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.

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.

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.

Beauveria bassiana is widely studied for its potential as biopesticide. However, little is known about the factors to influence genetic diversity among isolates. In this work, we aimed to study the gene diversity of 42 isolates to figure out the impact of sequences variability on various biological features. The accumulated data showed no correlation between sequence variability and the fungal geographic localization. Meanwhile, weak to strong correlations have been obtained between the sequence diversity of various genes and the studied biological features. Interestingly, Biotrophy-associated gene 2 gene variability was correlated with all the studied biological features, being an important marker to determine isolates of interest for biological control. Therefore, the obtained data would serve as a database to focus on interesting genes related to improving or screening highly virulent isolates as biological control agents.

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.

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 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.

Silverleaf whitefly, Bemisia tabaci is a worldwide agricultural pest that cause serious damage to crops. However, since this insect developed resistance to variety of chemicals, alternative control strategy needs to be studied. In this study, we aimed to evaluate the potential of entomopathogenic fungi as biological control tools against this pest. We developed a dipping method in laboratory conditions to screen insecticidal activity of different fungal species and isolates against the nymphal stage of whitefly. The highly virulent isolates were characterized and 2 isolates were further selected based on their high conidial productivity and thermotolerance. Finally, several grain substrates were used to confirm the conidial productivity and thermotolerance of the selected isolates, revealing optimum conidial production when the fungi were cultured on millet grains. Both isolates could be used in further millet grain-based formulations to control the whitefly in fields.

Red mite, Dermanyssus gallinae is one of the serious pests, damaging to the egg production in chickens. Moreover, the overuse of chemical insecticides caused pest resistance and environmental residual toxicity. Therefore, this work provides a screening method to select entomopathogenic fungi as a candidate of environmentally safe control agent, having high miticidal activity against D. Gallinae. The virulence test was conducted using a spraying method. Of the several species, Beauveria species showed high virulence and mycosis were observed. Some isolates were produced on eight cereal grains in Petri dish conditions. Millet, perilla seed and barley showed the high conidia production. Conidia of the isolates produced on millet and rice showed high thermal stability, when exposed to 45℃. Based on these results, this work suggests that entomopathogenic fungi could be used to control the D. Gallinae.

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.

갈색날개매미충은 기주 범위가 넓고, 효과적 방제가 어려운 해충으로 알려져 있으며, 향후 더 큰 피해를 유발할 수 있는 해충으로 지목되고 있다. 경북지역의 안동과 의성에서 3월부터 9월까지 4개 지점을 선정하여 갈색날개매미충의 월동란이 산란된 월동기주를 중심으로 월동란 부화 시점 및 부화 후 약충에서 성충까지 그리고 다시 월동란 산란까지의 패턴을 모니터링하였다. 각 지점의 환경조건 및 수목류 군락의 조성은 조금씩 차이가 있으나 충분한 산란흔이 있었다. 1령 약충은 5월 중하순부터 하순까지 나타났고 산수유나무, 매실나무에서 가장 먼저 확인되었으며, 이후 수목류에서의 밀도증가 양상은 매실나무에서 더 빠르게 진행되었다. 그리고 2령 약충 까지는 기주식물에 인접한 수목류로의 이동한 모습도 일부 관찰되었다. 그러나 6월 중순경에는 3령 약충의 기주식물 주변 초본류로의 이동이 관찰되었으며, 월동란에서의 부화 후 1회차 이동분산으로써, 주로 상부에서 하부로의 점프를 통한 이동으로 판단된다. 이는 새순 부위가 경화된 월동 기주식물에서 경화되지 않은 초본류의 새순으로의 이동으로써, 특정한 초본류에 집중되지 않고 넓은 범위에 산재하여 분포하였다. 4령 약충 시기에는 주변 특정 초본류와 목본류 새순에 집중적으로 분포하는 패턴을 보이기도 하였다. 7월경에는 성충이 관찰되었으며, 이는 2회차 분산이 진행되는 시점으로 판단된다. 이후 8월 중순까지 지속해서 밀도 감소가 이뤄지던 월동 기주에서의 밀도 감소 상태가 증가 추세로 전환되었다. 그리고 9월 상순경에는 2018년 월동기주에서의 성충밀도 증가와 교미 행동 및 1년생 가지에서의 산란흔이 관찰되었다. 산란흔의 집중도는 산수유나무와 매실나무가 다른 수목류보다 높았으며, 이는 모니터링이 진행된 모든 지점에서 공통된 패턴임을 확인할 수 있었다.

