution in population genetics analysis. Nevertheless, this work needs collaboration of many countries belonging to the range of origin and invasion of RIFA and TFA. We will briefly introduce our research project and progress of collaboration research.

The purpose of this paper was to investigate the comparison of balance and muscle strength between dominant and non-dominant legs in adults. Thirty adults in their 20s participated in this study. The dominant and non-dominant legs were selected based on the dominant hands of the target. The subject's muscle strength of legs was measured with Nicholas MMT, and the balance was measured with BIO-Rescue. We compared the dominant and non-dominant legs based on the results. The result, indicated no statistical difference on balance and muscle strength between dominant and non-dominant legs(p>.05). The results of this study will be helpful in setting the effective treatment direction and treatment level, and in controlling posture, balance and motor function.

To date, there are no protocols optimized to the effective separation of spermatogonial stem cells (SSCs) from testicular cells derived from mouse testes, thus hindering studies based on mouse SSCs. In this study, we aimed to determine the most efficient purification method for the isolation of SSCs from mouse testes among previously described techniques. Isolation of SSCs from testicular cells derived from mouse testes was conducted using four different techniques: differential plating (DP), magnetic-activated cell sorting (MACS) post-DP, MACS, and positive and negative selection double MACS. DP was performed for 1, 2, 4, 8, or 16 h, and MACS was performed using EpCAM (MACSEpCAM), Thy1 (MACSThy1), or GFR α1 (MACSGFRα1) antibodies. The purification efficiency of each method was analyzed by measuring the percentage of cells that stained positively for alkaline phosphatase. DP for 8 h, MACSThy1 post-DP for 8 h, MACSGFRα1, positive selection double MACSGFRα1/EpCAM, and negative selection double MACSGFRα1/α-SMA were identified as the optimal protocols for isolation of SSCs from mouse testicular cells. Comparison of the purification efficiencies of the optimized isolation protocols showed that, numerically, the highest purification efficiency was obtained using MACSGFRα1. Overall, our results indicate that MACSGFRα1 is an appropriate purification technique for the isolation of SSCs from mouse testicular cells.

Microenvironments surrounded with various extracellular matrix (ECM) components can decide specifically the fate of spermatogonial stem cells (SSCs) and integrin heterodimers recognizing directly ECM proteins play an important role in transporting ECM-derived signals into cytoplasm, resulting in inducing a variety of biological functions such as cell attachment, self-renewal and differentiation. However, to date, studies on type of integrin heterodimers expressed functionally on the undifferentiated SSCs derived from mouse with hybrid strain remain unclear. Therefore, we tried to investigate systematically what kind of integrin heterodimers are expressed transcriptionally, translationally and functionally in the SSCs derived from testis of hybrid (B6CBAF1) mouse. For these, magnetic activated cell sorting (MACS) using Thy1 antibody was used for isolating SSCs from testis, and real-time PCR or fluorescence immunoassay was conducted for measuring transcriptional or translational level of integrin α and β subunits in the isolated SSCs. Subsequently, antibody inhibition assay was conducted for confirming functionality of presumed integrin heterodimers. As the results, transcriptional levels of genes encoding total 25 integrin subunits were quantified, 7 integrin α (α4, α6, α7, α9, αV, αL and αE) and 2 integrin β (β1 and β5) subunit genes showed significantly increased transcriptional up-regulation, compared to the other integrin subunit genes. In contrast, integrin α3, α5, α10 and α11, and integrin β2, β3, β4 and β7 were weakly transcribed. When translational levels of the integrin α subunits showing high transcription level (α4, α6, α7, α9, αV, αL and αE) were measured, significantly strong translational up-regulation of integrin α6, α7, α9, αV and αL subunit genes were detected, whereas integrin α4 and αE subunit genes were weakly. In case of integrin β subunit, β1 evaluated more expression than β5. Based on these results, we speculated that the undifferentiated SSCs derived from B6CBAF1 mouse might express integrin α4β 1, α6β1, α7β1, α9β1, αVβ1 or αVβ5 on plasma membrane. Subsequently, the hybrid strain SSCs showed significantly increased adhesion to fibronectin, laminin, tenascine-C and vitronectin and functional blocking of integrin α4β1, α6β1, α9β1, and αVβ1 or αVβ5 in SSCs significantly inhibited attachment to fibronectin, laminin, tenascin-C and vitronectin, respectively. Accordingly, we could identify that the hybrid (B6CBAF1) mouse-derived SSCs had integrin α4β1, α6β1, α9β1, αVβ1 or αVβ5 on plasma membrane. Moreover, this information will greatly contribute to constructing non-cellular niche supporting self-renewal of SSCs in the future.

