배초향은 항동맥경화나 항박테리아의 특성을 가지는 한약재에 널리 사용되는 영년생 약용식물이다. 연구의 목적은 수경재배에서 배양액의 종류와 PPFD값에 따른 배초향의 생장 및 항산화 물질의 변화를 조사하는 것이다. 배초향은 주야간 16:8 시간의 일장조건에서 150과 200 μmol·m-2·s-1 PPFD 조건과 일본원시(HES), 서울시립대(UOS), 유럽채소연구소(EVR), 오오츠카 배양액(OTS)을 이용하여 6주간 재배하였다. OTS 배양액조 건에서 자란 배초향의 지상부 및 지하부 건물중은 다른 배양액 처리구와 비교하여 유의적으로 높았다. 배초향의 틸리아닌 함량은 OTS 처리에서 가장 높았으며 다음으로 EVR, HES, UOS 순서로 낮아졌다. 총 아카세틴의 함량은 EVR처리에서 가장 높았으나 OTS처리와는 유의적 차이를 보이지 않았다. 또한 200 μmol·m-2·s-1 PPFD 조건에서 자란 배초향은 PPFD 150처리구와 비교하여 유의적으로 생체중과 건물중이 증가하였으며 기능성 물질은 틸리아닌과 아카세틴의 함량도 높았다. 본 연구는 수경재배 방식을 이용하여 식물공장에서 배초향을 재배할 경우 200 μmol·m-2·s-1 PPFD 조건과 OTS 배양액 조건에서 경제적인 광원조건으로 최적 바이오매스 생산량과 틸라아닌과 아카세틴의 함량을 증가시킬 수 있을 것으로 제안한다.

As the fruit and vegetable beverage market grows, a variety of foods are continually increasing. Therefore, when ingredients other than those indicated are added to achieve economic benefits, cases of adulteration and falsification concurrently follow. Among these, blueberry as expensive fruits, is one of the target for adulteration in juice production. This study was conducted based on the reports regarding the forgery of blueberry juice and grape juice; 32 kinds of blueberry juices, which are sold on the market, were collected and their metabolomics analysis was performed to screen out possible discriminants for blueberry juice adulteration. Metabolomes were extracted with 80% methanol and analyzed through LC-MS/MS followed by data processing with multivariate statistical analysis. Based on OPLS (orthogonal partial least squared) model, four metabolites were screened as significant discriminants among 209 metabolites found in blueberry juice and anthocyanin compounds occupied a main groups for discrimination. Marvidin-3-O-glucoside and cyanidin-3-O-rutinoside were identified as significant indicators for the existence of blueberry compared to grape juice which is main adulterates in blueberry juices. These candidates were assessed for monitoring commercial blueberry juices, which were proved as useful determinant for adulteration.

Little consideration has been given to the insect epizootiology in pest management. High production cost of fungalinsecticides might be a technical barrier. Application of the fungal insecticides to unfavorable conditions might result inlower performance in pest management. Herein this work, we suggest ecological biocontrol considering long-term colonizationin nature rather than quick pest control. Fungal biopesticides should be frequently combined with chemical pesticides infields, particualrly spatial combination strategy rather than temporal combination. Additionally other important technicalaspects need to be strongly considered, such as economic downstream process, effective control and environmentally safe,so finally proposing e-biopesticide. Lastly R&D system needs to be clearly and efficiently organized for high performance.Week-based summary and reporting system, weekly report and project file are main system in our laboratory which hasour lab work more effectively and efficiently, consequently resulting in tech-transfer to an industry and re-investment.

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.

