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℃.

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.

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 a heat stress. And they showed enhanced thermal stability compared to control, when exposed to 45°C for 2 hours. This work suggests that heat-resistant entomopathogenic fungal conidia can be produced by grain-based solid cultures and exposure to light stress.

Melon thrips, Thrips palmi Karny (Thysanoptera: Thripidae) is a serious insect pest of various crops in the world. The management of this pest mainly have relied on chemical agents. However, the overuse of chemical agents for long times causes insect resistance, and negative effects on environment. Therefore, alternative control methods, such as biological control, that are less harmful to the environment and have different mode of action are needed to control this pest. In this study, we isolated entomopathogenic fungi from Korean soil, and characterized them via morphological and molecular identifications and pathogenicity check against Tenebrio molitor larvae. The isolated fungi were subjected to virulence assay against T. Palmi with a conidial suspension of 1×107 conidia/ml in laboratory conditions. This entomopathogenic fungi library would be a good resource to select effective strains for the control of T. palmi in fields.

Entomopathogenic fungi play an important natural role in regulating their insect host populations, and their ecology was also associated with plant and soil. These microorganisms have been living by reacting to insect, plant and environmental factor. The advanced bioinformatics technology such as next-generation sequencing and RNA sequencing has revolutionized in understanding of entomopathogenic fungi. Recently studies provided a lot of information on evolutionary relationships and virulence-related characteristics. We are starting to know where these microorganisms from is, and how they live in nature. The bioinformatics technology will give us further our understanding of the natural roles of these fungi in nature.

Little consideration has been given to the insect epizootiology in pest management. High production cost of fungalinsecticides might be a technical barrier. Application of the fungal insecticides to unfavorable conditions might result inlower performance in pest management. Herein this work, we suggest ecological biocontrol considering long-term colonizationin nature rather than quick pest control. Fungal biopesticides should be frequently combined with chemical pesticides infields, particualrly spatial combination strategy rather than temporal combination. Additionally other important technicalaspects need to be strongly considered, such as economic downstream process, effective control and environmentally safe,so finally proposing e-biopesticide. Lastly R&D system needs to be clearly and efficiently organized for high performance.Week-based summary and reporting system, weekly report and project file are main system in our laboratory which hasour lab work more effectively and efficiently, consequently resulting in tech-transfer to an industry and re-investment.

The entomopathogneic fungal colonization has become an interesting issue in ecological pest management. Plant-orientedfungi induce plant growth promotion and act as natural control agents against insect pest. Herein this work, we evaluatedthe ability of fungal colonization on bark, branches and leaves of pine tree (Pinus densiflora) to select optimal candidatesfor controlling Japanese pine sawyer (Monochamus alternatus), which is the pine wilt disease vector in Korea. As a result,some Beauveria and Metarhizium isolates could colonize on the substrates of P. densiflora and showed pathogenicity againstM. alternatus. This result suggests that the selected entomopathogenic fungi are possibly candidates for long-term managementof M. alternatus in pine tree forest.

Japanese pine sawyer, Monochamus alternatus HOPE (Coleoptera: Cerambycidae) are considered as important pine forestinsect pests in several countries which transmits the pine wood nematode, Bursaphelenchus xylophylus. In this study, wetried to control Japanese pine sawyer using entomopathogenic fungi to prevent the spread of pine wilt disease in Korea.Several entomopathogenic fungi were exposed to the adults of Japanese pine sawyer and mycosis was observed in themembranous cuticles after 6 days. In particular, two isolates showed high virulence against the adults. The selected 2isolates were sprayed to the adults at 1×105, 1×106 and 1×107 conidia/ml. In the concentration of 1×107 conidia/ml, thesurvival rates of insects were 40% and 47%, respectively. Based on these results, the possibility of controlling the Japanesepine sawyer was confirmed.

Chemical pesticides have been used for persimmon pest control, however the overuse caused insect resistance andenvironmental toxicity. Herein this work, we investigated the biological potential of eco-friendly organic pesticides (biologicals)and entomopathogenic fungi in controlling persimmon pests, Stathmopoda masinisaa and Riptortus pedetris. A botanicalmixture of Chinese scholar tree, goosefoot and subtripinnata extract, Bacillus thuringesis NT0423 and one isolate of Beauveriabassiana showed high insecticidal activity against the pests in laboratory conditions. The selected botanical extract andmicroorganism were combined with a chemical pesticide (buprofezin+dinotefuran WP) and were assessed their controlefficacy in persimmon fields. Compared to the three times of chemical spray, the combination of more than one timeof chemical spray and the selected biologicals controlled the persimmon pests with an acceptable levels. Consequentlyone time of biological spray can be integrated to the spray program where only chemicals are strongly considered.