Insect-killing fungi have high potential in pest management. A deeper insight into the fungal genes at the whole genome level is necessary to understand the inter-species or intra-species genetic diversity of fungal genes, and to select excellent isolates. In this work, we conducted a whole genome sequencing of Beauveria bassiana (Bb) JEF-007 and characterized pathogenesis-related features and compared with other isolates including Bb ARSEF2860. A large number of Bb JEF-007 genes showed high identity with Bb ARSEF2860, but some genes showed moderate or low identity. The two Bb isolates showed a significant difference in vegetative growth, antibiotic-susceptibility, and virulence against Tenebrio molitor larvae. When highly identical genes between the two Bb isolates were subjected to real-time PCR, their transcription levels were different, particularly in heat shock protein 30 (hsp30) gene which is related to conidial thermotolerance. In several B. bassiana isolates, chitinases and trypsin-like protease genes involved in pathogenesis were highly conserved, but other genes showed noticeable sequence variation within the same species. Given the transcriptional and genetic diversity in B. bassiana, a selection of virulent isolates with industrial advantages is a pre-requisite, and this genetic approach could support the development of excellent biopesticides with intellectual property protection.

Korean horn beetles, Allomyrina dichotoma (Linnaeus, 1771) reared in many local farms are suffering from a fatal viral disease as reported in 2015, and recent sequencing analysis revealed that the virus is very closely related to Oryctes rhinoceros nudivirus. In a nationwide investigation, it was concluded that the virus can transmit vertically from an infected adult to the offspring by indirect manner, and 70 to 77% of young larvae dead in early stage were diagnosed as this virus positive. Here, we report for the establishment of on-site diagnosis method for the viral disease using loop-mediated isothermal amplification (LAMP) assay. A. dichotoma larvae were assayed for the OrNV infectivity with LAMP primers targeting OrNV_gp102 gene. To evaluate the LAMP specificity, two bacterial pathogens, Bacillus thuringiensis and Serratia marcescens, causing disease in A. dichotoma were tested along with OrNV. Only from the OrNV-infected larvae the reaction resulted positive. Also, to avoid DNA extraction process, the LAMP assay used diluted hemolymph directly and 50-fold dilution was set for diagnosis standardization and convenient on-site application for infected larvae from local farms. The LAMP diagnosis is fast and convenient for nontechnical user in the field and expected to stop this foreign virus spread all over the country.

Incursions of red imported fire ant (RIFA), Solenopsis invicta into Korea have been increasing. After a first interception of a colony of S. invicta on Gamman pier, Pusan port while intensive surveillance by Animal & Plant Quarantine Agency (APQA) in September 2017, three more RIFA colonies have been found in sea port piers of Pyeongtek, Incheon and Pusan cities. The social forms and mitochondrial DNA haplotypes of the intercepted RIFA colonies were analysed by allelic discrimination assay (peptide nucleic acid probe based RT-PCR) of Gp-9 gene and mt-DNA fragment of 768 bp, which is part of the Cytochrome oxidaseⅠ gene. The colony on Gamman pier, intercepted in Sep. 2017 was previously reported as a haplotype 5 (H5) of mitochondrial DNA and a social form of polygyne. The colony on Hutchison pier of Pusan port, intercepted in June 2018 were confirmed as a H22 haplotype and a monogyne. Those different social forms show different origins of each colonies. Those on piers of Pyeongtek and Incheon ports, also found in 2018 were confirmed samely as H22 and monogyne. However, it could be putatively assumed that those two colonies were differently introduced via different container cargoes, considering those colonies were found in container yards of distantly located different sea ports. More genetic variation analyses using diverse sets of molecular markers such as microsatellites, genome-wide single nucleotide polymorphisms, etc. in nuclear gens are being proceeded for more exact introduction routes (origins).

