최근 다양한 유기재배 작물의 뿌리를 가해하는 굼벵이류의 피해가 증가하고 있으나, 굼벵이류는 토양 내 발생하는 특징으로 인해 발생시기 및 그 종류에 대한 확인이 어려운 해충이다. 피해를 끼치는 굼벵이의 발생을 파악하기 위해 고구마 유기재배지에 페로몬 트랩을 이용하여 굼벵이의 성충의 종류 및 발생 동향을 조사하였다. 조사지는 무안 유기재배농가와 국립농업과학원 완주군 포장에서 이루어졌다. 3종의 풍뎅이 페로몬 루어를 유인제로 사용하였으 며 6월 초부터 8월 말까지 조사지에 트랩을 설치하여 포획된 풍뎅이를 수집하여 동정을 하였다. 유기재배포장에서 포획된 종은 큰검정풍뎅이, 콩풍뎅이, 청동풍뎅이, 녹색콩풍뎅이, 별줄풍뎅이 등의 풍뎅이와 흰점박이꽃무지 등이 주로 채집되었다. 유기재배 고구마포장에서 풍뎅이 발생소장을 조사한 결과 최대로 발생한 시기는 7월초였다.

Fumigation activity of Frankliniella occidentalis, western flower thrips, and phytotoxicity were evaluated with phosphine(PH3) and ethyl formate (EF) in the asparagus. Fumigation activity of PH3 and EF showed in the order of egg (64.7 mg·h/Land 132.9 mg·h/L), nymph (5.5 mg·h/L and 14.8 mg·h/L), and adult (3.8 mg·h/L and 14.7 mg·h/L) respectively at 5˚C in12 L desiccator. The adsorption of PH3 was 24% at 5˚C. However, the adsorption of EF increased sharply to 98% at 5˚C.PH3 showed 100% control for all development stage of F. occidentalis with 4g/m3 in 0.65m3 at 2˚C for 48h exposure. Thephytotoxicity was not found. The results revealed that the EF is highly absorbable in asparagus, so it is not suitable as afumigant, but PH3 is considered to be suitable as an alternative MB fumigant for control of the F. occidentalis in asparagus.

Ethyl formate (EF) and phosphine (PH3) is an alternative fumigant to methyl bromide (MB). For applying import sweet pumpkins in Korea, the efficacy of EF and PH3 was evaluated on Tetranychus urticae adult and egg stage which is pest in sweet pumpkins. The eggs of T. urticae were more tolerance than adults in both EF and PH3. When T. urticae eggs were treated with EF for 4 hrs, LCT99=107.63 mg/L and LCT99=45.37 mg/L at 5°C and 20°C, respectively. PH3 treatment at 5°C and 20°C for 24 hrs, eggs were controlled LCT99=49.44 mg/L and LCT99=17.23 mg/L, respectively. The mixed treatment of EF and PH3 showed no significant synergistic effect on T. urticae. However, EF (80 mg/L) and PH3 (4 mg/L) treatment did not cause any external phytotoxicity damages in sweet pumpkin even when treated with the maximum amount of fumigant at 5℃ and 20℃ for 24hrs.

Zika virus, mosquito-borne disease, caused by mosquitoes has been increased the importance. From March to September, twice a month from 7 different points (3 residential areas, 3 migratory bird sanctuary and 1 cattle shed) were collected using BG trap and BL trap. After identifying the mosquitoes collected, we confirmed the virus infection. Total 26,531 mosquitoes (6 genus 9 species) were collected, virus has been detected from the 3 species (Aedes vexans, Cuilex tritaeniorhynchus, and Amigeres subalbatus) of mosquitoes of them. It showed the highest peak in August, and then gradually decreased. The most common mosquito species was collected Aedes vexan (16,637) in the cattle shed.

