Insect killing fungus Beauveria bassiana has been widely studied as a biological control agent. However, many studies have been focused on lab or field-based management. Herein this work, comparison of three B. bassiana strains was investigated under a molecular level. The whole genome sequences of ERL836, JEF-007 were analyzed by PacBio (35.5 Mb of ERL836 and 36.5 Mb of JEF-007) and ARSEF2860 referenced from GenBank (33.7 Mb). To compare the three strains, virulence, thermotolerance and chemical resistance were assayed. The transcriptomes of non-infecting B. bassiana and infecting B. bassiana against western flower thrips were analyzed using RNA-seq. This work can provide that genome features, functions, morphology and gene expression could be different under the molecular level, even if in the same species.

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.

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.

In order to accumulate the ecological information of Drosophila suzukii for export negotiations of domestic strawberry, we weekly monitored occurrence of D. suzukii at three strawberry fields located in Gok-seong, Dam-yang and Hwa-sun using the best combination of trap and attractant from November 2017 to May 2018, the period of strawberry production. Monitoring was carried out at three sites (a hill located 150 meter distant to greenhouse, outside greenhouse and inside greenhouse) in Gok-seong. Drosophil suzukii was trapped at three short periods from November to the beginning of December, from the end of December to the beginning of January, and from April in a hill. At outside greenhouse, D. suzukii was trapped at two short periods from November to the early December and the beginning of May. In inside greenhouse, it occurred once after May 24th. Gok-seong data can be summarized as more occurrence for longer period as the distance to greenhouse increases. Dam-yang occurred once in a short period inside greenhouse and hill, respectively on May. In case of Hwa-sun, D. suzukii did not trapped in whole trapping sites during trapping period, except for the surrounding grape and peach cultivation areas at the end of November. Summarized, no occurrence periods are from November 20th, 2017 to May 14th, 2018 inside greenhouse; from December 11th, 2017 to May 3rd, 2018 outside greenhouse; and from January 8th, 2018 to April 12th, 2018 on a hill. In connection with strawberry culture, the D. suzukii tends to occur at the beginning and the end of the strawberry production period. We expect this result can serve as a helpful data for export negotiation for quarantine.

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.