농업인구의 고령화 및 감소에 따른 노동력 부족으로 농업 생산 부분은 기계화 및 자동화를 비롯한 다양한 기술 도입을 요구하고 있다. 국내 과수 재배 작물 중 하나인 사과는 연간 10회 이상의 병해충 방제작업이 이루어지고 있어 병해충 방제를 위한 농약 살포 작업의 노력과 방제 비용, 작업자의 약제 노출 등을 줄이고 병해충 방제 효율을 높이는 방제 작업 시스템이 필요하다. 본 연구는 사과원 미세 분무 약제 살포 자동화 설비 구축을 위해 사과 과수원 내 미세 분무 살포 시스템을 설치하고 국·내외 산 노즐별 사과 잎의 농약 부착 정도를 비교하여 효과적인 노즐을 선발하기 위해 수행되었다. 국·내외 산 노즐별 사과 잎의 농약 부착 정도는 큰 차이를 보이지 않았으나 잎의 앞면 보다 뒷면의 농약 부착 정도가 낮게 나타나는 결과를 보였다.

벼메뚜기는 분류학적으로 메뚜기목(Orthoptera), 메뚜기과(Acrididae)에 속하는 곤충으로 작물에 피해를 주는 해충으로 인식하고 있지만 예로부터 단백질 보충을 위하여 채집하여 튀기거나 볶음요리 로 이용해 왔다. 벼메뚜기는 갈색거저리, 쌍별귀뚜라미 등과 함께 식품공전에 등록되어 있으나 1년 1세대로 가을철에 채집하여 이용하는 실정으로 공급확대에 한계가 있다. 최근 우리나라에서 벼메뚜기를 사육하고자 하는 농가와 다양한 용도로 가공을 통해 이용하려는 수요가 점차 증가하고 있으나 연중사육기술이 개발되지 않아 대량사육하는 농가가 거의 없는 실정이다. 이에 벼메뚜기의 연중대량사육기술 뿐만 아니라 인공사료 및 자동화 사육시설 개발에 따른 생산비 절감 기술 개발이 필요한 실정이다. 벼메뚜기의 먹이로는 여름작물로 옥수수, 수단그래스, 겨울작물로 밀, 보리 등의 볏과작물이 있으며 많은 양의 생엽과 발육에 적합한 작물의 선발 및 볏과작물의 통곡실 가루를 이용한 인공사료의 개발로 먹이공급 부족 시 15~20일 급여 가능하며, 벼메뚜기 사육시설로 단독형, 연결형, 절충형 등의 사육시설을 개발하여 연결형 사육시설의 수확량과 생존율이 가장 높았으며, 예취 급이 노동력이 단독형의 16% 소요되었다.

The continuous emergence of chemical-mediated residual toxicity and insect resistance have enforced the regulation of synthetic pesticides. Synthetic pesticides with novel mode of actions could be developed to overcome these issues, but as an alternative biopesticides with more efficacious control activity could be developed by the advanced technology. In pest management, entomopathogenic fungi have high potential in reducing pest population to an economic threshold, and some of isolates have been commercialized. However still a novel application strategy needs to be considered for successful industrialization. An insight on fungal genes in whole genome and transcriptome levels is necessary to understand the role of genes in pathogenesis and genetic diversity of fungal genes. Herein, we have identified the genetic differences of entomopathogenic fungi using whole genome sequencing of Beauveria bassiana (Bb) and tried to understand the interaction between fungus and insect using RNA-seq. We have obtained the whole genome sequence of Bb JEF-007 using PacBio sequencing technology and compared the transcriptomes of Bb JEF-007 and bean bug, Riptortus pedestris before and after the fungal infection using Hiseq 2000 system.

