Beauveria bassiana JEF-007 with strong virulence against Riptortus pedestiris was selected for the Agrobacerium tumefaciens-mediated transformation(AtMT). AtMT generated two transformants, B1-06 and C1-49, showed significantly reduced virulence against R. pedestris. To identify the virulence-related genes, thermal asymmetric interlaced(TAIL) PCR and flanking region analysis were performed. From the analysis, two genes, Complex I intermediated-associated protein 30(CIA30) and Autophagy protein 22(Atg22), possibly related virulence in B. bassiana JEF-007. For the analysis of two putative virulence-related genes in JEF-007, hairpin RNA interference (hpRNAi) is under consideration. This work can provide the functional roles of the virulence-related genes in B. bassiana JEF-007.

Chemical pesticides have been used to control persimmon pests, however the overuse of the pesticides caused insect resistance. Herein we investigate the potential of eco-friendly organic pesticides (EFOP) on the control persimmon pests, Stathmopoda masinissa and Riptortus pedestris. Eleven commercially available EFOP were sprayed on the target insects in laboratory conditions. The chemical pesticide, buprofezin+dinotefuran wettable powder served as a positive control. In the bioassay against persimmon fruit moth, the mixture of Chinese scholar tree, goosefoot and subtripinnata extracts (EFOP-2) treatment showed the highest control efficacy: 27.7% (5 days) of survival rates. In the bioassay against bean bugs, the EFOP-2 showed the highest control efficacy: 20.0% (5 days) of survival rates. These results suggest that the mixture of three plants extracts (EFOP-2) has high and multiple potential in the management of the persimmon pests.

A viral genome was assembled de novo from next-generation sequencing (NGS) data from bean bugs, Riptortus pedestris, infected with an entomopathogenic fungus, Beauveria bassiana (Bb), and was further confirmed via the RACE method. Based on the phylogenetic analysis of the RdRp sequences, RiPV-1 was clustered in the unassigned insect RNA viruses with two other viruses, APV and KFV. These three viruses were suggested to constitute a new group of insect RNA viruses. Interestingly, RiPV-1 replication was increased dramatically in bean bugs 2 to 6 days after fungal infection. In conclusion, a novel insect RNA virus was found by NGS data assembly. This virus can provide further insight into the interaction between virus, fungus and the host.

Currently, only limited number of mitochondrial genomes (mitogenome) is available from Odonata. In order to extend current mitogenome data for comparative biology and phylogeny we sequenced complete mitogenomes of two endangered dragonfly species, Libellula angelina and Nannophya pygmaea (Ododana: Libellulidae). The whole genomes were 15,233 bp in L. angelina and 15,112 bp in N. pygmaea and included a typical set of genes (13 protein-coding genes, two rRNA genes, and 22 tRNA genes) and one major non-coding A+T-rich region. The arrangement of the genomes was identical to typical one found in insects. Phylogenetic reconstruction using concatenated sequences of 13 PCGs and two rRNAs of Odonata (17 species in eight families in three suborders) using both Bayesian Inference (BI) and Maximum Likelihood (ML) methods have shown a strong support for monophyletic Zygoptera (BI, BPP = 1 and ML, 100%). Currently, further scrutinized analysis is under progress.

The number of reported mitochondrial genomes (mitogenomes) from the monotypic Lasiocampoidea has been limited until recently. In this study, we sequenced the complete mitogenome of the lappet moth, Kunugia undans (Lepidoptera: Lasiocampidae), and compared it to those of other lasiocampid species and macroheteroceran superfamilies (59 species in six superfamilies). The 15,570-bp long K. undans genome had the typical set of genes found in animal mitogenomes, with the exception of one additional trnR that are located between trnA and trnN loci. Considering that the two trnR copies are located in tandem with proper secondary structures and identical anticodons, a gene duplication event might be responsible for the presence of the two tRNAs. In summary, the general mitogenome characteristics of Lasiocampoidea did not differ greatly from the remaining macroheteroceran superfamilies, but it did exhibit some unique features.

The spotted-wing drosophila (SWD), Drosophila suzukii (Diptera: Drosophilidae), is an economically damaging pest that feeds on most thin-skinned fruits. In this study, we sequenced portions of the mitochondrial (mt) COI and ND4 genes from a total of 195 individuals collected mainly from Korea. A total of 139 haplotypes were obtained from the concatenated COI and ND4 sequences. A dataset combining GenBank sequences with our own data identified a total of 94 worldwide COI haplotypes with a maximum sequence divergence of 5.433% (32 bp). A rough estimate of genetic diversity in each country showed higher diversity in ancestral distributional ranges, but the invasion over Asian countries seems to have been substantial because haplotype diversity was only 2.35-3.97-fold lower in the USA, Canada, and Italy than that in the populations ancestral ranges.

