Insect growth regulators (IGRs) are insecticides that disrupt the normal development of target insects. Among the IGR insecticides, juvenile hormone (JH)-related IGRs are of particular interest because they stimulate or interfere with the formation of JH receptor complex. In the precious studies, novel JH-related IGRs with JH agonist (JHA) and antagonist (JHAN) activity were identified by using yeast two-hybrid system transformed with the Aedes aegypti JH receptor complex. In this study, the transcriptomic responses of Aedes albopictus were investigated upon JHA and JHAN, respectively. These results will provide important information about understanding of impact of JH-related IGRs in transcription level.

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.

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.

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.

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.

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.

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.

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.

The small brown planthopper (SBPH), Laodelphax striatellus, which transmits rice stripe virus (RSV) is one of thenotorious pests of rice plants. To investigate the factors of vector insect responding to virus introduction, the total RNAof RSV-viruliferous SBPHs was subjected to RNA-Seq to analyze the SBPH-RSV interactome. Based on the transcriptomicchanges of RSV carrying SBPHs, eight cDNA sequences possibly related to RSV replication in SBPH were selected,and dsRNAs targeting those sequences were synthesized and applied to SBPH. Three of those dsRNAs showed over 90%of substantial RSV suppression efficiency in SBPH, that demonstrated a potential of RNAi-based virus-vector managementprogram.

Secondary metabolites isolated from Actinomycete have been studied to find potent pest control agents as their insecticidal and growth inhibitory activities. In order to investigate novel insecticidal compounds, second metabolites from 363 Actinomycete isolates were evaluated for their insect growth regulatory activities. Among them, ethyl acetate extracts from ten Streptomyces spp. showed high level of Juvenile hormone antagonist activity. In addition, their insecticidal activities were tested against larvae of Aedes albopictus, Plutella xylostella and Laodelphax striatellus. These results suggested that secondary metabolites from Streptomyces spp. could be used for development of novel IGR-based insecticides.

Plant essential oils have been reported to show repellent, insecticidal and growth-reducing effect on many insect herbivores. Recently, it has been reported that plants synthesize secondary metabolites regulating insect juvenile hormone (JH) receptor complex as a part of their defense mechanisms. We tested 180 plant essential oil compounds using a yeast two-hybrid system transformed with the Aedes aegypti JH receptor as a reporter system. Among them, we identified 17 compounds that showing high JH agonist (JHA) or antagonist (JHAN) activities. They were grouped into 4 groups by their structural similarity and their insecticidal activity was tested against larvae of Aedes albopictus, Plutella xylostella, Plodia interpunctella and Laodelphax striatellus. These results could provide insights on the plant-insect coevolution and may be useful for the development of insect specific and safe pesticides.

RNA interference (RNAi) is an universal gene-knockdown mechanism in eukaryotic organisms including insects. RNAi has been considered as an alternative strategy to control agricultural pests whereby double-stranded RNA triggers a potent and specific inhibition of its homologous mRNA. Bacillus thuringiensis (Bt) is a spore-forming bacterium that produces a copious amount of crystal proteins δ-endotoxins under the control of sporulation-dependent promoter. In order to develop dsRNA mass-production platform utilizing recombinant Bt, the pHT1K-EGFP which expresses dsRNA against EGFP under the control of Cyt1-Aa sporulation-dependent promoter was constructed and the expression level of transgene (EGFP) was confirmed by qPCR analysis. These results suggested that Bt’s potential of becoming a new platform in dsRNA production.

Insect growth regulators (IGRs) are insecticides that disrupt the normal development of target insects by inducing symptoms such as premature molting or supernumerary larval stages. Juvenile hormone (JH)-based IGRs are of particular interest because they simulate or interfere with the formation of juvenile hormone receptor complex. Insect larvae generally have been poorly susceptible to JH agonists (JHAs) because endogenous JH titer is already high in the larval stage. In contrast, JH antagonists (JHANs) might be more effective for control of target pests in larval stages through interference in the status quo action of JHs. In this study, we tested 2,354 chemical compounds using the yeast-two hybrid system to identify JHANs. Among them, K265674 sample was selected and its insecticidal activity was evaluated against Aedes albopictus larvae. These results may be useful for development of novel IGR insecticide.

Entomopathogenic fungi have been widely studied for their potential as the effective biological control agents. They produce variety of secondary metabolites with insecticidal activities, and it is reasonable to assume that entomopathogenic fungi might produce secondary metabolites modulating juvenile hormone for their survival against defense mechanisms of host insect. In this study, acetone extract of the Lecanicillium spp. cultured on unpolished rice medium showed juvenile hormone antagonist (JHAN) activity in the yeast-two hybrid β-galactosidase assay and high insecticidal activity against Aedes albopictus and Plutella xylostella. In addition, to compare bioactivities of secondary metabolites from solid and liquid culture, the Lecaniciilium spp. strain cultured on unpolished rice medium or PDB medium were serially extracted with acetone and ethyl acetate respectively. Both extracts showed JHAN activity and high insecticidal activity against A.albopictus. Theses results suggested that secondary metabolites of entomopathogenic fungi could be useful for development of novel IGR insecticides.

