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 serious emergence of chemical-mediated residual toxicity and insect resistance have been enforced the regulation of synthetic pesticides. Future decisions to select more realistic control options probably depend on the speed of technological development in chemical and biological pesticides. Now, a strategic collaboration between synthetic pesticides and biopesticides has been progressed, such as distribution and R&D in collaboration and M&A for obtaining microbial resources. Recently registered microbial pesticides are entomopathogenic fungi in pest management. A concept of e-biopesticide could be properly combined with digital agriculture and accelerate the use of biological control agents in the future farming.

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.

봄철 천식환자 가정집 9곳에서 실내 공기를 포집하여 부유진균의 농도 및 다양성을 조사하였다. 조사된 천식환자 가정집의 평균 부유진균 농도는 거실 143 CFU/m³, 방 142 CFU/m³ 이었으며 외기보다 낮았다. 이는 다중이용시설 실내공기질 기준법의 권고기준 기준치 500 CFU/m³를 초과하지 않았음을 보여준다. 총 59균주를 분리하였으며 5속 33종으로 동정하였다. 이 중 병원균으로서 인체에 위해성을 가지고 있는 진균은 4속 12종이었다. 동정된 종 중 Cladosporium crousii, Penicillium maximae, Aspergillus protuberus는 국내에 보고되지 않은 미기록 진균 종으로 확인하였다. 이 중 A. protuberus는 국외에서 인체에 질병을 유발시키는 균으로 알려진 진균이다. 이상의 결과는 천식환자 가정에서 진균의 관리는 정량적 수준에서뿐만 아니라 유해종을 파악하여 관리하는 정성적인 수준에서도 이루어져야 함을 제시한다. 본 연구는 봄철 천식환자 가정집에 존재하는 실내 공기 중 부유진균의 농도 및 다양성에 대한 최초의 연구이다.

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 aim of this study was to select various fungal strains indigenous to Korea that have the potential to produce cellulases, including filter paper activity (FPase), endo-β-1,4-glucanase (EG), and β-glucosidase (BGL). Among the 25 species of Ascomycetes and the 32 species of Basidiomycetes tested in this study, the Bjerkandera adusta KUC10565, Heterobasidion orientale KUC10556, Hyphoderma praetermissum KUC10609, and Trichoderma harzianum KUC1716 all exhibited remarkably high FPase activity. In addition, the T. harzianum KUC1716 showed high levels of EG and BGL activity. This strain has been selected for further study because of their enzymatic potential.

To evaluate effects of ligninolytic enzyme type on the mycelial response and ligninolytic enzyme production during interspecific interactions among wood-rotting fungi, 4 fungal strains, Trichophyton rubrum LKY-7, Trichophyton rubrum LSK-27, Pycnoporus cinnabarinus, and Trichoderma viride, were selected. Regarding ligninolytic enzyme production, LKY-7 secreted laccase and manganese peroxidase (MnP), P. cinnabarinus secreted only laccase, and LSK-27 secreted only MnP in glucosepeptone medium, while T. viride did not produce any ligninolytic enzymes. In the co-culture of LKY-7 with P. cinnabarinus, the formation of aerial mycelium was observed and the enhancement of laccase activity owing to interspecific interaction appeared to be very low. In the co-culture of LKY-7 and P. cinnabarinus with LSK-27, a hypha-free clear zone was observed, which resulted in deadlock, and increased laccase or MnP activity was detected at the interaction zone. The interaction responses of LKY-7, P. cinnabarinus, and LSK-27 with T. viride were characterized by the formation of mycelial barrages along the interface. As mycelial barrages were observed at the T. viride territory and no brownish pigment was observed in the mycelial barrages, it is suggested that laccase and MnP are released as part of an offensive response, not as a defensive response. The co-culture of P. cinnabarinus with T. viride lead to the highest enhancement in laccase activity, yielding more than 14-fold increase in laccase activity with respect to the mono-culture of P. cinnabarinus. MnP activities secreted by LKY-7 or LSK-27 was generally low in interspecific interactions.

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.

The two-spotted spider mite, Tetranychus urticae, and gray mold disease caused by Botrytis cinerea, are importantpest and plant disease. To control these, many people have been relied on chemical methods for a long time. However,their continuous use has resulted in resistance of pest and disease. To surmount or avoid these problems, we evaluatedantifungal activity of the selected fungi with high virulence to mite to explore the potential for the dual control of notonly T. urticae but also B. cinerea. To maximize the use of spores and cultural filtrates, the virulence to mite was evaluatedusing culture products, and effective culture condition was investigated against blastospore production and antifungal activity.Consequently, the 2 fungal isolate selected in this study were confirmed to have diverse potential and would be usedeffectively for dual control agents against the two-spotted spider mite and plant diseases

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.

Lignosus rhinocerotis, commonly known as Tiger Milk Mushroom, has been long extolled for its medicinal properties and used for treatment of asthma, cough, fever, cancer, liver-related illnesses, joint pains and as tonic. The history of usage for Tiger Milk Mushroom dates back to almost 400 years, ago but there were no records of scientific studies done due to inavailability of sufficient samples. Even when there were samples collected from the wild, the supply and quality was inconsistent. With the advent of cultivation success of one of the most utilized species of Tiger milk mushroom (L. rhinocerotis) in 2009 (known as TM02), scientific investigation was done to validate its traditional use and to investigate its safety for consumption, biochemical and biopharmacological properties. Preclinical toxicology evaluations showed that TM02 did not induce adverse effects on fertility. It also poses neither teratogenic effects nor genotoxicity. The no-observed-adverse-effect level (NOAEL) dose of the TM02 in 180-day chronic toxicity study is 1000 mg/kg which is equivalent to 162 mg/kg human dose, anticipated dose for a 60 kg adult is ~10g. The effective dosage of TM02 is as low as 0.5g per day for an adult. There is therefore, a huge margin for the safe consumption of TM02. TM02 is proven to be effective in supporting the lung and respiratory health, immune health, maintenance of joint health (caused by inflammation), improving vitality and stamina and improving the quality of life for cancer patients. Recent scientific findings have shown that TM02 contains various active components such as the polysaccharides-protein complexes, β-glucans, proteins, immuno-modulatory proteins all of which could play possible roles in rendering significant therapeutic properties such as anti-inflammatory, immuno-modulatory, anti-proliferative, anti-oxidative, neurite outgrowth stimulation etc. The revelation of the genome, transcriptome, proteome of L. rhinocerotis by MMRG has provided valuable insights into the biomolecule discovery and provided the foundation for future research and exploitation of L. rhinocerotis in pharmacological and functional food applications. These data forms a valuable foundation for future research in the exploitation of the L. rhinocerotis in pharmacological and industrial applications.