Caloptilia Hübner, 1825 is one of the largest genera in Gracillariidae. 314 species have been known around the world. To date, 19 species were known from Korea. In the present study, one species, C. acericola (Kumata, 1966), is newly recognized from South Korea. Caloptilia acericola was first described from Japan and it also distribute Russian Far East. The name of this species was derived from the genus name (Acer, Aceraceae) of its host plant. We redescribed with illustrations of adults and genitalic structures to distinguish the species from the congeneric species in South Korea.

Alternative control agents, hopefully overcoming the present issues and problems in agrochemicals, should be considered and finally applied to the vector controls. Entomopathogenic fungi can used as one of the possible novel control agents and great considerations are given to the control of soil- or water-dwelling stages of vectors, such as mosquitoes and ticks. In our research group, the entomopathogenic fungal library has been constructed using the mealworm-based isolation system, which showed a variety of opportunities of their use in vector control. Important key production technologies including granular formulation have been developed to increase their industrialization. Some entomopathogenic fungal isolates showed high biological performance in the control of mosquitoes and ticks in field stands. R&D of down-stream process should be seriously considered as the key step.

Currently, phylogenetic relationships of Macroheterocera with the inclusion of Bombycoidea have been reconstructed using molecular data, but generated several hypotheses on familial relationships within Bombycoidea and among-superfamilial relationships within Macroheterocera. Thus, in this study, mitogenome-based superfamilial relationships of Macroheterocera and familial relationships among macroheteroceran superfamily, Bombycoidea, were tested with different algorithms and partition schemes. Additionally, two unrecorded species of Bombycoidea (Saturnia jonasii and Kentrochrysalis streckeri) detected in Korea using morphological and molecular data are presented.

Zika virus has become a problem in the world recently. Zika virus is spread by several different species of Aedes mosquitoes, Aedes albopictus is one of them. The Asian tiger mosquito, Ae. albopictus, is already inhabit the Korea. Therefore, control of this mosquito is very important and urgently needed. Environmental friendly control methods to a change in the recognition of people have been desired. Essential oils and plant extracts may use an alternative method to control the mosquitoes. This study describes about the insecticidal and repellent against Ae. albopictus using plant materials.

A blast search using SPODOBASE gave 10 chymotrypsin (CHY) candidate genes with complete open reading frames (ORFs) of Spodoptera exigua. Among these candidates, Se-CHY2 was chosen and further analyzed in expression and RNAi. Se-CHY2 was expressed only in larval stage and especially in midgut. Its RNAi was successfully performed by double injection of specific double-stranded RNA (301 bp). qPCR showed a decrease of Se-CHY2 expression level by ~50% at 48 h and ~83% at 72 h. RNAi resulted in significant larval mortality. All treated larvae died at 96 h after dsRNA injection. In contrast, control dsRNA treatment did not give any mortality. Enzyme activity of gut juice was measured with N-succinyl-alanine-alanine-prolin-phenylalanine-p-nitroanilide substrate and significantly decreased in larvae treated with dsRNA. These results indicate that Se-CHY2 is a chymotrypsin that is secreted into midgut lumen and plays a role in nutrient digestion.

Phospholipase A2 (PLA2) catalyzes the committed step for eicosanoid biosynthesis. Two cellular PLA2 genes (iPLA2 and cPLA2) have been identified in Spodoptera exigua. PLA2 activity varied among developmental stages and different tissues. There was a positive correlation between PLA2 activities and their expression levels. Interestingly, there was significant inhibition by sPLA2 inhibitor, suggesting a presence of sPLA2 in S. exigua. An entomopathogenic bacterium, Xenorhabdus hominickii, infection suppressed expression of PLA2 genes and its extract inhibited enzyme activities. Oxindole of a bacterial metabolite of X. hominikii inhibited PLA2 activity and expression.

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.

