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.

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.

Inflammation is an important protective response mechanism that occurs against microbial invasion or injury. However, excessive inflammation may lead to cause of morbidity and mortality in diseases. The activated macrophages plays a vital role in inflammatory response by stimulation of lipopolysaccharide (LPS) and tumor necrosis factor-α (TNF-α). This activation further damages the host by inducing certain pro-inflammatory mediators such as nitric oxide (NO), interleukin-1β (IL-1β), interleukin- 6 (IL-6), TNF-α, inducible nitrous oxide (iNOS) and cyclooxygenase (COX-2). Flavonoids are bioactive compounds with potential effects as anti-cancer, anti-inflammation, anti-viral and anti-bacterial activities. Polymethoxyflavones (PMFs) are unique to citrus plants which are of specific interest owing to their biological effects that includes lipoprotein metabolism and anti-inflammatory activity. Sinensetin is one of the PMFs having five methoxy groups on the basic benzo-γ-pyrone skeleton with a carbonyl group at the C4 position. Sinensetin have been known for exerting various pharmacological activities including anti-angiogenesis, anti-diabetic and anti-inflammatory activities. However, there are no studies focused on the anti-inflammatory effects of sinensetin on skeletal muscle cells. In the present study, we investigated the antiinflammatory effect of flavonoids isolated from Sinensetin on the production of pro-inflammatory mediators mediated by nuclear factor-kappa B (NF-κB) by inhibition of signal transduction in LPS - induced L6 skeletal muscle cells.

Cryopreservation of bovine embryos is used to efficiently implant surrogate mothers. It has been widely accepted that high lipid content in the oocyte interrupts its survival during freeze-thaw cycles. Serum component in the culture medium is thought to increase the embryo`s lipid contents. Conversely, L-carnitine stimulates lipid metabolism by transporting long chain fatty acids into the mitochondria. Objective of this study was to analyze the effect of L-carnitine supplementation in IVM medium and defined IVC medium on the development, lipid contents and the cryosurvival of bovine IVF embryos. 0.0, 1.5, 3.0 and 6.0 mM L-carnitine was supplemented in IVM medium, respectively (IVM-LC 0.0, LC 1.5, LC 3.0 and LC 6.0). Development rate from the 2cell to the morula stages was higher in IVM-LC 3.0 groups than those of IVM-LC 6.0 (p<0.05). But there were no significant differences among the other groups in the blastocyst rates and lipid content results. When 0.0, 1.5, 3.0 and 6.0 mM L-carnitine were supplemented in IVC medium (IVC-LC 0.0, LC 1.5, LC 3.0 and LC 6.0), development competence was not significantly different between those embryos. Lipid contents of embryos treated L-carnitine (IVC-LC 1.5, 3.0 and 6.0) were significantly lower than embryos of non-treated group. L-carnitine was supplemented 0.0, 1.5, 3.0, 6.0 mM during IVM and 3.0 mM during IVC (LC 0.0 - 3.0, LC 1.5 – 3.0, LC 3.0 – 3.0, LC 6.0 – 3.0) and cryosurvival of blastocysts confirmed after freezing-thawing. There were no significant differences on development, but LC 3.0 – 3.0 was significantly lower lipid contents than other groups. And LC 3.0 – 3.0 had better survival rates and hatched rates of blastocysts than LC 0.0 – 0.0. In conclusion, supplementation of L-carnitine in defined IVC medium decreases lipid contents. And L-carnitine supplementation improves cryosurvival and developmental ability of bovine IVF embryos.

The early-onset familial Alzheimer's disease (EOFAD/ FAD), the less common type of Alzheimer's disease (AD) currently affects a vast number of individuals worldwide. This type is being inherited as an autosomal dominant fashion. Missense mutations on Amyloid precursor protein (APP) and Presenilins 1 and 2 (PSEN1 & PSEN2) are known as major genetic factors in FAD. Conversely, missense mutations on microtubule-associated protein tau (MAPT) are also thought to involve. Up to date, several triple-transgenic animal models with muted forms of the human APP, PSENs and MAPT have been reported. Compared to other animals, canines are more emotional and their disease signs can be easily diagnosed. This attempt was to develop a triple transgenic canine model for the AD. We have obtained the coding sequences of APP, PSEN1 and MAPT from Dana-Farber/Harvard Cancer Center DNA resource core at HMS and incorporated several common AD mutations. The transgenic construct is composed of hNSE (ENO2) promoter-driven three AD genes fused together with modified 2A sequences. It was transfected into the canine fetal fibroblasts which were then used to perform somatic cell nuclear transfer (SCNT). The viable transgenic embryos were obtained after in vitro culture and the GFP was detected. In this study, we have successfully produced viable triple transgenic canine cloned embryos using SCNT technique. These transgenic canine embryos will be further developed into canines with FAD. The transgenic canines will be a good candidate in the AD research field.

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.

