Japanese pine sawyer, Monochamus alternatus, is the main pest that mediates pine wilt nematode, Bursaphelenchus xylophilus, that causes serious damage to pine forests. In this study, we studied the strategy to control M. alternatus using entomopathogenic fungi. The fungi were collected from soil by an insect-baiting method and two fungal isolates (Metarhizium anisopliae JEF-197 and JEF-279) showed high virulence against M. alternatus. The Metarhizium isolates were evaluated for insecticidal activity against M. alternatus by spray treatment on live pine trees and wintering trees, and the M. anisopliae JEF-197 showed high insecticidal activity. In addition, the interaction of fungi and M. alternatus were analyzed by RNA-seq. This result can contribute to the development of insect control agents using entomopathogenic fungi.

The global biopesticide market was estimated to become about 4% of the total crop protection market in 2015, mainly due to variability of their efficacy, narrow spectrum or difficulties in long-term storage. Therefore, many people focus on overcoming these issues as a big trend. Suggested solutions include the investigation of synergy between microorganisms, the use of genetic engineering, improving the pesticide life shelf, etc. As a result, biopesticides market has grown by more than 17% over the last decade. In this context and aiming to develop new entomopathogenic fungi–based pest management tools, we constructed a fungal library by isolating insect pathogenic fungi from soil. A total of 581 isolates belonging to 35 species were isolated and characterized. Beauveria bassiana was the most abundant, representing 38.55% of the total strains, followed by Metharizium anisopliae (22.55%) and bubillosa (8.6). …% of the total isolates were highly virulent against Tenebrio molitor killing most of the treated insects in 2 to 3 days.

The longhorned tick, Haemaphysalis longicornis, is one of the vectors of severe fever with thrombocytopenia syndrome virus (SFTSV) in human. The use of pyrethroid insecticides induced pest resistance and environmental residual toxicity. Here in this work, our interest was given to the selection of highly virulent fungi against longhorned tick. A total of 101 fungal pathogens were assayed by dipping the nymph stage of ticks into a conidial suspension. Interestingly of the several species, one species showed high virulence and mycosis were observed in 7-15 days. Highly virulent strains were selected, and semi-field experiments were conducted. As a result, the control efficacy of the isolate was over 80% at 30 days of treatment. This work suggests that entomopathogenic fungi could be used to effectively control longhorend ticks.

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.

누에는 약제, 면직, 식용, 동물 사료 등에 활용도가 높아 동아시아에서 오랫동안 사육되어 왔다. 뽕잎을 먹고 자란 누에가 사료를 먹고 자란 누에보다 생장주기가 일정하고 더 건강하게 생장한다는 것은 잘 알려진 사실이다. 또한 최근의 연구에 따르면 동물 장내 미생물이 동물의 면역과 생장에 큰 역할을 한다고 알려졌다. 이러한 사실로부터 뽕잎에 존재하는 미지의 미생물이 누에 생장과 면역에 관여할 수 있다는 가설을 세우고 이를 확인하기 위한 연구를 수행하였다. 뽕잎 내생균을 분리하기 위해 뽕잎을 5mm 크기로 잘라 표면살균 후, 배지에 치상하여 자라나오는 세균을 분리하여 동정하였다. 흥미로운 점은 100개 이상의 뽕잎 절편으로부터 동일한 세균이 검출되었으며 계통학적 분석을 통해 동정한 결과, Pseudomonas syringae pv. syringae(Pss)로 확인되었다. 또한, 뽕잎을 섭식한 누에의 장 속과 분변에서 이 균의 존재를 확인하였다. 누에에 병을 일으키는 곤충병원성 곰팡이에 대한 Pss의 항진균 효과를 확인하기 위해 곤충병원성 곰팡이인 Beauveria bassiana, Cordyceps militaris와 식물병원성 곰팡이 Botrytis cinerea, Fusarium graminearum을 대상으로 대치배양을 통해 균사 생장 억제 정도를 확인하였다. 그 결과, 식물병원성 곰팡이의 생장 억제효과는 관찰되지 않았으나 곤충병원성 곰팡이의 생장이 50% 이상 감소되었다. 특히, 다량의 포자를 생성하여 곤충에 치명적인 병을 야기하는 B. bassiana의 균사 생장이 C. militaris에 비해 더 크게 감소하였을 뿐 아니라 Pss의 배양 추출물을 처리하였을 때 B. bassiana의 포자 발아가 지연되었다. 앞으로 누에 장 속에 안착한 Pss가 곤충병원성 곰팡이에 대한 면역력과 누에 생장에 미치는 영향을 확인해 볼 계획이며 이를 통해 건강하고 안정적인 누에 사육에 활용될 수 있는 미생물 제제를 만들고자 한다.