Monochamus saltuarius is one of the insect vector of pine wilt disease with Monochamus alternatus. The present study aimed to investigate the dose to induce sterile and the effect of sterility according to the gender of M. saltuarius. As the electron beam irradiation dose increased, the hatchability, adult eclosion rate and longevity of the former mid and late maturation feeding stage of adults was slightly reduced. However, there was no difference between these two adults. In addition, female adults were more sensitive to the electron beam than the male adults. Hatchability of the F1 generation of ♂T × ♀N o r♀T × ♂N (T: 200 Gy electron beam irradiation, N: 0 Gy) was completely suppressed. As electron beam irradiation dose increased, the fecundity of female adults were slightly decreased. Results by comet assay showed that electron beam irradiation induced as dose increased DNA damage in M. saltuarius adults. These results suggest that SIT using electron beam may be useful for control of M. saltuarius.

Mealybugs (Hemiptera: Pseudococcidae) are major pests of a wide range of crops and ornamental plants worldwide, and represented by 56 species in Korea. Occurrence of, or damage by, the longtailed mealybug, Pseudococcus longispinus Targioni-Tozzetti has not been reported since 2002. However, for the first time in many years, P. longispinus was reported at Phalaenopsis orchids by Kim et al in 2015. Therefore, the present study aimed to investigate the distribution of P. longispinus which is usually found inside a greenhouse or a building where the host plants are. We have collected 129 mealybug samples at 254 sites from Korea. Among these, 27 of P. longispinus and 62 of an unidentified Pseudococcus species were found mainly in ornamental plants such as Heteropanax fragrans, commonly known as happy trees in Korea. Morphological characteristics of the unidentified mealybug were very similar to those of P. longispinus. However, the genetic variability in COI sequence comparison was significantly different between the two species. Further studies are needed to clarify the valid name for this unidentified species, and we are currently working on it.

Hypocreales entomopathogenic Beauveria bassiana and Metarhizium anisopliae are well-known biological control agents worldwide and have high potential in industrialization. However their thermo-susceptibility limits long-term storage under high temperature conditions and high insecticidal activity after application to target pests. Herein we isolated highly virulent isolates, B. bassiana JEF006 and JEF007 and M. anisopliae JEF003 and 004, and produced in three grains, as substrates for solid cultures, followed by thermotolerance assays to compare the potential of the three substrates. The JEF isolates were exposed to dry and wet heat at 50°C and overall conidia were more stable under dry heat condition rather than wet heat. Of the three grains, Italian millet was superior to the other grains in the production of thermotolerant conidia. Additionally Italian millet did not severely aggregated, which enabled air to penetrate into the substrate well compared to the sorghum and millet. JEF isolates were more thermotolerant when they were kept in oil conditions. The conidia of JEF 007 and 003 wild type and AtMT-based generating random mutants were subjected to SDS-PAGE. A significant relationship between conidial thermotoelrance and detected proteins was observed. This work suggests that Italian millet can be used as an effective substrate to produce more thermotolerant conidia, thus maintaining viability for long times under unfavorable environment and biological activity against target pests.

Bean bug, Riptortus pedestris is an agriculturally serious pest in north eastern Asian countries, damaging to several legumes and fruit trees. Chemical pesticides have been largely used to control the pest but it encounters insecticide-resistance and environmental toxicity issues. Alternatively different mode of action and environmentally sound pest management system can be found in entomopathogenic fungal insecticides. Herein we developed a platform to optimize the fungal production to express their maximum virulence against bean bug, by focusing on solid culture system for thermotolerance, formulation to select effective surfactants to carry the fungal conidia on the cuticles, and relationship between environmental abiotic factors and fungal mortality. First to produce highly thermotolerance fungal conidia, Beauveria bassiana and Metarhizium anisopliae isolates were cultured on several granular cereal substrates, which could be subjected to formulation process. Among the tested media, four media (millet, non-glutinous italian foxtail millet, glutinous italian foxtail millet, brown rice) were superior to the other grains in the spore production and thermotolerance. Next to efficiently deliver the fungal conidia on the cuticles of bean bug, total of six surfactant (CO-2.5, CO-12, LE-7, PE-61, TED-3 and Siloxane) was used to experiment. CO-12 was superior to the other surfactant in mortality of 100 ppm consistence. This work suggests that solid culture system and formulation and application should be seriously considered to reach an optimal level of mortality by inducing their maximum virulence.

