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.

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.

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.

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.

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.

Varroa destructor is a devastating ectoparasitic mite which attacks Honeybee, Apis mellifera. V. destructor feeds on honeybee hemolymph, and often harbors small RNA viruses such as the deformed wing virus to transmit these viruses in the infested bee hive. To survey the genes of V. destructor, total RNA was subjected to high-throughput transcriptome sequencing to construct in silico cDNA library by using the Illumina HiSeq 2000 platform. Total of 2×107,748,792 paired-end short reads were obtained and quality filtered reads were subjected to Trinity de novo assembler followed by TransDecoder, and CD-HIT program to make a V. destructor reference cDNA library containing 28,023 of clustered contigs with protein coding capacity. These cDNA sequences will help us to understand the molecular biology of V. destructor.

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.

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.

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.

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. IGRs are categorized into three types based on their mode of action. One of the them is classified as juvenile hormone agonists (JHAs). In recent studies, we identified juvenile hormone antagonists (JHANs) from plant extracts. In order to identify a novel juvenile hormone agonists and antagonists, we tested 2,354 chemical compounds using the yeast-two hybrid system. Among them, 2 JHAs and 4 JHANs were selected and their insecticidal activities were evaluated against Aedes albopictus larvae. These results may be useful for development of novel IGR insecticides.

Brachymystax lenok tsinlingensis (family Salmonidae), cold freshwater fish, is endemic to Asia. This species is currently distributed throughout Russia, Mongolia, China and the Korean Peninsula. B. lenok tsinlingensis in South Korea was severely affected by anthropogenic activities such as habitat destruction, agricultural run-off and water pollution, and hence this fish has recently been dramatically decreased in its population sizes and become now critically endangered. To recover the number of individuals of B. lenok tsinlingensis, stocking or translocation programs have been conducted continuously by local governments since 1970s. However, these programs made little effort to clarify populations that may have originated from stocked, translocated or introduced fish. An understanding of genetic characteristics of endangered populations is critical to develop effective conservation and restoration plans especially because genetic diversity ensues their future fate. Therefore, we assessed the “conservation status” of this species by estimating the level of genetic diversity and genetic structure among ten geographic populations including restored populations via reinforcement and supplementation. Also, we aimed to trace the genetic origins of the newly translocated population (Chiak) through a restoration practice program. Moreover, we inferred the phylogenetic relationships among Korean lenok populations as well as across the Northeast Asia. Two hundred eighteen individuals of B. lenok tsinlingensis were sampled from ten localities (Yanggu, Injae, Seorak, Bangtae and Hongcheon: North Han River basin; Pyeongchang, Chiak and Jeongseon: South Han River basin; Taebaek and Bonghwa: Nakdong River basin in South Korea). Based on mitochondrial DNA (mtDNA) control region and eight nuclear microsatellite loci, we found extremely low levels of within-population genetic diversity, which suggests small effective population sizes (Ne) within populations. For mtDNA control region, each population housed one, or at most, two haplotypes that are restricted to the respective localities, meaning that these ‘genetically unique’ lineages will be lost permanently if the local populations undergo extinction. The overall values of haplotype diversity (h) and nucleotide diversity (π) for the entire Korean population were 0.703 ± 0.024 and 0.021 ± 0.010, respectively. In the case of microsatellites, average number of alleles across the eight loci for the entire population was 9.1 and allelic richness (AR) per population ranged from 2.375 to 4.144 (mean = 3.104). The values of observed heterozygosity (HO) and expected heterozygosity (HE) were similar to each other [HO: 0.400 ~ 0.590 (mean = 0.518); HE: 0.407 ~ 0.608 (mean = 0.504)]. The inbreeding coefficient (FIS) values were generally low, ranging from 0.048 to 0.279. Consequently, the majority of the populations (except Yanggu and Pyeongchang) were not significantly deviated from Hardy-Weinberg equilibrium (HWE), suggesting random mating at these loci tested. In addition, we found that Korean lenok populations were significantly genetically isolated from each other, with private mtDNA haplotypes and microsatellite alleles, indicating limited gene flow among populations, strong effects of genetic drift due to small Ne, or a combination of both. The Mantel test of microsatellites revealed a significant correlation (r = 0.414, P = 0.04) between genetic and geographic distances for pairwise comparisons among the ten populations, while that of mtDNA showed a lack of correlation. Given the shared identical mtDNA haplotype and similar microsatellite allelic distributions between Chiak and Hongcheon populations, we suggest that the restored (introduced) Chiak population would be inferred to be genetically originated from Hongcheon population. Phylogenetic relationships among Northeast Asian populations showed that South Korean lineages have more recently diverged from China (Yellow River), than between North Korea and Russia. Although the phylogenetic relationship would be expected to be associated with geography, South-North Korea and China populations with a similar latitude was more phylogenetically closely related. These findings may suggest a possible scenario for the historical movements of B. lenok tsinlingensis in Northeast Asia during Last Glacial Maximum (LGM). It would be supported by the line of evidence that most lenok populations migrated to southward from Northern Asia such as Russia and Mongolia during LGM because the Korean Peninsula was landlocked as inland epoch and functioned as a southern shelter with Yellow River. For this reason, the Korean Peninsula is suggested to be an important geographical region for better understanding phylogenetic relationships and evolutionary histories of B. lenok tsinlingensis across the Northeast Asia. Despite large efforts made to develop several restoration programs in South Korea for B. lenok tsinlingensis, it is still unknown whether these past restoration efforts were successful or fruitless, mainly because of little attention paid to post-restoration monitoring research. Hence, there was a lack of their published official records. In the future, conservation and restoration projects of the Korean lenok populations should consider the genetic data for a better understanding of their ecological and evolutionary trajectories. And finally, we hope that our findings here can help inform on the future effective conservation and restoration plans for B. lenok tsinlingensis populatio ns in South Korea.