Bee venom contains a variety of peptide constituents, including low-molecular-weight protease inhibitors. While the putativelow-molecular-weight serine protease inhibitor Api m 6 containing a trypsin inhibitor-like cysteine-rich domain was identifiedfrom honeybee (Apis mellifera) venom, no anti-fibrinolytic or anti-microbial roles for this inhibitor have been elucidated.In this study, we identified an Asiatic honeybee (A. cerana) venom serine protease inhibitor (AcVSPI) that was shownto act as a microbial serine protease inhibitor and plasmin inhibitor. AcVSPI was found to consist of a trypsin inhibitor-likedomain that displays ten cysteine residues. Interestingly, the AcVSPI peptide sequence exhibited high similarity to the putativelow-molecular-weight serine protease inhibitor Api m 6, which suggests that AcVSPI is an allergen Api m 6-like peptide.Recombinant AcVSPI was expressed in baculovirus-infected insect cells, and it demonstrated inhibitory activity against trypsin,but not chymotrypsin. Additionally, AcVSPI has inhibitory effects against plasmin and microbial serine proteases; however,it does not have any detectable inhibitory effects on thrombin or elastase. Consistent with these inhibitory effects, AcVSPIinhibited the plasmin-mediated degradation of fibrin to fibrin degradation products. AcVSPI also bound to bacterial andfungal surfaces and exhibited anti-microbial activity against fungi as well as gram-positive and gram-negative bacteria. Thesefindings demonstrate the anti-fibrinolytic and anti-microbial roles of AcVSPI as a serine protease inhibitor.

Major royal jelly proteins (MRJPs) are important protein components of bee royal jelly (RJ) and exhibit various biologicaland pharmacological activities. The antimicrobial activities of royalisins and the jelleines contained within MRJP 1 andMRJP 2 in RJ have been elucidated. However, the antimicrobial effects of other bee RJ MRJPs remain largely unknown.In this study, we demonstrated that the Asiatic honeybee (Apis cerana) MRJP 4 (AcMRJP4) exhibits antimicrobial activitiesagainst bacteria, fungi, and yeast. Recombinant AcMRJP4 was expressed as a 63-kDa protein in baculovirus-infected insectcells. However, some of the recombinant AcMRJP4 proteins were cleaved into two fragments (i.e., 48-kDa (AcMRJP4-48)and 15-kDa (AcMRJP4-15) proteins) by the proteolytic cleavage of the C-terminus of the recombinant AcMRJP4. Interestingly,AcMRJP4, AcMRJP4-48, and AcMRJP4-15 exhibited antimicrobial activities, with AcMRJP4-15 exhibiting the highestantimicrobial activity, followed by AcMRJP4. AcMRJP4-15, which is a hydrophilic peptide with 88 amino acid residuesthat contains a high content of Asn and positively charged amino acids, induced structural damage in the cell walls ofthe assayed bacteria, fungi, and yeast. Altogether, our data demonstrated that AcMRJP4 functions as an antimicrobial agent.

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.

The hard tick, Haemaphysalis longicornis (Ixodida: Ixodidae) is one of the vectors of severe fever with thrombocytopeniasyndrome virus (SFTSV) causing in humans, however little consideration was given to the biological control of the pest.We evaluated three bioassay methods, such as spray, dipping and dropping. As a result, the dipping method was themost efficient way to measure the fungal virulence. Finally, we constructed a Haemaphysalis longicornis pathogenic fungallibrary to further facilitate the resources to be used as potential biological control agents. A conidial suspension of 1×107conidia/ml was exposed to the ticks by dipping. We confirmed that some isolates were highly virulent to the ticks. Thisresults indicated that some fungi could be used to effectively control the Haemaphysalis longicornis.

Entomopathogenic fungi have been widely studied for their potential as the effective biological control agents. Theyproduce variety of secondary metabolites with insecticidal activities, and it is reasonable to assume that entomopathogenicfungi might produce secondary metabolites modulating juvenile hormone for their survival against defense mechanismsof host insect. In this study, Acetone extracts of 189 entomopathogenic fungi cultured on unpolished rice medium werescreened for their juvenile hormone antagonist (JHAN) activities using the yeast-two hybrid system. Among them, 14extracts showed high level of JHAN activities and their insecticidal activities against Aedes albopictus were investigated.