Entomopathogenic fungi are facultative microorganisms, dwelling in soil or infecting host insects, and some of the genera have been used as biological control agents worldwide. Collection of fungal isolates should be a platform for the development of highly effective resources, thus 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 three was generated, and of the isolates 17 isolates’ biological features were characterized, such as morphology, spectrum of virulence, cultural characteristics, thermo-stability of fungi, 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.

ocust, Locusta migratoria (Orthoptera: Acrididae) is one of the outbreaking pests worldwide and such big occurrence was recorded in 2014, Korea, however little consideration was given to the management strategy of the pest. Herein we established a indoor locust-rearing system and constructed a locust-pathogenic fungal library to further facilitate the resources to be used as possible biological control agents. A locust colony was provided from the National Institute of Agricultural Science and Technology and reared in corn or barley plants at artificially manipulated rooms. The critical developmental stages, such as oviposition, hatching and mating were successfully proceeded. Entomopathogenic fungal granules were treated to the locust (2 g/rearing box), and in 5~7 days mycosis was observed in the membranous cuticles of head, abdomen and legs. In particular JEF-003 (Metarhizium anisopliae), JEF-186 (M. lepidiotae) and JEF-187 (Clonostachys rogersoniana) showed high virulence against the locust. A population of locust was exposed to the entomopathogenic fungal conidia-incorporated soil to investigate the possibility of the fungal isolation from natural soil, which resulted in the pathogenesis in 7~10 days in laboratory conditions. More than 80% of control efficacy was observed in the greenhouse trial of fungal granular application. This work suggests that locust rearing system was successfully established and entomopathogenic fungi can be used to control the migratory locust.

Entomopathogenic Beauveria bassiana and Metarhizium anisopliae are well-known biological control agents worldwide and have high potential in industrialization. However their thermo-susceptibility limits long-term storage under high temperature conditions and high insecticidal activity after application to target pests. Herein we isolated highly virulent isolates, B. bassiana JEF006 and JEF007 and M. anisopliae JEF003 and JEF004, and produced in three grains, such as sorghum, millet and Italian millet as substrates for solid cultures, followed by thermotolerance assays to compare the potential of the three substrates for thermotolerance. The JEF isolates were exposed to dry and wet heat at 50°C and overall conidia were more stable under dry heat condition rather than wet heat. Of the three grains, Italian millet was superior to the other grains in the production of thermotolerant conidia. Additionally Italian millet did not severely aggregated, which enabled air to penetrate into the substrate well compared to the sorghum and millet. JEF isolates were more thermotolerant when they were kept in oil conditions as carriers of an oil-based formulation. This work suggests that Italian millet can be used as an effective substrate to produce more thermotolerant conidia, thus maintaining viability for long times under unfavorable environment and biological activity against target pests.

Entomopathogenic fungi infect some arthropod pest and have been used for biological control. Some entomopathogenic fungi have high potential in insect pest management worldwide, and most of researches were given to Hypocreales order of Ascomycota, but little in Zygomycota, such as Entomophthorales in our country. We have identified some species belonging to Entomophthorales and investigated cultural features and ecology of Entomophaga aulicae. E. aulicae in sweet potato fields, where Aedia leucomelas was a dominant pest, had caused epizootics from 2002–2005. E. aulicae-infected A. leucomelas larva were mostly found from August to October and its occurrence was significantly related to the precipitation. E. aulicae were mainly observed in dead A. leucomelas larval populations (infection rate = 41.3% in 2002), rather than other lepidopteran pests. E. aulicae was morphologically and genetically identified and its virulence was characterized in laboratory conditions. Additionally, pest infection by Zoophthora radicans, Neozygites floridana and some Entomophthorales have been identified. On the basis with this information, we need research to predict the prevalence and to develop biological control agent using Entomophthorales including E. aulicae that contribute to regulation of host populations.