Many species of braconids play an important role in controlling the populations of insects in agricultural and natural ecosystems, which parasitize pests. Nevertheless, the family Braconidae was scarcely research and DNA barcoding of Braconidae has not been studied in Korea. DNA barcoding can rapidly perform species identification, and classify difficult species in morphological identification. Because most braconids are very small and cryptic in morphology, DNA barcoding is useful in Braconidae taxonomy. In this study, genomic DNA of 280 individual samples were extracted, and, among them, 208 samples were obtained COI (cytochrome c oxidase I) barcode sequences. As a consequence of blast, 48 samples were identified in genus-level, and 51 sample were identified in species-level. Among them, 4 species were revealed as unrecorded in Korea.

Cycloamyloses (CA) from high-amylose rice starch (Dodamssal, 50.94% amylose) was produced to form a complex with anthocyanin extracted from black rice (sihtoheukmi). The effects of CA concentration (1, 5, 10 and 30%, w/v), pH (2, 4, 6, and 8), UVB irradiation time (~24h) and thermal treatment time (~24h) on the oxidant capacity of the complex were investigated. Anthocyanin consisting of 95% cyanidin-3-glucoside (C3G) was extracted from the black rice by suspending in 60%(v/v) ethanol containing 0.2%(v/v) hydrochloric acid for 90 min at 50°C. Cycloamyloses (CA) produced by 4-α-glucanotransferase (4αGTase) formed complexes with C3G by shaking the mixture for 48 hours at 30°C. Antioxidant activities (radical scavenging capacity) were determined by using ABTS [2,2'-azinobis-(3-ethylbenzothiazoline- 6-sulfonic acid)]. The antioxidant activities of the complexes increased with pH increasing. Upon UVB irradiation and heat treatment, the antioxidant activities were more retained for the complexes incorporating 10% CA concentration or above compared to the C3G control. Degradation rate constant (Kd) and half-life (T1/2) were obtained from degradation data of CA complex with cyanidin-3-glucoside. The effect of CA complex formation with C3G is significantly observed at pH4. The results suggest that the CA complex formed with C3G indeed has an enhanced antioxidant potential toward UVB and thermal degradation.

Ionic liquids (ILs) have been used in DNA extraction/separation, DNA preservation and PCR based on their characteristic affinity to DNA. However, few studies have been performed about how DNA-IL complex forms and its mechanism which would be essential to understand the role of ILs over the range of applications. Herein, we present that the differences in the structure of the DNA- IL complex are associated with the alkyl chain length of IL. The assumption was evidenced by Atomic force microscopy, DNA specific dye staining, gel-electrophoresis and real-time electrical measurement. We observed unique electrical signals with altered duration time and amplitude when DNA- ILs complexes pass through solid-state nanopore. We examined three types of ILs (EMIM-Cl, BMIM-Cl, and OMIM-Cl) for their characteristics to form DNA-ILs complexes. The results indicated that the length of hydrophobic alkyl group in respective ILs determines the form of DNA-IL complex. In conclusion, the morphology of DNA could be modified by the incorporation with different alkyl chain length of ILs, providing their further application in biosensor such as nanopore technique for DNA sequencing or understanding protein-DNA interaction.

To increase the stability of anthocyanin from black rice, β-cyclodextrin (β-CD) formed a complex with anthocyanin, and thermal and UVB stability of the complex was investigated at various pH levels. The formation of β-CD complex with anthocyanin was confirmed by the decreased UV-Vis spectrophotometer absorbance (at 511nm) with increasing the concentration of β-CD at pH 2, 2.5, 3, 4, and 5. ABTS (2, 2’-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid)assay was conducted to measure radical scavenging capacity (RSC) of the complex at pH 2, 4, 6, and 8, after which the RSCs were recorded over the 24 h heating(95°C) and 24 h UVB irradiating periods. The RSC values of the complexes increased as the β-CD concentration increased (0.1-2%) at pH 6 and 8. Upon thermal treatment, the RSC readings of the complexes tended to decrease to a lesser extent compared to the anthocyanin control (without β-CD) at pH 2. This result suggests that the thermal stability of anthocyanin was more retained in the presence of β-CD at low pH (pH 2). However, in the case of UVB irradiation, the effect of β-CD complex on the anthocyanin RSC was not significant, though the RSC values for both the anthocyanin control and complexes trended downward as the UVB irradiation time increased at all pH ranges except for pH 8 (no downward trend). In short, this study suggests that the effect of β-CD complex on the stability of anthocyanin antioxidant capacity depends on pH and the susceptibility to the degradation process.