Gonggeom-ji is designated wetland protected area by the Ministry of Environment of Korea in 2011 because of it’s high biodiversity and historic value. It contains a reservoir, paddy and forest site which has diverse niches for insects. Three transect lines of 50m were designated. Data were collected in May, October, November in 2017 represented as seasons for spring, summer and autumn respectively. Quantitative methods were conducted along each transect line by sweeping and pitfall trap. Based on this study total of 1079 individuals of insect fauna were collected, representing 170 species in 60 families and 8 orders. The highest diversity, richness and evenness index were observed in the forest site in May(4.77, 8.6 and 0.91 respectively) and the highest dominance index was observed in the forest site in November(0.64). The highest similarity index was observed in the reservoir site in May and August(0.519).

Longhorned tick, Haemaphysalis longicornis (Ixodida: Ixodidae) is one of the vectors of severe fever with thrombocytopenia syndrome virus (SFTSV) in human. The tick occurs in mostly grass fields, and the use of pyrethroid insecticides induced pest resistance and environmental residual toxicity. Particularly the use of chemicals near residential areas where persons live become a big issue, so environmentally safe control agents need to be considered. Here in this work, our interest was given to the selection of highly virulent fungi against H. longicornis. A total of 101 fungal pathogens collected from mountainous areas were assayed by a dipping the nymph stage of ticks into a conidial suspension (1×107conidia/ml). Interestingly among several species, one species showed high virulence and mycosis were observed in 7-15 days. Most of the selected isolates produced a large amount of conidia in Italian millet, rice and millet with thermotolerance at 40-45°C. Based on these results, we selected a couple of isolates with high virulence against H. longicornis and they could be used for the control of the ticks after the set-up of a practical application strategy in fields by optimizing fungal colonization in soil and phyllosphere.

Thrips is one of the serious insect pests in many economic crops, in which general managements rely mainly on chemical insecticides. However, the overuse of chemicals led to the development of insect resistance and environmental residual issues. Therefore, thrips management requires the consideration of alternative strategies which are less harmful to the environment and working on different target points. In this work, we isolated entomopathogenic fungi from soil, and identified them with morphological and molecular biology methods, followed by a preliminary virulence assay against Tenebrio molitor larvae. Selected fungal isolates were adjusted to 1×107 conidia/ml for an indoor virulence assay against thrips adults, and highly virulent isolates were added to a thrip-pathogenic fungal library. Biological characteristics of efficacious isolates were investigated comparing to a previously commercialized fungal isolate. This entomopathogenic fungal library could be used as a valuable resource for developing effective strains to control thrips in agricultural fields.

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.

Whitefly has a broad host range of more than 600 species, and 40 biotypes have been reported worldwide. Among the various ecological types, B and Q are the most problematic. Especially, Biotype Q, which causes serious damage in the green house, mediates more than 40 kinds of viruses, including Tomato yellow leaf cull virus (TYLCV). The development of resistance to whitefly, adverse effects on non-target organisms, and strengthened legal regulations are making chemical control difficult in application. For these reasons, the use of alternative strategies is needed to suppress the Biotype Q whitefly population. Entomopathogenic fungi were investigated and developed to control hard-control insect pest, such as whitefly. These fungi are natural pathogen of their invertebrate host and contribute to the regulation of their host population in the environment. In addition, these agents are typically harmless to humans and non-targeted organisms and do not adversely impact the environment. Therefore, in this study, we screened pathogenic fungi against whitefly using our entomopathogenic fungal library, and virulence tests were conducted with conoidal suspension (1×107conidia/ml) in laboratory conditions. As a result, 10 isolates showed 80-100% mortality at 5 days after treatment. We will analyze the characteristics of these fungal isolates and evaluate their potential as insecticides against whitefly. The selected isolates against the high mortality of whiteflies can be effectively used in integrated pest management programs for whiteflies.

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.