The hard tick, Haemaphysalis longicornis (Ixodida: Ixodidae) is one of the vectors of severe fever with thrombocytopeniasyndrome virus (SFTSV) causing in humans, however little consideration was given to the biological control of the pest.We evaluated three bioassay methods, such as spray, dipping and dropping. As a result, the dipping method was themost efficient way to measure the fungal virulence. Finally, we constructed a Haemaphysalis longicornis pathogenic fungallibrary to further facilitate the resources to be used as potential biological control agents. A conidial suspension of 1×107conidia/ml was exposed to the ticks by dipping. We confirmed that some isolates were highly virulent to the ticks. Thisresults indicated that some fungi could be used to effectively control the Haemaphysalis longicornis.

Many entomopathogenic fungal isolates received interests for controlling insect pests. But very few numbers of isolatesreached agricultural fields as commercial products. In this study, we investigated the feasibility of grain substrates forconidial production, focusing on conidial productivity and thermotolerance. Next to efficiently use the produced conidiafor spray application, conidia need to be separated from the grain substrates. Isaria javanica and I. fumosorosea conidiawere solid-cultured on ten kinds of cereal grains. Solid-cultured grains were packed in a screen bag and immersed inwater with a variety of surfactants at 100 ppm, such as Silwet, CO-2.5, CO-12, LE-7, PE-61, TDE-3, Tween 20 or Tween80. High levels of conidial release were observed on LE-7, TDE-3 and Tween80 treatments. This work suggests thatthe screen bag can be used to separated conidia from the cultured grains with the help of surfactants.

A genus of entomopathogenic Beaveria bassiana has been widely used in pest management, however little studies havebeen given to its virulence-related genes. To identify the roles of virulence genes, AtMT-induced random mutants weregenerated and followed by localization study with TAIL-PCR. Two genes, Complex I intermediated-associated protein30 (CIP30) and Autophagy protein 22 (Atg22), were predicted as virulence-related genes in B.bassiana JEF-007. To validatethe a possible relationship between two genes and fungal virulence, hpRNAi was performed. A hpRNAi plasmid wasconstructed as a model system to knock down of egfp gene in egfp-expressing B. bassiana transformant. A Real-TimePCR showed the knock down of egfp gene expression via hpRNAi. The CIP30 and Atg22 genes in B. bassiana willbe validated by the established RNAi technique. This work can be a strong platform for the functional genetics in B.bassiana.

풀무치 (Locusta migratoria) (Orthoptrea: Acridiade)는 전 세계적으로 작물 생산에 심각한 문제를 야기하는 돌발 해충이다. 그러나 우리나라 의 경우 풀무치를 방제하기 위한 방제제 및 적용에 대한 연구는 미흡한 실정이다. 본 연구에서는 풀무치에 병원성을 갖는 풀무치병원성 진균 라이브러 리를 구축하였으며, 풀무치 방제에 이용 가능한 생물학적 방제제로서의 가능성을 평가 하였다. 먼저 갈색거저리 유충-baiting 시스템을 이용하여 다양 한 지역에서 채집된 토양에서 곤충병원성 진균을 발굴 하였다. 풀무치 병원성 검정을 진행하기 위하여 국립 농업 과학원에서 풀무치를 분양 받았으며, 고체 배양된 곤충병원성 진균을 곤충 사육 상자에 처리하여 (2 g/ box), 풀무치 약충 (3-4령충)에 대한 곤충병원성 진균의 병원성을 평가 하였다. 그 결과 곤충병원성 진균 처리 3-7일차에 풀무치의 머리, 복부, 다리 표면에서 진균이 증식하는 mycosis를 확인 할 수 있었다. 특히, Metarhizium anisopliae, M. lepidiotae, Clonostachys rogersoniana에서 높은 병원성이 나타나는 것이 확인 되었다. 확보된 34개의 풀무치병원성균주의 특성을 파악 하기 위하여 열안정성 및 포자생산성을 확인 한 결과, Paecilomyces, Purpureocillium 균주가 다른 균주에 비해 열에 대한 높은 안정성안 나타나는 것을 확인 하였으며, 대부분의 균주에서 1 × 108 conidia/gram 이상의 포자수를 생산 하는 것을 확인 하였다. 또한 온실 조건에서 비교적 병원성이 높았던 M. anisopliae 고체 배양된 균주를 토양에 처리하여 병원성을 확인한 결과, 85.7%의 높은 방제효과를 확인 할 수 있었다. 본 실험을 통하여 풀무치가 이동하면서 토양에 정착된 곤충병원성 진균에 접촉되어 치사 될 수 있을 것으로 판단되며, 효과적인 풀무치 방제가 가능 할 것이라고 판단된다.