Although carnosic acid (CaA) is known as one of the useful polyphenolic compounds due to its antimicrobial and antioxidant activities, it is limited to use as aqueous solution because of its low solubility and unstability. The objective of this study was to investigate the capability of CA to improve the solubility of CaA by forming an inclusion complex in comparison with cyclodextrin (CD) and maltodextrin (MD). Enzymatically-produced CA was reacted with CaA in aqueous solution to form a complex using a freeze-drying. And the formation of complex between CaA and CA was identified by X-ray diffraction (XRD), differential scanning calorimeter (DSC) and field emission scanning electron microscope (FESEM). As a result of XRD and DSC analysis, disappearance of characteristics of CaA that was reacted with CA could be indicated the complex formation between CaA and CA. The formation complex of CaA with CA was also confirmed through the change in morphology of CaA and CA in the electron micrograph result. Aqueous solubility of CaA with various concentrations (1, 5, 10, 20, 30%) of CA was measured by absorbance change at 285 nm. As a result, the solubility of the CaA was significantly increased with increasing CA concentration. At 30% CA, the maximum solubility of CaA was 0.095% (w/v) in solution, which was approximately 3 times higher than that of free CaA (0.033%). The effect of inclusion complex with CA on the solubility of CaA was superior than that with CD (0.057%) and MD (0.066%). These results indicated that the effects on the solubility and formation abilities of inclusion complex were associated with host materials and its concentration rate. This study confirmed that the CA can be a viable solution to improve the aqueous solubility of CaA. Further investigation is still needed to understand the effect of inclusion complex with CA.

Various methods for the detection of E. coli O157:H7 in food have been developed in the past decades. However, current detection methods require specialized instruments and lengthy preparation time. In an effort to achieve a rapid and sensitive detection, we developed a radial chromatography (RC) biosensor integrated with gold nanoparticle (AuNP) conjugated with antibody for the detection of E. coli O157:H7. The immuno-AuNP binds to E. coli O157:H7 creating AuNP-E. coli O157:H7 complexes by specific antigen-antibody interaction. The AuNP-E. coli O157:H7 complexes can be identified clearly from free AuNP by RC based on their mobility on porous matrix. Thus, the AuNP complexed with target bacteria, E. coli O157:H7 could be discriminated from free AuNP by radial chromatography. The results showed that the developed RC biosensor is highly selective to E. coli O157:H7 over non-target bacteria, including Bacillus cereus, Staphylococcus aureus and Vibrio cholerae with a detection limit of 105 CFU/ml. When combined with a pre-concentration step using immunomagnetic beads, we could further enhance the detection limit down to 103 CFU/ml. In this study, we developed a novel method that is rapid, sensitive and applicable for qualitative and quantitative detection of E. coli O157:H7. The detection procedure is simple and the results can be easily determined by naked eyes, suggesting that this system is practical and can be applied to the field diagnosis in food industry for the detection of pathogenic bacteria.

Recently, there is a growing interest of consumers in natural products thus a large number of natural preservatives have been studied as food additives. Among those natural preservatives, rosemary extract are commonly used by the food industry to extend the shelf life of several products. Nevertheless, the incorporation of the rosemary extract in food matrix is highly limited due to the low water solubility and poor chemical stability of constituents of rosemary extract. In this context, it is needed to find a way that can improve the water solubility to incorporate rosemary extract into aqueous system like foods. Hence, in this study we added surfactants that have a relatively high hydrophilic-liphophilic balance number into rosemary extract solution to increase the water solubility of rosemary extract and then investigated the microbial activities of rosemary extract with surfactants. Tween 20, decaglycerol monooleate, decaglycerol laurate and decaglycerol myristate were used in a range 0.1~4% and rosemary-surfactant solution was prepared by dissolving rosemary in surfactant added phosphate buffer (pH 7) as much as extract can be dissolved. Water solubility of rosemary-surfactant solution were examined by observance of the UV-vis spectra. The antimicrobial effects rosemary-surfactant solution on B.subtilis were also examined by paper disc diffusion method. Rosemary extract showed poor solubility in normal phosphate buffer but its water solubility was highly increased when surfactant was added. This tendency was lasted at all types of surfactant. This is because of amphipathic property of surfactant. Antimicrobial effect to B.subtilis was observed when a relatively lower concentration of surfactants were used, however, not in high concentrations. This result might be attributed to the formation of surfactant micelles containing rosemary extract when a relatively higher concentration of surfactants were used. The information presented may be useful for the development of a new rosemary-loaded delivery systems.