2018년 현재까지 농림축산검역본부 호남지역본부로 수입된 식물은 220종 17,030건으로 우드펠렛(18%), 주정박(8%), 페스큐 건초(8%), 알파파건초(8%), 귀리건초(6%), 혼합건초(5%), 티모시건초(5%), 라이그래스건초(5%), 목화씨(사료용)(4%), 소나무원목(3%) 등의 순이다. 이와 같이 호남지역은 비재식 사료류(건초 등) 수입비중이 높으며 이는 전국 수입 건수의 약 67%에 달한다. 검역해충은 795건 검출되었으며 이 중 관리해충 294건, 잠정규제해충 146건, 비검역해충 307건 등이다. 관리해충 중 Carpophilus obsoletus가 210건(72%), 잠정규제해충 중 Cryptoletes ferrugineus가 56건(39%)으로 높은 검출율을 보였다. 또한 호남지역으로 수입 된 재식용 구근류는 생강 및 생강구근, 마늘구근이며 전국 수입 건수의 약 42%를 차지한다. 수입 된 생강 및 생강구근 91건 중 37건에서 해충(선충)을 발견하였고 Meloidogyne sp.(잠정규제해충) 검출로 9건이 폐기·반송되었다. 수입된 마늘구근 34건 중 29건에서 해충을 발견하였고 Carpophilus obsoletus(관리해충) 18건, Cryptoletes ferrugineus(잠정규제해충) 11건이 검출되어 소독하였다.

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.

본 연구에서는 국내 유통 중인 농산물 9품목(n = 578)에 대한 납과 카드뮴 함량을 조사하고 이들의 섭취로 인한 위해성을 평가하고자 하였다. 납과 카드뮴의 함량은 마이크로웨이브 분해 후 ICP-MS로 분석하였다. 조사대상 농산물의 납 평균 함량은 각각 보리 0.014 mg/kg, 완두콩 0.010 mg/kg, 강낭콩 0.008 mg/kg, 녹두 0.006 mg/kg, 파인애플 0.008 mg/kg, 살구 0.016 mg/kg, 매실 0.015 mg/kg, 자두 0.021 mg/kg, 대추 0.019 mg/kg이었고, 카드뮴 평균함량은 보리 0.017 mg/kg, 완두콩 0.004 mg/kg, 강낭콩 0.007 mg/kg, 녹두 0.005 mg/kg, 파인애플 0.001 mg/kg, 살구 0.002 mg/kg, 매실 0.002 mg/kg, 자두 0.002 mg/kg, 대추 0.003 mg/kg이었다. 모든 시료의 납, 카드뮴 함량은 EU, CODEX 및 국내 기준보다 낮은 수준이었다. 조사 대상 농산물에 대한 납, 카드뮴의 인체노출량을 산출한 결과, 납은 잠정주간섭취허용량(PTWI, 25 μg/kg b.w./week)의 0.067%, 카드뮴은 월간잠정섭취허용량(PTMI, 25 μg/kg b.w./month)의 0.28%이었다. 이상의 결과는 조사 대상 농산물의 납, 카드뮴 오염도와 이들의 섭취에 의한 위해성이 모두 낮은 수준이라는 것을 보여준다.

벼메뚜기의 인공사료 개발을 위하여 일본의 Konno식 먹이를 대조구로 하고, 28개의 배합비율별 인공사료를 제작하여 급이시험을 하였다. 시험결과, Konno식 보다 6주 후 1마리당 체중은 22개 처리에서 높았고 우화성공율은 23개 처리에서 높았다. 폐사율은 부화입식 후 45일까지 Konno식 대비 24개 처리에서 높았다. 1마리당 체중, 우화성공율, 폐사율을 종합해 볼 때 탄수화물원으로는 옥수수 건조잎 분말 또는 통밀가루와 단백질원으로는 2~5%의 탈지대두분 또는 어분의 배합비율이 벼메뚜기의 인공사료로서 가능성이 높았다.

Beauveria bassiana (Bb) is a potential biopesticide against western flower thrips. To understand molecular characteristics of two Bb isolates (ERL836 and JEF-007), whole genome sequencing and RNA-seq were performed. The whole genome of Bb ERL836 was sequenced (3,934,801,443 bp) using Pacbio sequencing and 15 contigs were analyzed. To understand the molecular mechanisms of pathogenicity in two Bb genome (before infection and after infection), next generation sequencing (NGS) was analyzed to compare transcriptomes. A result of differentially expressed gene (DEG) showed that 2,689 contigs were up-regulated and 2,619 contigs were down-regulated. Randomly selected five genes were validated to be highly up-regulated. This work can provide an understanding of the interaction between Bb and westetn flower thrips.

Japanese pine sawyer, Monochamus alternatus Hope (Coleoptera: Cerambycidae) in forest is considered as a serious forest pest in pine trees. In this study, we collected entomopathogenic fungi from soil and assessed their virulence against the adults of the inset using a spray method in laboratory condition. Two entomopathogeinc fungi isolates showed high virulence against the adults. In the concentration of 1.0 × 107conidia/ml, the mortality rate of the adults were 53% and 60%. In semi-field condition, one isolate showed a virulence of 60% against the adult. Consequently, we confirmed the possibility of the fungal isolates in controlling the beetles. In near the future, we will investigate several factors which is possibly related to the control of the forest insect pest using entomopathogenic fungi in field conditions, given the importance of fungal formulation and practical application methods.

The hard tick, Haemaphysalis longicornis (Ixodida: Ixodidae) is one of the vectors of severe fever with thrombocytopenia syndrome virus (SFTSV) causing in humans, however little consideration was given to the biological control of the pest. Therefore, this study provides a screening method to select entomopathogenic fungi, having high virulence against H. longicornis. The virulence test was conducted by dipping method with a conidial suspension (1×107 conidia/ml). As a result, cadavers of H. longicornis infected with entomopathogenic fungi were obtained by this method. Based on this results, we selected several isolates having high virulence to H. longicornis. We suggest that, selected isolates in this study can be used for the control of H. longicornis.