The development of entomopathogenic fungi has received increasing interest as part of integrated pest management strategies as biocontrol agents. It is reasonable to assume that entomopathogenic fungi might produce secondary metabolites modulating juvenile hormone for their survival against defense mechanisms of host insects. In this study, acetone extracts of 189 entomopathogenic fungi cultured on unpolished rice medium were screened for their juvenile hormone antagonist (JHAN) activities using the yeast-two hybrid system. Among 14 extracts showing JHAN activities, extract of the F-145 showed high level of insecticidal activities against both Plutella xylostella and Aedes albopictus. This isolate was identified as Lecanicillium attenuatum. These results suggested that the Lecanicillium attenuatum could be useful for development of eco-friendly insecticides.

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 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.

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.

The population of managed honeybees has been dramatically declining the recent past in worldwide. N. ceranae causessignificant detriment to honey production and results in economic losses critically. In our knowledge, Fumagillin is theonly antibiotic approved for control of nosemosis in honeybees. In this study, to select isolate with anti-Nosema activityagainst N. ceranae. Entomopathogenic fungi cultural filtrates were screened using in vitro polar tube germination assay.These fungi cultural filtrates were used to evaluate the safety of honeybees and their inhibition of nosemosis. As a result,P. marquandii 364 and Pochonia sp. 60 showed inhibitory activity on the growth of N. ceranae in honeybees and didnot significantly affect the survival rate of honeybees. These may be employed as antibiotic agents and a good featureto be used in the development of new biocontrol agents of nosemosis.

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.

Entomopathogenic fungi have been widely studied for their potential as the effective biological control agents. Theyproduce variety of secondary metabolites with insecticidal activities, and it is reasonable to assume that entomopathogenicfungi might produce secondary metabolites modulating juvenile hormone for their survival against defense mechanismsof host insect. In this study, Acetone extracts of 189 entomopathogenic fungi cultured on unpolished rice medium werescreened for their juvenile hormone antagonist (JHAN) activities using the yeast-two hybrid system. Among them, 14extracts showed high level of JHAN activities and their insecticidal activities against Aedes albopictus were investigated.

The diamondback moth, Plutella xylostella is one of the world’s major pests. Economic cost to control this pest wasestimated between US$1.3 billion and US$2.3 billion based on management costs. Conservative estimate included yieldloss caused by 5% diamondback moth was estimated US$4 billion-US$5 billion. P. xylostella was managed by chemicalinsecticide such as organophosphates, carbamates and pyrethroids. But insecticide resistance which is caused by repeatedapplication makes it difficult to control this pest. For environmental friendly control of diamondback moth, entomopathogenicfungi could be used as alternative. We conducted bioassay to select high virulent isolate to larva of diamondback mothwith forty six entomopathogenic fungi which were isolated from soil samples by insect-bait method. As a result of bioassaytwelve isolates was selected as candidate. We investigated control efficacy of these twelve isolates with potted Chinesecabbage at laboratory and greenhouse.