Antimicrobial peptides (AMPs) can be produced in mealworms. In this work, we integrated Bombyx mori (Bm) AMP, cecropin A to Beauveria bassiana ERL1170 by restriction enzyme-mediated integration method, which was confirmed by RT-PCR and an antibacterial activity assay. For the extracellular secretion of Bm cecropin A protein, the active domain of the cecropin A gene was tailed to the signal sequence of B. bassiana chitinase (Bbs). To exchange Bbs-cecropin A gene with egfp gene in pBARKS1-egfp, Bbs-cecropin A fragment was cut from pGEMBbs-cecropin A using XbaI/blunted and BamHI and ligated with cut pBARKS1-egfp using NcoI/blunted and BamHI, designated as pBARKS1-Bbs-cecropin A. After the transformation, transformants were grown on Czapek’s solution agar containing 600 μg ml-1PPT. Expression of Bm cecropin A was confirmed by RT-PCR. Strong clear zone was observed in the co-culture of the transformant D-6 and Bacillus subtilis on fourth strength Sabouraud dextrose agar 1 day after the culture at 25°C, whereas the wild type had no clear zone. This work suggests that Bm cecropin A can be efficiently produced in this mealworm-based fungal expression platform, thereby increasing the value of mealworms in the animal feed additive industry.

Bean bug, Riptortus clavatus Thunberg (Heteroptera: Alydidae) causes serious damage to Leguminosae. Herein an entomopathogenic fungal virulence assay system against bean bugs was established to construct a fungal database which can be used in integrated pest management (IPM). First to obtain as many bean bugs as possible at the same stage, host plant-preference and developmental synchronization of bean bugs were investigated. In the preference assay, five pairs of adults were infested in a plastic cage, where a pot of green bean, pea or cowpea was previously placed. The highest fecundity and the fastest development of bean bug was observed in the green bean cage. Secondly, in the synchronization experiment, eggs were collected from the cage of adults in 1, 3, 5 and 7 days after oviposition and transferred to a fresh cage with green beans. From the every 4 days of survey, similar stages of bean bugs were found in the cages with the oviposition for 1 and 3 days, rather than the longer times of oviposition. A fungal bioassay against bean bugs was conducted using the bean bugs from the above insect rearing system. Ten Beauveria bassiana isolates were cultured on quarter-strength Sabouraud dextrose agar (¼SDA) for 7 days at 25°C. Ten 4th instar of nymphs were placed on a cultured plate for 1 hour and tranferred to a fresh moisturized plate with grains of green bean. ERL836 isolate treatment showed the highest virulence and fungal mycosis was observed on the bean bugs. In conclusion, these results can be useful to establish an entomopathogenic fungal database for IPM.

Antimicrobial peptides (AMPs) can be produced in mealworms, currently being used as animal feeds, by the infection of genetically engineered-entomopathogenic fungi. In this work, we integrated Bombyx mori (Bm) AMP, cecropin A to Beauveria bassiana ERL1170 by restriction enzyme-mediated integration method, which was confirmed by RT-PCR and an antibacterial activity assay. For the extracellular secretion of Bm cecropin A protein, the active domain of the cecropin A gene was tailed to the signal sequence of B. bassiana chitinase (Bbs). To exchange Bbs-cecropin A gene with egfp gene in pBARKS1-egfp, Bbs-cecropin A fragment was cut from pGEM-Bbs-cecropin A using XbaI/blunted and BamHI and ligated with cut pBARKS1-egfp using NcoI/blunted and BamHI, designated as pBARKS1-Bbscecropin A. After the transformation, transformants were grown on Czapek’s solution agar containing 600 μg ml-1PPT. Expression of Bm cecropin A was confirmed by RT-PCR. Strong clear zone was observed in the co-culture of the transformant D-6 and Bacillus subtilis on fourth strength Sabouraud dextrose agar 1 day after the culture at 25°C, whereas the wild type had no clear zone. This work suggests that Bm cecropin A can be efficiently produced in this mealworm-based fungal expression platform, thereby increasing the value of mealworms in the animal feed additive industry.