Rice stripe virus (RSV) is one of the serious plant pathogenic viruses for rice transmitted by small brown planthopper (SBPH), Laodalphax striatellus. RNA interference (RNAi) was recently suggested as a promising strategy for controlling insect pests, including those that serve as important vectors for plant pathogens. To disturb transmission of the RSV in SBPH using RNAi pathway, we chose nine genes highly expressed in RSV-viruliferous SBPH by transcriptome sequencing. These SBPH-derived dsRNAs were applied to the insects indirectly through xylem of rice leaves by irrigation. qPCR result demonstrated that five out of nine SBPH-derived dsRNAs successfully reduced the replication of RSV in viruliferous SBPH in dose-dependent manner, suggesting that these five dsRNAs could suppress replication of RSV and provide a new tool for RSV control strategy.

The small brown planthopper (SBPH), Laodelphax striatellus, is one of the most serious pest insects of rice plants because it can transmit the rice stripe virus (RSV) which often causes significant reduction of yield in the field. Buprofezine is an effective insect growth regular (IGR) pesticide to control planthoppers, however, since the use of buprofezine for more than a decade, it has caused a certain resistance of SBPH. To survey the responses of SBPH to buprofezine, we exposed 4th instar SBPH to 200 ppm buprofezine by dipping method, and extracted total RNA for RNA-seq by Illumina platform. The quality filtered raw reads of cDNA obtained from experimental and control SBPH were subjected to Bowtie2 followed by eXpress computer program to compare the differential gene expression which will be important information for pest control methods using RNAi.

Bacillus thuringiensis (Bt) produces a variety of insecticidal crystal proteins and widely used as one of the most successful biological control agents. Recently, studies that introduce cry genes into crops to create pest resistance have made much progress, and the total area of land planted with Bt crops has increased substantially. In this study, pest resistance of 8 transgenic Bt rice events with a synthetic cry1Ac gene linked to rice rbcS-tp sequence were assessed under laboratory conditions. Bioassays were performed against Cnaphalocrocis medinalis, which is a significant pest of rice in Asia. C. medinalis larvae were shown to be susceptible to all eight events, even though there were differences between the causes of death. The results differed between developmental stages of the larvae, despite the fact that all 8 events led to high mortalities. These results may be a significant foundation for the evaluation of improved transgenic Bt rice.

Insect growth regulators (IGRs) are attractive pest control agents due to their high target specificity and relative safety to the environment. Recently, plants have been shown to synthesize IGRs that affect the insect juvenile hormone (JH) as a part of their defense mechanisms. We identified several JH agonists (JHAs) and antagonists (JHANs) from plant essential oil compounds using a yeast two-hybrid system transformed with the Aedes aegypti JH receptor as a reporter system. They showed high mosquitocidal activities with relatively low LC50 values and caused retardation of ovarian development in female mosquitoes. While the JHAs increased the expression of JH-induced gene, the JHANs caused reduction in the expression of the same gene. The compounds identified in this study could provide insights on the plant-insect interactions and may be useful for the development of novel IGR insecticides.

Entomopathogenic fungi have been widely studied for their potential as the effective biological control agents. They produce variety of secondary metabolites with insecticidal activities, and it is reasonable to assume that entomopathogenic fungi might produce secondary metabolites modulating juvenile hormone for their survival against defense mechanisms of host insect. In this study, Beauveria spp. and Lecanicillium spp. strains cultured on unpolished rice medium were extracted with acetone. Both extracts showed juvenile hormone antagonist (JHAN) activities in the yeast-two hybrid β-galactosidase assay. In addition, they showed high insecticidal activities against Aedes albopictus, Plutella xylostella, and Ostrinia furnacalis. These results suggested that secondary metabolites of entomopathogenic fungi could be useful for development of novel IGR insecticides.

Streptomyces is the largest genus of Actinobacteria that forms fungus-like branched networks of hyphae. Streptomyces has been clinically important because they produce various secondary metabolites with antibacterial, antifungal, and nematocidal activities. In order to explore novel insecticidal compounds, extracts from 363 strains of Actinobacteria were screened for their juvenoid and anti-juvenoid activities using yeast-two hybrid system. Among them, extract of Streptomyces spp. showed high anti-juvenoid activity. This extract also showed high level of insecticidal activities against larvae of Aedes albopictus, Laodelphax striatellus, and Ostrinia furnacalis. These results suggested that the secondary metabolites of Streptomyces could be natural sources of novel insecticidal compounds.