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.

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.

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.

Due to their environmental and economical consequences, invasive species have become a major concern worldwide. Among them, the yellow-legged hornet, Vespa velutina, is a keen hunter of domestic honeybees. They use olfactory cues from the prey to assess food information. In this study, we investigated the attraction behavior of V. velutina to honeybee pheromone under outdoor conditions. Nine honeybee pheromones were tested and compared in order to find the best attractant. When testing specific compounds, the honeybee queen pheromone, homovanillyl alcohol, proved highly attractive.

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.

Entomopathogenic fungi are expected to play a role as great biological control agents in the global bio-pesticide market in the future. The first step is the collection of fungal isolates and it should be a platform for the development of highly effective biopesticides. In this work, we constructed a fungal library using a mealworm pathogenecity-based fungal collection method and further characterized some isolates with high virulence. A phylogenetic tree was generated, and of the isolates 17 isolates’ biological features were characterized, such as morphology, virulence against several insect, stability of conidia for heat, production of biologically active materials, such as enzymes. This work reports an attractive entomopathogenic fungal library including the information of effective isolates in pest management.

Many turfgrass pests, Ectinohoplia rufipes, Exomala orientalis and Popillia quadriguttata, cause serious damage to grass fields in Korea. In this study, control of turfgrass pests using entomopathogenic fungi were investigated using Protaetia brevitarsis seulensis as an alternative insect. In a preliminary experiment, some Metathizium sp. showed ca. 60% virulence after 7 days exposure to the entomopathogenic fungi. Compared to the contact-exposure, overall a spray treatment showed lower control efficacy against P. brevitarsis seulensis. Furthermore, mortality of various developmental stages of P. brevitarsis seulensis was investigated under laboratory conditions. This work suggests that when treated with fungi mortality of P. brevitarsis seulensis was closely related with developmental stage. These results provide a basic information for the control of P. brevitarsis seulensis using entomopathogenic fungi

Composite resins are developed as restorative materials to improve esthetics and mechanical properties. To improve the physical properties of resin material, resin filler have to be added. However, no imaging method is adopted for resin filler distribution. Optical coherence tomography (OCT) is a optical imaging technique to delineate microscopic structures within biological tissue. The OCT application to dental composites resin and its filler is not described yet. So, this new and advanced optical method is needed for clinical application for evaluation of dental composite resin. To analyze the spatial distribution of dental composite resin and to evaluate the resin restoration in cavity, frequency domain optical coherence tomography (FD-OCT) was used for their analysis. Resin restored tooth was prepared. For morphological observation, serially sectioned teeth, conventional X-ray taking and micro computed-tomography (CT) images were compared with OCT images. The experiment has done to evaluate the success of the resin restoration using 3 dimensional structure OCT image. In this research, OCT is evaluated as a new technique to image resin restoration. The evaluation of resin restored tooth was performed by OCT. Inappropriate restoration such as marginal adaptation, large porosities, internal integrity and poor contour could be detected. Resin filler also could be checked by OCT. The distribution, number, regularity and size of resin filler can be differentiated from several commercial products. Considering the characteristics of the OCT, it can be used to evaluate the defects of resin restoration, resin filler distribution, and internal integrity between resin material and tooth structure. The OCT can be considered to be a new and advanced method for the evaluation of resin restorations.

In organic agriculture, various cover-crops have been used to control weeds. In this study, we have investigated the effect of Vicia tetrasperma (L.) Schred. (Eolchigiwandu) which is native to on major insect pests of pepper. Redpepper seedlings at 8 leaves stage were transplanted in 20th May 2009 into experimental field located in the farm of the department of agricultural biology, Suwon, Korea. A cover-crop cultivation plot was compared with a control plot mulched by black plastic-film. Density of aphids and damaged fruits by oriental tobacco budworm, Helicoverpa assulta, were surveyed ten times from 21 days after transplanting (DAT) to 82 DAT. Fifty and forty pepper plants were sampled to count the density of aphids and damaged fruits by oriental tobacco budworm per a plant, respectively. In current study, three aphid species namely, cotton aphid, Aphis gossypii, green peach aphid, Myzus persicae, potato aphid, Macrosiphum euphorbiae were collected. Overally, the density of aphids on pepper in the control plot was higher than the cover-crop plot. Especially, aphid density increased up to 67.44±26.8 in early stage of control plot, whereas aphid was not found in cover-crop plot. The rate of fruit damage by oriental tobacco budworm was significantly higher in the cover-crop plot than the control plot in early stage of pepper, however damaged fruit rate of pepper in the control plot was significantly higher since the middle of July.

Background : Methicillin-Resistant Staphylococcus aureus (MRSA) is a multidrug-resistant (MDR) strain. Especially, MRSA is developing resistance to available antibacterial agents and causing complications in the treatment of infections related to skin, soft tissue, respiratory, bone, joint, and endovascular disorders. Therefore, antibacterial agent combination therapy appears to be a useful option, particularly in developing countries where antibiotic availability is limited. (+)-Usnic acid (UA) is uniquely found in lichens, and is especially abundant in genera such as Usnea and Cladonia. UA has antimicrobial activity against human and plant pathogens. Therefore, UA may be a good antibacterial drug candidate for clinical development. Methods and Results : In search of a natural products capable of inhibiting this multidrug-resistant bacteria, we have investigated the antimicrobial activity of UA against MRSA. In this study, the effects of a combination of UA and permeable agents against MRSA were investigated. For the measurement of cell wall permeability, UA with concentration of Ethylenediaminetetraacetic acid (EDTA) was used. In the other hand, Sodium azide (NaN3) was used as inhibitors of ATPase. These results suggest that the antibacterial effect of UA was potentiated by membrane-binding agents and ABC transporter-inhibiting agents, implying that antibacterial activity is associated with damage of the cell wall and inhibition of ATPase function by UA. Conclusion : UA and in combination with EDTA and NaN3 could lead to the development of new combination antibiotics against MRSA infection. The results of this study appear to be promising, and they are expected to enhance the use of natural products as drugs.

Background : Ginseng, one of most famous traditional oriental medicines, has been known for a number of pharmacological properties including anti-tumor, anti-diabetic, anti-fatigue, anti-stress, anti-oxidative, and anti-aging effects. 20(S)-Protopanaxadiol (PPD), a intestinal metabolite of ginsenosides, is one of the active ingredients in ginseng. In this study, we investigated the synergistic anticancer effect of 20(S)-PPD and temozolomide (TMZ) and the mechanism of 20(S)-PPD on glioblastoma cells. Methods and Results : We examined cell viability and the morphological changes of C6 cells after treatment of 20(S)-PPD and TMZ. 20(S)-PPD showed a potent antiproliferative activity against C6 cells by triggering apoptosis. 20(S)-PPD-induced apoptosis was characterized by a dose-dependent mitochondrial damage. 20(S)-PPD and TMZ had a synergistic effect in increasing mitochondrial damage via caspase 3 activation. Conclusion : These results revealed an unexpected mechanism of 20(S)-PPD and TMZ, triggering a mitochondrial-mediated apoptosis in C6 cells. Our findings encourage further studies of 20(S)-PPD as a promising chemopreventive agent against glioblastoma.