Calliphora is a genus from the family Calliphoridae, which includes blow flies and bottle flies. Calliphora flies are one of the most entomologically important fly species because of their relative time of arrival and colonization to animal carcasses. Until now there are only three complete mitochondrial genome recorded from the genus. In this study we added a new complete mitochondrial genome record from the species Calliphora lata. Although genome structures and gene orientations of the four Calliphora flies mitochondrial genome are identical. The size and nucleotide composition of the genomes are slightly different.

Drosophila suzukii (Matsumura), commonly called the Spotted Wing Drosophila (SWD), originally from South Asia, is a major pest in orchards. Recently, chemical control was applied to prevent the SWD in cultivated fields, but it was not much effective because D. suzukii tended to avoid place where insecticide was treated, to reproduce in wild field, and then to recolonize neighboring cultivated fields. In contrast to chemical control, biological control can reduce these problems by using natural enemies who attack the D. suzukii. Some species of genus Asobara (Hymenoptera; Braconidae; Alysiinae) are parasitoids of Drosophila, and, In South Korea, there are 3 species recorded parasitizing in D. suzukii. In this study, we used morphological characters and mitochondrial COI gene to identify Asobara species. As a result, one species was found as a possible parasitoid of D. suzukii.

The genus Aphidius is the most commercially available species in greenhouse control, especially for aphids. Although twelve species were recorded in Korea, we collected 7 species including 2 unrecorded and 1 unidentified species. We aligned 16 sequences, except for duplicates, from 31 individuals of 14 species including GenBank sequences, and made phylogenetic tree based on Neighbor-Joining method. Surprisingly, 3 species were clustered into a clade and some species was not monophyletic. In this study, we reconstructed phylogenetic tree of Korean Aphidius species using COI marker, and compared them with GenBank sequences. In addition, we try to experiment a new marker and check its ability of discrimination.

One species of the genus Ephedrus Haliday, 1833 (Hymenoptera: Braconidae: Aphidinae) is recognized in this study. The Ephedrus genus is known to 72 species worldwide. Of these, 16 species were reported in Korea. This genus is parasitic to groups of aphids and their relatives. Additionally, this genus is very similar to genus Toxares Haliday, 1840 in the morphology, but there is no difficulty in sympathy because there are certain morphological differences. In this study, Ephedrus blattnyi Starý, 1973 is recognized for the first time in South Korea. Information on this species has not been widely known yet. Descriptions, diagnosis distribution and illustration are provided.

Origin of the red imported fire ant (RIFA, Solenopsis invicta) is South America, a tropical climate region. It has settled in the invasive area, causing various problems. In recent, colonies of Solenopsis invicta were discovered in Busan in 2017, and then in Busan, Pyongtaek, Incheon, Daegu and Ansan in 2018. This study aims to validate 31 microsatlite loci referred from previous study for tracing origin of Solenopsis invicta colonies. We tried to test previously developed microsatellite loci so that we establish the protocol for molecular epidemiology of RIFA. We applied 31 microsatellite loci to DNA samples from the RIFA colonies found in Texas and Florida, USA. We will present 31 markers were divided into three injections and the amplification rate for each marker was shown as follows.

In the 21st century, the risk of the exotic pest being invaded is increasing due to the rise of trading activities. Oriental fruit fly, Bactrocera dorsalis (Diptera: Tephritidae), has been considered to very destructive invader as it is highly intrusive and has a wide host range. In this study, 2,299 samples were collected from 88 locations in 12 countries by quarantine and investigation. Among them, 608 individuals (including B. correcta) were used for COI DNA barcoding analysis based on Neighbor Joining method with P-distance model. Population genetics analysis was conducted for 510 individuals selected from 47 locations of 12 Southeast Asian countries using 15 microsatellite loci. The barcoding analysis resulted in that any clade was not clustered according to a geographical isolation but indicated geographically mixed populations. Population genetics analysis showed shared genetic structure between neighboring countries across borders. Genetic structure of most Korean quarantine groups was more similar to that of Taiwan, China and Thailand in the order of appearance than other countries.

The morphological identification of alate aphids needs reliable and appropriate methods such as morphometrics. In previous studies, the seven species which aphids have been known as migrating to potato had been separated into genus levels using specific points of forewing vein (Landmarks). Adding the number of secondary sensillum of antennae, setae of abdomen, etc., we experimented to separate four species of genus Aphis (Aphis craccivora, Aphis fabae, Aphis gossypii, Aphis spiraecola). After taking each of 25X twenty photographies of four species, we calculated and measured coordinates of landmarks, the number of secondary sensillum of antennae, dorsal setae of 8th abdominal segment, anterior/posterior genital plate setae and cauda, and then performed the principal component analysis using PAST3 and the canonical variate analysis using MORPHO J, only with a landmarks. As results, in PCA, Aphis gossypii has been separated, but the others are overlapping. In CVA, all species of aphis has been separated.

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.

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.