Insect growth regulators (IGRs) are compounds that induce developmental anomalies such as premature molting or supernumerary larval stages. IGRs can be divided into 3 groups based on their mode of action, one of which is known as the juvenile hormone agonists (JHAs). Previously, we have developed a novel screening method which can identify juvenile hormone antagonists (JHANs) using a yeast-two hybrid system incorporating Aedes aegypti juvenile hormone receptor complex proteins (MET and FISC). It has been reported that plants may use IGRs in their defenses against insect herbivores. In this study, 171 plant essential oil samples were screened using the yeast-two hybrid system in order to identify JHAs and/or JHANs. Eight out of 171 samples, 4 JHA and 4 JHAN candidates, were selected and their insecticidal activities were investigated against A. aegypti larvae. They showed high levels of insecticidal activities, with LC50 values ranging from 11.2~21.4 ug/ml. These results indicate that plants do use JHAs and/or JHANs as a part of their defenses against insect herbivores. Also, JHAs and JHANs identified in this study could be exploited as effective biological control agents.

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. IGRs are categorized into three types based on their mode of action. One of the them is classified as juvenile hormone agonists (JHAs). Recently, we developed a yeast-two hybrid system that can identify potential juvenile hormone antagonist (JHAN) candidates using Aedes aegypti juvenile hormone receptor complex proteins, known as MET and FISC. In order to identify novel JHAN compounds, approximately 8,700 basic chemical compounds were screened using the yeast-two hybrid system. Among them, 35 compounds were first selected based on consistency of JHAN　activity, and their insecticidal activities were investigated against A. aegypti larvae. These compounds showed insecticidal activities about 1.7~3 times higher than that of pyriproxifen, which is a well known JHA insecticide. These results suggested that these JHANs can be used as effective JHAN insecticides.

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 systems become the targets of two types of IGRs: the Juvenile Hormone Agonists (JHAs) and Juvenile Hormone Antagonists (JHANs). Pyriproxyfen is one of the chemical compounds widely used as JHA to control many kinds of insects while Kanakugiol is a plant-extracted compound which acts as JHAN. The small brown planthopper, Laodelphax striatellus, is one of the most serious pest insects of rice plants because it can transmit the rice stripe virus which often causes significant reduction of yield in the field. In order to analyze the differential gene expressions of L. striatellus upon JHA and JHAN treatment by using next generation sequencing technique, we sprayed Pyriproxyfen and Kanakugiol on 4th instar nymphs of L. striatellus respectively, and extracted total RNA for RNA-seq. The quality-filtered Illumina sequence reads of the control, JHA, and JHAN treated samples were mapped to the reference gene sequences by using the Bowtie2 software. Then the results of mapping by Bowtie2 were analyzed by eXpress software to quantity the differential gene expression.

Recently, Bacillus thuringiensis (Bt) cry genes encoding insecticidal Cry proteins have been widely applied for the construction of transgenic crops resistant to insect pests. This study aimed to construct novel mutant cry1Ac genes for genetically modified crops with enhanced insecticidal activities. Using multi-site directed mutagenesis, 34 mutant cry1Ac genes were synthesized and converted at 24 amino acid residues, located on domain I (8 residues) and domain II (16 residues). These mutant genes were expressed as a fusion protein with polyhedrin using the baculovirus expression system. The expressed proteins were occluded into polyhedra and activated stably to 65 kDa by trypsin. Among these, Mut-N04, N06, and N16 showed high levels of insecticidal activites against larvae of Plutella xylostella, Spodoptera exigua, and Ostrinia furnacalis. Mut-N16, which showed the highest insecticidal activity, is expected to be a desirable cry gene for introduction into transgenic crops. This study could provide useful means to construct mutant cry genes with improved insecticidal activities and expanded host spectrum for transgenic crops.

본 연구는 베트남 하노이 지역에서 8가지 한국 토마토 품종 (핑크탑, 큐피랑, 슈퍼도태랑, 선레드, 선글러브, TP-7 플러스, 러블리 250, 광복)과 1가지 베트남 품종(FM 120)을 대상으로 봄-여름 및 가을-겨울 작기에 따른 수확량 및 과실 특성을 비교 분석하였으며, 그 결과는 다음과 같다. 1. 작기에 따라 초장, 경장, 주당 과실수 및 수확량(kg)은 유의성이 있었으며 가을-겨울 작기에서 높게 나타났다. 전체적으로 한국품종이 베트남 품종에 비해 높은 생육특성을 보인 반면, 과실수와 수확량에서는 베트남 품종이 우수하였다. 2. 토마토 과실의 주요 특성인 과장, 과폭 및 과중은 작기에 따라 유의한 차이를 보였으며 가을-겨울 작기에서 높은 수치를 나타내었지만 TSS 함량은 봄-여름 작기에서 높았다. 건물중과 비타민C 함량은 작기에 따라 유의한 차이가 없었다. 품종별 과실 특성은 한국 품종이 우수하였지만 비타민C 함량은 베트남 품종이 높았다. 3. 봄-여름 작기에서 한국품종의 경우 수확량 및 과실특성에서 가을-겨울 작기 재배에 비해 큰 감소를 보인 반면 베트남 품종은 큰 차이를 보이지 않았다. 따라서 본 실험에 이용된 8가지 한국 품종의 경우 봄-여름 작기보다는 가을-겨울 작기 재배가 적합할 것으로 판단된다.