Entomopathogenic fungi have been known as promising candidates for biological control of insect pests. Recently, researchers consider the fungal thermotolerance in formulations and field applications. In this study, we investigated the production of thermotolerant Isaria javanica and I.fumosorosea conidia through grain-based solid cultures and exposure to light stress. As results, of the ten grain substrates, Italian millet, rice, perilla seed and sesame, rice, sorghum produced highly thermotolerant conidia in the strains. The two strains were exposed to a light stress and showed enhanced thermal stability compared to control, when exposed to 45°C for 2 hours. This work suggests that heatresistant entomopathogenic fungal conidia can be produced by grainbased solid cultures and exposure to light stress.

Japanese pine sawyer beetle, Monochamus alternatus Hope (Coleoptera: Cerambycidae) is considered as a serious pest in pine trees. To develop an eco-friendly strategy to manage this forest insect, we collected entomopathogenic fungi from Korean soil and assessed their virulence against the adults of the insect in laboratory conditions. As a result, two isolates with conidial suspension (1.0×107conidia/ml), showed 87% and 90% mortality 12 days after fungal treatment, respectively. We assessed the potential of the fungi-derived destruxin and protease as additives to the fungal isolates, and they showed insecticidal activity via feeding and spraying treatments. Finally, we produced fungal conidia in massive solid cultures and formulated wettable powders, and now studying optimal conditions of oil-based formulation with two isolates. Based on these results, we are evaluating the control efficacy of the fungal agents against M. alternatus in field conditions.

Pesticide application in agriculture provides significant benefits such as protection from disease, prevention of harmful insects, and increased crop yields. However, accurate toxicological tests and risk assessments are necessary because of many related adverse effects associated with pesticide use. In this review, we discuss and analyze residual pesticides contained in livestock feed in Korea. A pesticide residue tolerance standard for livestock feed has not been precisely established; so, risk assessments are required to ensure safety. Standards and approaches for animal criteria and appropriate methods for evaluating residual pesticides are discussed and analyzed based on technology related to animal product safety in Korea. The safety of livestock feed containing pesticides is assessed to establish maximum residue limits relative to pesticides. Analysis of residual pesticides in milk, muscle, brain, and fat was performed with a livestock residue test and safety evaluation of the detected pesticide was performed. Efficacy of organic solvent extraction and clean-up of feed was verified, and suitability of the instrument was examined to establish if they are effective, rapid, and safe. This review discussed extensively how pesticide residue tolerance in livestock feed and hazard evaluation may be applied in future studies.

본 연구는 기존에 재배된 바 없었던 암대극을 새로운 관상식물로 개발하고자, 절화 특성 평가를 실시하였다. 2017년 3월과 4월, 제주도에 자생하고 있는 암대극을 채취한 후, 수분흡수, 절화수명, 노화 과정 등 절화수명 특성을 평가하였다. 또한 sucrose 농도, 절화길이, pH 농도, 저장온도에 따른 절화 특성을 조사하였다. 본 실험은 국립수목원 유용식물증식센터의 Phyto-garden system(12h photoperiod, 25℃, 70% RH)과 실온(23℃, 44% RH)에서 수행하였고 2~3일 간격으로 수분흡수량, 생체중, 절화수명을 측정하였다. 실험 환경에 따른 연구에서 실온(14.4일) 보다 phyto-garden system(42.4일)의 절화 수명이 약 3배 더 길었고, 절화길이에 따른 연구에서는 절화수명이 20cm, 15.2일, 40cm, 17.4일로 나타났다. Sucrose 농도에 따른 연구에서는 처리에 따른 절화수명 증진 효과는 없었고, pH에 따른 연구의 절화수명은 pH 5, 15.6일, pH 6, 13.4일, pH 7, 13.1일, 증류수, 14.8일이었다. 저장온도에 따른 실험에서, 절화수명은 4℃, 83일, 10℃, 41.2일, 15℃, 35.5 일, 20℃, 17.4일, 실온, 14.4일이었는데, 온도가 감소할수록 절화수명이 길어짐을 알 수 있었다. 결론적으로, 암대극은 비교적 광도와 습도가 높은 환경과 절화길이가 길고 저장온도가 낮을수록 수명이 오래 유지되는 것을 알 수 있었다.