Sacbrood virus (SBV) caused significant colony collapse in Korean Apis cerana. Considering that hygienic behavior in honey bees confers colony-level resistance against brood diseases, we utilized this trait for selecting A. cerana colonies. In addition, the brood survival rate was evaluated after colonies were SBV-inoculated. Over four selective generations, dead brood removal and brood survivorship in selected colonies were higher than those in the unselected colonies (P < 0.01, 99.3 vs. 89.9% for removal of pin-killed pupae; P < 0.01, 99.0 vs. 63.9% for removal of SBV-killed larvae; and P < 0.01, 70.0 vs. 9.2% for brood survivorship). Following SBV-inoculation, selected colonies showed an increase in the number of surviving pupae and adults, whereas unselected colonies collapsed mostly. Our results confirm the feasibility of selecting SBV-resistant A. cerana.

Prostaglandin E2(PGE2) is an autocrine and paracrine signal in insects and other animals. Its signal pathways in target cells are well understood in mammalian system but not in insects. Here, we assessed PGE2 signaling in hemocytes of Spodoptera exigua through knocking-down of signal component genes by RNA interference (RNAi) and knocking-out (KO) of PGE2 receptor using CRISPR-Cas9. From S. exigua transcriptome, we selected hemocyte signaling components and analyzed their functions in cellular immune responses through RNAi. KO mutant against PGE2 receptor exhibited severely hampered larval development and adult fecundity.

Insulin/IGF signaling (IIS) regulates multiple physiological processes such as larval growth, reproduction, and life span in many organisms including legume pod borer, Maruca vitrata (Lepidoptera: Crambidae). RNA interference of IIS components, insulin receptor (InR) and Forkhead Box O (FOXO), impaired larval growth and female reproduction. To further validate the physiological roles of InR and FOXO, we generated knock-out (KO) mutants using CRISPR/Cas9-mediated genome-editing technology. Both KO mutants exhibited delayed larval growth and reduced pupal and adult body sizes. In conclusion, these results demonstrated the critical role of insulin signaling (IIS) pathway to control M. vitrata growth and development.

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.

In this study, we developed 11 microsatellite markers specific to A. crataegi using NGS to investigate the genetic relationships of A. crataegi populations from South Korea to circumferential Asian countries (China, Russia, Mongolia, and Japan). Further, two mitochondrial DNA (mtDNA) gene segments (COI and CytB) were sequenced from the samples. The population- and individual-based Principal Coordinates and STRUCTURE analyses collectively suggested that the South Korean population of A. crataegi is most differentiated from the Japanese population, whereas it was closer to Mongolian and Chinese populations. These results collectively suggest that northern populations, in particular, Mongolian populations can be considered as the most genetically compatible one as donee population, when the reintroduction program is launched. †These authors contributed equally to this paper.

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.

Drosophila suzukii Matsumura (Diptera: Drosophilidae) is known as a pest that has a wide host range. This study was carried out to investigate the occurrence of D. suzukii by trapping D. suzukii in Korean strawberry greenhouse and its surrounding woodland from November, 2017 to April, 2019. As a result of trap monitoring, the greatest capture was recorded in fall and secondary peak was confirmed in summer in a lower capture. It was confirmed that there was a statistically significant difference between the occurrence among inside and outside of the greenhouse and the woodland. The strawberry greenhouse cultivation area of Korea is mainly located in the plain region, whereas most of the other host plants for D. suzukii are usually grown at the ridge of the mountain or hillside. Therefore, if considering the living environment of host plant, monitoring result that showed more capture in woodland than strawberry greenhouse would be explained.

We investigated developmental characteristic of life cycle of the lawn cutworm, Spodoptera depravata (Lepidoptera: Noctuidae), which is one of the most economically damaging pests to grasses. For larval culture the turfgrass, Zoysia japonica (Poales: Poaceae), was provided at 25℃ and 60 ± 5% humidity. Developmental period of each stage was as follows: 4.11 ± 0.19 days for eggs, 25.17 ± 3.02 for larvae, 8.80 ± 0.28 for pupae and 7.57 ± 0.95 for adults. Larvae was grown to 7th instar, unlike pre-existing data, which indicated the final age as the 6th instar. Body length of larvae changed on average from 2.772 mm immediately after hatching to 2.3 cm at 7th instar and head capsule width changed on average from 0.423 mm at 1st instar to 2.82 mm at 7th instar. Pre-period of eggs deposition was 2 days, oviposition period was 4.6 days, average fecundity per female was about 419 eggs, and hatching rate was about 83%.

A specific serotonin receptor (Se-5HTR) has been identified in the beet armyworm, Spodoptera exigua and classified into 5-HT7 type. Se-5HTR expression was up-regulated in hemocytes and fat body in response to immune challenge. As being a GPCR, this receptor is presumably coupled with intracellular trimeric Gαs protein activating cAMP-dependent protein kinase (PKA) pathway to regulate several cellular functions. RNA interference (RNAi) of Se-5HTR as well as its downstream signal proteins exhibited significant suppression in cellular immune responses including nodulation and phagocytosis. Application of inhibitors to the signaling cascade suppressed the immune responses as well. To validate the Se-5HTR involvement in mediating cellular immunity, 5-HTR knock-out mutants were developed using CRISPR-Cas9 technique and suffered significant developmental anomalies.