Entomopathogenic fungi have been used to control pest as alternative to chemical pesticide. To kill the pest entomopathogenicfungi penetrate cuticle of pest, reach the hemocoel and utilize nutrient of host pest. Finally fungi kill the host by consumingthe host nutrient and physically damaging the tissues. But these process of fungi to control pest is needed so much timeand this point is a disadvantage for fungi. Therefore we studied other application method of fungi to control pest. Weconducted behavior test of beet armyworm to Isaria fumosoroseus which is high virulent against beet armyworm. Adultof the beet armyworm avoided oviposition at Chinese cabbage treated with I. fumosoroseus compare to control and otherhigh pathogenic isolate, Metarhizium anisopliae and this repellency of I. fumosoroseus lasted for 5days in greenhouse.Behavior of larvae to I. fumosoroseus also investigated with choice and non-choice test. Third to fifth instar larvae detectedand avoided fungi. Repellent behavior of larvae to fungi was more noticeable in younger larvae. This result may be usedto prevent the infestation of moth in crop production.

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.

The green peach aphid, Myzus persicae, is one of the most important pests of vegetable crops worldwide. This species cause direct damage by feeding on plant nutrients and indirect damage as transmits many virus vectors. The control of aphid has relied on the use of chemical insecticides and their intensive use over many years has led to the development of resistance. To surmount or avoid these problems, we screened the entomopathogenic fungi against green peach aphid and evaluated their virulences. Several entomopathogenic fungal isolates were selected with high virulence to green peach aphid. Additionally, the antimicrobial activity of the selected fungal isolates was tested and analyzed to phytopathogens. Consequently, these entomopathogenic fungi would be used effectively for dual control agents for the green peach aphid and phytopathogen.

Spodoptera exigua (Hübner) is an important pest with wide host range including cabbage, beans, potato etc. We evaluate the virulence of Beauveria bassiana ARP14, B. bassiana GHA, and Isaria japonica with concentration of 1×108 conidia/mL against larval stage of S. exigua using glass scintillation vial. Lethal median time (LT50) of these three fungi was not found significantly different in both fourth and second instar. Infection rate was higher in fourth instar than second instar. Infection rate was highest by B. bassiana ARP14 (84.0%) followed by B. bassiana GHA (63.6%) and I. japonica (45.8%) in fourth instars of S. exigua. Our results suggest that B. bassiana ARP14 can be a new component of integrated pest management of S. exigua.

Entomopathogenic fungi, widely distributed in nature, have been developed as biopesticides of agricultural pests. However, little is known about isolation method for obtaining a variety of highly virulent strains from the soil. In this work, the previously established method using mealworm (larvae of Tenebrio molitor) as a baiting system was used to efficiently isolate fungal strains from soil, and the isolated fungal library in Korea was established. To isolate entomopathogenic fungi, soil samples from 10 regions of Korea such as Iksan, Imsil, Gunsan, Yeosu, Suncheon, Gwangyang, Jinju, Hamyang, Boseong and Jangseong were collected. Of the collected soils, 21 and 34 isolates of entomopathogenic fungi were isolated from Iksan and Gunsan, respectively. The isolates were classified as genera of Metarhizium, Beauveria and Isaria. This work can suggest an effective insect pathogenic fungal isolation method

Entomopathogenic fungi have been known as promising candidates in biological control. However, a large number of conidia with higher stability should be produced for industrialization. In this work investigated the possibility of several grains for the production of thermotolerant Isaria javanica and I. fumosoroseus conidia. The fungal conidia were produced on ten grains for 14 days in Petri dish conditions. Of the ten granular substrates, millet, perilla seed and barley showed the high conidial production, 15.7 ± 8.4, 17.3 ± 5.2 and 13.5 ± 1.9 × 109 conida/g in I. javanica and 15.9 ± 0.6, 15.0 ± 0.5 and 15.0 ± 0.7 ×109 conida/g in I. fumosoroseus, respectively. However, the two strains showed relatively low thermal stability regardless of substrates, when exposed to 45℃ for 1, 2 and 3 hours. This work suggests a possible substrate for conidial production of entomopathogenic fungi with low cost.