암대극(Euphorbia jolkinii Boiss.)은 우리나라 제주도 및 남부 해안가 암석지에서 자생하는 대극과(Euphorbiaceae)의 다년생 초본식물이다. 등잔모양꽃차례가 황록색으로 3월부터 5월까지 긴 기간 동안 개화하기 때문에 새로운 관상식물로 개발할 가치가 높다. 본 연구에서는 암대극을 재배화하고자 우선적으로 제주도의 자생지 환경과 개체군 분포 특성을 조사하였다. 제주도 전역의 해안을 따라서 암대극 개체의 전수조사를 실시한 결과 40여 지점에 걸쳐 약 14,817개체가 자생하는 것으로 확인되었다. 그 중 제주도 본섬의 동쪽에 위치한 우도에 1,075개체(전체의 7.3%), 북서쪽에 위치한 비양도에 952개체(6.4%), 남쪽에 위치한 가파도에 589개체(4.0%)가 조사되었다. 암대극은 주로 제주도 본섬 남동부 해안가 암석지대에 집중적으로 자생하는 것으로 확인되었고 가파도, 우도, 비양도의 경우에도 섬 동쪽지역에만 분포되어 있었다. 암대극의 생육은 자생지에 따라 차이가 있었다. 초장, 줄기 길이, 줄기 직경은 꽃 크기와 정의 상관관계를 보였다. 모든 지역에서 꽃이 피는 시기는 3월 말로 비슷하게 개화가 시작되었고, 6월 초에는 대부분의 꽃에서 종자가 성숙하여 탈리되었다. 암대극의 주변 식생은 갯개미자리, 돌가시나무, 순비기나무, 갯강활 등 30여 종이 분포되어 있었고, 초본이 80%를 차지하는 데에 반해, 목본은 20%로서 초본식물이 주를 이루었다. 토양의 pH는 8.3~9.4의 범위에 있었으며, 염분 농도는 0.003~0.017%로 나타났다. 유기물 함량은 중문에서 1.9%, 가파도에서 1.4%, 표선면에서 1.0%로 나타났고, CEC는 가파도에서 10.7cmol+/kg, 표선면에서 5.6cmol+/kg, 중문 지역에서 1.3cmol+/kg로 나타났다. 전기전도도는 중문, 가파도, 표선면에서 각각 0.47dS/m, 0.19dS/m, 0.16dS/m으로 나타났고, Cl-은 57.45mg/kg, 63.01mg/kg, 8.12mg/kg으로 나타났다. 중문에서 유효인산은 검출되지 않았다.

Development and fecundity of the foxglove aphid, Aulacorthum solani (Hemiptera: Aphididae) were investigated at ten constant temperatures (photoperiod: 15L:9D) from 2.5 to 30℃ on soybean (Glycine max) leaves. The nymphs couldn’t emerge to adults at 2.5 and 30℃. The lower development temperature threshold and thermal constant of development completion estimated with a linear development model in nymph were 5.02℃ and 131.2 degree-days. The lethal temperatures were estimated as 33.9 and 32.5℃ with Lactin 2 and Logan 6 non-linear models, respectively. Mean generation time (from 78.4 to 11.8 d) decreased with increasing temperatures (from 7.5 to 27.5℃). The highest net reproductive rate (77.4) was observed at 20℃. The highest intrinsic rates of population increase (0.282) and shortest population doubling times (2.07 d) were recorded at 25℃.