The small brown planthopper (SBPH), Laodelphax striatellus, is one of the most serious pest insects of rice plants.Buprofezin has been used to control SBPH for more than a decade, however, the occurrence of buprofezin resistant SBPHwas reported recently. To develop an alternative pest control an alternative pest control strategy, RNA-seq of buprofezin-treatedSBPH was performed to screen the insecticidal target genes for RNA interference (RNAi). Six genes were selected fordsRNA synthesis, and applied to SBPH to assess the insecticidal efficacy. Two and three of those dsRNAs showed moderatedand substantial insecticidal activity up to 60% of mortality in one week, respectively. These results demonstrated the potentialof gene screening strategy for the development of RNAi-based pest management program.

Actinobacteria have been known to produce variety of bioactive metabolites that include antibiotics and plant growthfactor. Also, Insecticidal compounds such as avermectin and tetranectin have been reported from actinobacteria. In thisstudy, to identify novel insect growth regulator (IGR)-based insecticidal compounds, 1,283 actinobacteria isolates weretested for their juvenile hormone antagonist (JHAN) activity using the yeast-two hybrid β-galactosidase assay. Amongthem, 25 isolates showed high level of JHAN activity and insecticidal activity against 3rd larvae of diamondback moth,Plutella xylostella. These results suggested that actinobacteria could be unparalleled source of novel IGR-based compounds.

Widespread use of chemical insecticides has resulted in development of insect resistance and natural products withbiological activities could become an attractive alternative to control insect pests. In order to find more effective insecticidesfor controlling mosquito, various mosquitocidal compounds are studied. Recently, juvenile hormone antagonists (JHANs)are known to be to safe and effective insecticides for control mosquito. In order to identify novel insecticidal compoundswith JHAN activity, several chalcones were surveyed on their JHAN activities and larvicidal activities against Aedes albopictuslarvae. Among them, 2’-hydroxychalcone and cardamonin showed high level of JHAN and mosquito larvicidal activities.These results suggested that chalcones with JHAN activity could be useful for control of mosquito larvae.

Recently, it has been reported that plants produce variety of phytochemicals against insect pests as a part of their defencemechanism. In this study, to identify novel compounds that disrupt juvenile hormone (JH) receptor complex, plant essentialoil compounds were tested using in vitro yeast two-hybrid system transformed with the Aedes aegypti JH receptor complex.Among the plant essential oil compounds, farnesol interfered with pyriproxyfen-mediated binding of A. aegypti Met-FISC.In contrast, farnesyl acetate induced the binding of A. aegypti Met-FISC. Also, both compounds caused retardation ofovarian development and change of transcription level in female A. albopictus. These results could provide insights onthe plant-insect coevolution and may be useful for the development of insect specific and safe pesticides.

Streptomyces species have been studied to find potent pest control agents as an alternatives to chemical insecticides.Previously, one of the ethyl acetate extract of Streptomyces isolates cultured on unpolished rice medium showed highlevel of juvenile hormone antagonist and larvicidal activities against pest insects including Aedes albopictus, Plutella xylostellaand Laodelphax striatellus. It has been known that the biosynthesis of secondary metabolites of Streptomyces could beinfluenced by a variety environments such as nutritional composition and growth conditions. In this study, to optimizeculture conditions for stable production of insecticidal compounds from this isolate, binding assay and bioassay-guidedmonitoring were conducted using various culture conditions.

RNA interference (RNAi) has been considered as an alternative strategy to control agricultural pests whereby double-strandedRNA triggers a potent and specific inhibition of its homologous mRNA. Since small double-stranded RNAs are requiredfor various RNAi applications, there is a need for cost-effective methods for producing large quantities of high-qualitydsRNA. Bacillus thuringiensis produces much insecticidal proteins with expression of their encoding genes being drivenby sporulation-dependent promoters. To develop dsRNA mass-production platform utilizing Bt, the pHT1K-EGFP plasmidvector which has cyt1Aa sporulation-dependent promoter was constructed. The transcriptional level of target gene (EGFP)is higher 113 times than Bt reference gene (ssPE). It was applied to protect honeybee from Sacbrood virus, so targetgene was replaced to SBV-vp1. By ingestion of Bt-derived dsRNA to honeybee shows positive effect on SBV suppression.

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.